BCHMLS1

BIOCHEMISTRY
MODULE AND
WORKBOOK
FOR UNIVERSITY OF BAGUIO

STUDENTS’ USE ONLY
COMPILED BY:
ANTONINA C. GUERRERO, RMT, MAED
EVELYN B. SEBASTIAN, RND, MPH
A Self-regulated Learning Module 2

Dear Student,
Welcome to the world of Biochemistry!
This module was created for the use of students enrolled in BCHMLS1
(Biochemistry for the Medical Laboratory Sciences) and BDCHEM1 (Biochemistry for
Doctor of Dental Medicine) in the University of Baguio.
It contains lecture materials designed for individual learning. It also includes simple
laboratory activities, some of which may be performed at home using common household
materials. Most laboratory procedures however, are applications of the lecture and will
make use of reference materials.
This module is divided into 15 submodules designed for lessons on a weekly basis. A
time frame is suggested to finish a certain unit or chapter including laboratory activities.
Make sure to go over the objectives of each lesson to find out what you need to
accomplish at the end of every learning session.
All units contain activities to check your progress as well as formative and
summative assessments. Assignments are also given per unit.
We are hoping that you will find this module helpful in your pursuit of knowledge.
ACG
EBS

TABLE OF CONTENTS
Week Lesson/Activity Page
No. No.
1 Submodule 1: Introduction to Biochemistry 8
Activity No. 1: The cell 21
2 Submodule 2: The Biochemistry of the Cell 23
Activity No. 2: Membrane Physiology: Diffusion 39
Activity No. 3: Membrane Physiology: Osmosis 43
Activity No. 4: Membrane Physiology: dialysis 45
3 Submodule 3: pH and the Chemistry of Respiration 49
Activity No. 5: pH Determination 68
4 Submodule 4: Nucleic Acids and Heredity 71
Activity No. 6: Cell Division and DNA Replication 97
5 Submodule 5: Enzymology 105
Activity No. 7: Enzymes and Digestion 124
6 Submodule 6: The Biochemistry of Digestion 128
Activity No. 8: Saliva 143
Activity No. 9: Bile 146
7 Submodule 7: Introduction to Carbohydrates 150
Activity No. 10: CHO: Physical Properties and General Tests 166
8 Submodule 8: Properties of Carbohydrates 171
Activity No. 11: Qualitative Tests for Specific Carbohydrates 173
Activity No. 12: Reducing Properties of Sugars 186
Activity No. 13: Inversion of Sucrose 190
9 Submodule 9: Metabolism of Carbohydrates 194
Activity No. 14: Starch and Glycogen 219
10 Submodule 10: Metabolism of other Carbohydrates 223
Activity No. 15: Cellulose 239
11 Submodule 11: Amino Acids and Polypeptides 242
Activity No. 16: General Tests for Proteins 251
Activity No. 17: Test for Specific Amino Acids 254
12 Submodule 12: Properties of Proteins 257
Activity No. 18: Properties of Proteins 268
13 Submodule 13: Metabolism of Proteins 271
Activity No. 19: Chromatography of Amino Acids 281
14 Submodule 14: Classification, Properties and Digestion of Lipids 284
Activity No. 20: Physical and Chemical Properties of Lipids 303
15 Submodule 15: Metabolism of Lipids 308
Activity No. 21: Other body Fluids: Urine 321

COURSE CODE: BCHMLS1
COURSE TITLE: BIOCHEMISTRY
COURSE DESCRIPTION:
This course introduces the students to the study of the molecular basis of life. It focuses
on the molecular anatomy of the different biomolecules namely: nucleic acids, carbohydrates,
lipids, proteins and amino acids. It also includes the molecular physiology of the different
biochemical mechanisms involved in the breakdown of complex biopolymers which lead to the
synthesis of important metabolites and the production of energy. Emphasis is also given on the
medical, clinical and human implications from the concepts presented, so designed to enable
students to understand and appreciate the importance of the intricate chemical processes
occurring inside the human body. This course will provide a good foundation for their knowledge
in pursuing their chosen fields of specialization and a stepping stone for pre-medical courses.
REQUIREMENTS OF THE COURSE:
1. Regular Attendance to classes: You must attend online classes and live quizzes
regularly by logging in to our scheduled online activities. Online lectures will be done
through Google meet and/or facebook live. Assessments shall be given through Quizziz,
Pear Deck, Canvas and/or Google forms. For offline students, your attendance will be
monitored through your responses to text information and through timely
correspondence.
2. Submission of required activities: All required activities (assignments, research
work, laboratory illustrations) should be submitted on or before the given deadline.
Deadlines will be posted by the teacher in the google classroom and messenger group
chat. It will also be texted to offline students. For online students, requirements must be
submitted to the teacher’s email address which will be provided during the class
orientation. For offline students, requirements must be submitted via mail or express
courier (e.g. LBC, JRS) addressed to: Instructor’s name, School of Natural Sciences,
University of Baguio, Baguio City.
• Quizzes using google forms will be announced during scheduled lectures.
3. Seventy percent (70%) passing score in all required activities: Quizzes, exams,
assignments, research work, laboratory illustrations.
Computation of grades:
▪ The Course Grade is obtained by combining the lecture and laboratory grades
(50%:50%) for the subject.
▪ Laboratory grade shall be computed as 30% enhancement activities (illustrations;
research work; case study; experiments – when possible) plus 70% class standing
(quizzes and exams).
▪ The cumulative system of computing grades shall be followed. Grades computed for
midterms and finals are considered tentative. The final midterm grade is calculated by
getting 1/3 of the first grading grade plus 2/3 of the tentative midterm grade and the
final grade is computed by getting 1/3 of the midterm grade plus 2/3 of the tentative
final grade.

4. Study/Learning Guidelines:
a. Manage your time properly. As students of higher education (College), you are
expected to be more responsible in paying attention to course schedules,
requirements, and deadlines. Schedule how you will accomplish all the requirements
in all your enrolled courses (reading the modules, reading on research/ enhancement
questions, doing assignments and laboratory illustrations) and focus your attention
when doing your tasks.
b. Observe proper conduct. Despite this online mode of learning, you must still maintain
appropriate behavior at all times. All standards of student conduct outlined in the
University of Baguio Student Handbook remain in full effect during this time of
distance learning. Be honest in answering your quizzes and exams. Work
independently when accomplishing tasks and assignments.
c. Stay motivated. Your future depends on what you do today. Maintain a positive
attitude towards learning and enjoy a fun-learning environment despite the current
circumstances.
d. Maintain a performance of high standard. Give your best in accomplishing all the
assigned tasks. Do not be complacent with just a 70% passing cut-off score.
Remember that this is a board subject, and the best preparation for the
board/licensure examination should be during these formative years. The board
review is but supplementary to the knowledge you have already learned during your
Med Tech education.
e. Communicate properly. Promptly respond to notifications by regularly visiting our
google classroom and messenger group chat. If you have confusions or queries in
any part of this module, I am here to guide you through. Send your academic
concerns using the same online platforms. For offline students, text messages and
mobile calls are welcome during scheduled hours of the day and week. Be guided by
this schedule when communicating:
▪ Respect private hours. I do not always open my laptop/email/messenger 24/7.
Send your queries and/or concerns during regular office hours. For concerns
that need immediate attention, send through mobile text.
▪ Be patient. Messages received between 8 AM to 8 PM will be responded to
within the same day. Messages received after 8 PM will be answered starting 8
AM the next day.
▪ Before calling my mobile number, text first for permission for I might be giving
an online lecture or in a meeting or on private moment at that very instance.
▪ Saturdays and Sundays are for my family and home chores. I shall respond to
queries/messages received during these days within the first office hour of
Monday.
f. Show mutual support. Support one another. Let us all be responsible and supportive
in making this new learning process more effective.
g. Live lecture/Video conferencing guidelines:
g.1 Be punctual. Live lectures/Video conferences will be scheduled during the official
class period/time of this course. Log in to the platform at least 5-10 minutes

before the class period. Prepare your learning materials such as this module,
pens, papers, etc. Attendance will be checked during the lecture/video
conference.
g.2 Maintain professionalism.
- Wear appropriate clothing and set your gadget in an appropriate area. You
may be asked to turn on your video/camera at any time during the lecture.
- Log in using your UB gmail account. Unidentified names like nicknames,
phone models, etc. will not be allowed in the video conference.
- Mute your microphone as soon as you log in to the platform to avoid any
excess background noise. Unmute your microphone when instructed to do so.
- Be courteous. Do not interrupt your teacher or a classmate who is speaking.
You may type your question in the Chat area, or use the “raise hand” feature if
available, and wait until you are allowed to speak.
- Respect privacy. Do not take a screenshot, picture, snapshott, etc. of your
teacher or fellow students, nor make any unnecessary audio or video
recordings.
g.3 Remain focused and engaged. Do not be distracted by your gadget. Keep your
videoconference platform open and do not navigate other tabs or webpages
unless directed by your teacher.
LEARNING COMPETENCIES:
COGNITIVE
(1) Define the different terms encountered in Biochemistry.
(2) Explain the physiological importance of the different bio-molecules.
(3) Explain the chemical reactions, which underlie the results obtained in the
biochemistry laboratory, and relate its importance in the clinical aspect of the course.
(4) Analyze certain basic biochemical processes to explain commonly occurring health-
related problems.
(5) Identify the different parts and the chemical components of a cell.
(6) Define terms associated with CHO;
(7) Describe CHO according to composition, classification and chemical properties;
(8) Discuss the metabolism of CHO in the body;
(9) Explain the mode of actions of different hormones in the maintenance of blood
glucose levels;
(10) Discuss the different processes involved in the maintenance of normal blood
glucose;
(11) Classify proteins according to structure and function;
(12) Discuss the digestion of complex biomolecules in the body;
(13) Discuss the metabolism of amino nitrogen;
(14) Illustrate the structures of the different amino acids;
(15) Describe the metabolism of the different amino acids;
(16) Describe terms associated with lipids;
(17) Describe lipids according to composition, classification, properties;
(18) Discuss the metabolism of lipids in the body;
(19) Describe the nature of enzymes;
(20) Comprehend how enzyme kinetics occur;

(21) Identify the different factors that will influence an enzymatic reaction;
AFFECTIVE
(1) Discern the importance of the knowledge of metabolism in keeping oneself healthy;
(2) Appreciate the significance of the metabolism of different biomolecules in the
maintenance of normal life processes;
(3) Obey protocols in the laboratory for the maintenance of a clean & safe working
environment;
(4) Internalize the relationship that exists among the different metabolic pathways;
(5) Develop a persevering attitude in studying intricate metabolic processes;
PSYCHOMOTOR
(1) Analyze results obtained from experiments to compare with biological concepts;
(2) Sketch an illustration of a typical animal cell;
(3) Perform skillfully the procedures included in the laboratory experiments;
(4) Demonstrate laboratory safety practices and proper waste disposal;
Endorsed by:
Teresa N. Villanueva, RMT, MACT

Dean, School of Natural Sciences

Week no.1
Submodule 1
Introduction to
biochemistry

LESSON: Introduction to Biochemistry
TOPICS:
1. Definition, History and Importance
1.2. Branches of Biochemistry
1.2.1. Molecular Anatomy
1.2.2. Molecular Physiology
1.3. Molecular Anatomy: The Biomolecules
1.3.1. Carbohydrates
1.3.2. Lipids
1.3.3. proteins
1.3.4. Nucleic acids
1.4. Molecular Physiology: Overview on Metabolism 1.4.1. Definition
1.4.2. Functions of Metabolism
1.4.3. Phases of Metabolism
TIME/SCHEDULE: Week 1
LECTURE: 5 Hours
LABORATORY: 4 hours
LEARNING OBJECTIVES:
At the end of the learning session, the student will be able to:
1. Clearly define the different terms encountered in Biochemistry.
2. Explain completely the physiological importance of the different biomolecules.
3. Explain completely the chemical reactions which underlie the results obtained in a
biochemistry laboratory and relate its importance in the clinical aspect of the course.
4. Discern concisely the importance of the knowledge in metabolism in keeping oneself
healthy.
5. Appreciate discerningly the significance of the metabolism of the different biomolecules in
the maintenance of normal life processes

LESSON PROPER
INTRODUCTION:
Organic matters come in different forms. It can be the materials we use in our daily lives
or the foods we consume every day. Nonetheless, these organic substances, primarily those that
come from food, will undergo several processes in the body. This in turn will provide the necessary
materials needed by the body for growth, development, body building, tissue repair, and
regulation. These different processes are included in the study of Biochemistry.
Activity 1. Given the following picture, what can you infer about biochemistry? Write your answer
on the space allotted below.
_____________________________
_____________________________
_____________________________
_____________________________
_____________________________
_____________________________
_____________________________
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_____________________________
_____________________________
Lesson 1. Definition, History and Importance of Biochemistry
Every discovery, words and even meanings has their own history. It may be a result of
experiments, accidental discoveries, or just a word uttered in weird manner. Biochemistry came
into existence due to the efforts of several individuals. Who do you think is the Father of
Biochemistry? Why? It is also necessary for you to be aware on the different terms used in
Biochemistry. This will help you to better understand the processes and to have a general and
uniform understanding of a concept.
Biochemistry is referred to as the study of the molecular basis of life. It is also defined as
the study of biomolecules and the different processes they undergo inside the human body as
they affect the life, health and nutrition status of the individual. A good knowledge in Biochemistry
will help you understand the structure and behavior of biological molecules or biomolecules. This
in turn would allow appreciation of the processes undergone by food once ingested.

Biochemistry is concerned with the physicochemical processes underlying digestion,
absorption, circulation, respiration, metabolism, growth and reproduction. It only shows that
biochemistry has an impact to different processes inside the body. This is primarily due to the fact
that once biomolecules enter the body, they will already be process by some organs and
enzymes. Once processed, these biomolecules will be converted to nutrients and substances
necessary to support body processes such as those mentioned earlier.
History of Biochemistry
1. Freidrich Wohler (1828)
- His experiment laid the foundation for modern biochemistry

- He heated ammonium cyanate (inorganic compound) to form urea (organic compound
found in urine and blood)
- Demonstrated that organic compounds need not necessarily be formed in living organisms
2. Two Major breakthroughs in the history of Biochemistry
a. Discovery of enzymes and its roles

• Enzymes – catalysts of biological reactions
➢ Starch amylase glucose
➢ Amylase from the example is the enzyme necessary to degrade starch
into its simplest form which is monosaccharide. This shows how
significant enzyme is in any biological reactions. With its discovery,
better understanding of metabolic processes was achieved.
• Berzelius – formulated the general principles of catalysts
➢ Lead to the recognition that ptyalin in saliva, pepsin of gastric juice and
amylase of sprouted malt were biological catalysts
• Emil Fischer – studied the catalytic effect of yeast enzymes on a simple
reaction, the hydrolysis of sucrose
➢ Substrate + enzyme = intermediate compound
❖ Proposed the lock-and-key theory of enzyme action (enzyme –
lock; substrate – key). This includes the idea that a particular
substrate is acted upon by a specific enzyme. This will be
discussed further in chapter 5.
❖ Almost all reactions that occur in living cells are catalyzed by
enzymes and thus proceed at very high rates
• Carl Neuberg – introduced the term Biochemistry in 1903 and thus was named
as the father of biochemistry. His notable contribution to science includes the
discovery of the enzyme carboxylase. This is necessary in the decarboxylation
of pyruvic acid. His experience and contribution to the field of biochemistry
paved way to a better understanding of metabolic pathways which was
investigated by later researchers.
b. Identification of Nucleic Acid as information molecules
• Oswald Avery, Colin Macleod and Maclyn McCarty (1944)

➢ They extracted DNA from a toxic strain of Streptococcus pneumonia
and mixed the DNA with a non-toxic strain

• James D. Watson and Francis H.C. Crick (1953)
➢ They deduced the 3D structure of DNA
➢ They discovered DNA replication process, which then transmits
biological information to succeeding generation
• The central Dogma
➢ States that the information encoded in DNA is transcribed to RNA and
then translated to protein
➢ DNA Transcription RNA translation CHON
➢ This is significant in understanding protein synthesis and interrelated to
cell function. A thorough discussion of each step involving central
dogma will be given in Chapter 4.
Activity 2. Answer the following questions and write your answer in the corresponding boxes.
1 2
1. Biochemistry focuses on the
study of these substances and
the processes they undergo in
the body.
2. This was the enzyme
3 discovered by Neuberg.
3. In central dogma, this is
formed after the RNA is
4
translated.
4. In the reaction:
5 Starch amylase glucose
Amylase is the ________.
5. This is the organic compound
formed in the experiment of
Wohler.
Lesson 2. Branches of Biochemistry
Biochemistry, just like any other sciences, has also its different branches. Different
concepts are included in each of its branches. The following lesson will discuss each thoroughly
for better understanding.
Branches of Biochemistry
1. Molecular Anatomy
2. Molecular Physiology
Molecular Anatomy talks about the different structures and properties of the different
biomolecules. Biomolecules are any organic or inorganic matter which when introduced into the
body (eaten, injected or inhaled) will either positively or negatively affect vital life processes. E.g.
food, medicine, chemicals, poisons. They also pertain to carbon-containing compounds that make
up the various parts of the living cell and carry out the chemical reactions that enable it to grow,
maintain and reproduce itself, and use and store and energy. In this case, biomolecules are also

known as biopolymers. They are macromolecules created by joining many smaller organic
molecules, or monomers via condensation.
Carbohydrates are organic compounds containing carbon, hydrogen and oxygen, with a
hydrogen-oxygen ratio of 2:1. These include simple sugars (monosaccharides) and their polymers
(polysaccharides) and contain several hydroxyl groups (polyalcohols). Sugar structures can be
represented in several ways which include:
1. Fischer Projection – linear representation 2. Haworth Projection

- Open chain form i.e. ribose
- Ring form (biochemical form
Proteins are biomolecules that contain an amino group and a carboxylate group as well
as a side chain. These are made up of different amino acids joined by an amide bond or a peptide
bond and its shape is determined by the sequence of its amino acid residues which is encoded
by a gene. The protein function depends on its 3D structure or conformation.
Lipids are generally, water-insoluble organic compounds that form the biological
membrane. The simplest lipids are the fatty acids, long chain hydrocarbons with a carbohydrate
group at one end
Nucleic acids are polymers composed of monomers called nucleotides. Nucleotides

contain a 5-carbon sugar, a heterocyclic nitrogeneous base and at least one phosphate group.
There are two nucleotides, the ribonucleotides and deoxyribonucleotides. In ribonucleotides, the
sugar is ribose while in deoxyribonucleotides, the sugar is deoxyribose. The two nucleic acids are
DNA (deoxyribonucleic acid) and RNA (ribonucleic acid). These will be discussed further in
Chapter 4.
Molecular Physiology is concerned about the functions of the different biological

molecules. This includes the following:
Work within cells

1. Mechanical work – a change of location or posture of an organism, cell or cellular
structure

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2. Osmotic or electrical work – compounds or ions are often moved against a
concentration gradient
Microbenotes.com
3. Synthetic work – a change in chemical bonds required to generate complex molecules

from simple precursors
• The energy required to carry out this work can only come from chemical
bond energy. This is achieved by coupling energetically favorable
reactions to those that require a net energy input
Metabolic pathways are referred to as organized sequence of chemical reactions that are
needed to extract the chemical bond energy from energy supplying compounds and to synthesize
different biological molecules. Important to the understanding of metabolic pathways are the
following terms:
Enzymes – controls the metabolic pathways by catalyzing each of the steps in a pathway
Hormones – intercellular messengers that helps regulate the amount of substrate and
enzymes

Importance of metabolic pathways
- It permits control of the rate and direction of the cellular activity
- It prevents very large chemical bond energy releases which would be damaging to
cells
- It permits branch points
- Allows pathways to be directed (under different circumstances) to different end
products
2 types of metabolic pathways

1. Linear metabolic pathway – series of reactions generates a final product
➢ A B C D
➢ Each reaction is catalyzed by an enzyme
2. Cyclic – series of reactions regenerates the first reactant

➢ A B
D C
Activity 3. ODD-MAN OUT. Given the following combinations, identify which does not belong to
the group.
1. ENZYMES glucagon lactase lipase sucrase

2. CYCLIC METABOLIC PATHWAY Kreb’s cycle Glycolysis Urea Cycle Calvin
cycle
3. HORMONES amylase glucagon insulin ACTH

4. NUCLEOTIDES Ribose Purine Amino Acid Pyrimidine
5. BIOMOLECULES Foods Drugs Nutrients Soap
Lesson 3. Metabolism
METABOLISM is referred to as the sum total of the physical and chemical processes and
reaction taking place among the ions, atoms and molecules of the living body. These processes
are concerned with the role of the cells of an organism to transform energy, maintain their identity
and reproduce.
2 PHASES OF METABOLISM
1. ANABOLISM – is the
constructive phase of
metabolism. It is the process of
synthesis required for the growth
of new cells and the maintenance
of all tissues
➢ combine small molecules
to form complex
molecules
➢ accomplished by
breaking down ATP to
ADP
➢ anabolic reactions often
involve chemical
reductions
➢ the reducing power is
provided by the electron
donor NADPH
➢ anabolism is a divergent
process in which a few
biosynthetic precursors
from a wide variety of
polymeric or complex
products
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2. CATABOLISM – is the destructive phase of metabolism. It is a continuous process concerned

with the production of the energy required for all external and internal physical activity
➢ It involves the maintenance of body temperature and the degradation of complex chemical
units into simpler substances that can be removed as waste products from the body
through the kidneys, intestines, lungs and skin
➢ Functions
i. Serve to capture chemical energy from the degradation of energy-rich fuel
molecules
ii. Allows molecules in the diet to be converted into building blocks
➢ 3 stages of Catabolism
▪ Hydrolysis of complex molecules into component building blocks
• Proteins to amino acids
• Polysaccharides to monosaccharides

• Triglycerides to free fatty acid
▪ Conversion of building blocks to simple intermediates
• Sugars usually enter the glycolysis pathway in the form of glucose
or fructose which are eventually converted to acetyl CoA
• Amino groups are removed from amino acids and the remaining
carbon skeletons enter the catabolic processes at many steps of
glycolysis and the citric acid cycle
• Fatty acids are converted to Acetyl CoA
▪ Oxidation of acetyl CoA and the production of ATP
NB. Anabolic and catabolic processes should be proportionately done in order to maintain the
biological equilibrium or homeostasis in the living body
Catabolic pathway Anabolic pathways
- AKA degradative pathway - AKA synthetic pathway
- Breaks down complex - Form complex end products from
biomolecules to a few simple simple precursors
molecules
Oxidation Reduction Reactions

- Involve electron transfer between molecules
- Dehydrogenase is the enzyme for redox reactions. You will see more of this
enzyme in the different metabolic pathways for biomolecules. Further discussion
regarding this enzyme and reaction will be given in Chapter 5.
- Makes use of cofactors (accepts or donates electrons)
Oxidation
- Involves loss of electrons from a molecule
- Often accompanied by a loss of one or more hydrogen atoms from the molecule
Reduction
- Involves gain of electrons by a molecule
- Often accompanied by a gain of one or more hydrogen atoms by the molecule
ELECTRON ACCEPTORS OR DONORS USED BY DEHYDROGENASE ENZYMES

Oxidized form Reduced form Meaning ATP
(Electron (Electron donor) equivalent
acceptor)
NAD+ NADH Nicotinamide Adenine Dinucleotide (hydride) 2.5
FAD+ FADH Flavine adenine dinucleotide (hydride) 1.5
FMN FMNH Flavin mononucleotide hydride 1.5
FAD – from double to single bond

- Example: succinate to fumarate
NAD – from secondary alcohol to ketone
- Example: pyruvate to lactate
Note of the ATP equivalent since this will be significant in the determination of ATP
produced from a particular metabolic pathway.

Activity 4. Classify the following reaction if anabolic or catabolic.
1. Synthesis of a polysaccharide from monosaccharides ____________________

2. Hydrolysis of pentasaccharide to monosaccharide ____________________
3. Formation of nucleotide from phosphate, sugar and base ____________________
4. Hydrolysis of triacylglycerol ____________________
5. DNA and RNA formation from nucleotides ____________________
Lesson 4. Processes Undergone by Biomolecules
Previously, the different biomolecules were discussed including their functions and
significance. This lesson will give thorough discussion about the different processes undergone
by biomolecules.
Remember that biomolecules are substances such as foods, drugs or nutrients that are
ingested by the body. These include proteins, lipids, and carbohydrates. These different
substances will undergo different processes once introduced to the body.
PROCESSESS UNDERGONE BY BIOMOLECULES

A. DIGESTION
- it refers to the breakdown of large foodstuffs into smaller particles
- 2 forms
➢ Physical digestion – the mechanical conversion of big food into smaller
observable particles
➢ Chemical Digestion – the conversion of big food particles into smaller
absorbable forms with the participation of enzymes, hormones and
digestive juices
B. ABSORPTION
- it refers to the diffusion or movement or nutrients and other ingested materials from
the small intestines (jejunum) into the blood stream
- the process is facilitated by microvilli which are mobile, fingerlike projections that
increases the absorptive area of the GIT
C. ASSIMILATION
- it refers to the selective uptake of specific nutrients by an organ in the body
- in other words, the absorbed nutrients are not uniformly distributed in the body
- For example:
➢ Bones and teeth would take up more of the calcium and phosphorus
➢ Thyroid gland would take up most of iodine
➢ Hair would take up zinc
➢ Bone marrow take most of the iron and copper to make RBC
➢ Adipose tissues would take up most of the fat
➢ Liver would take up most of the sugars
D. UTILIZATION
- the process by which the absorbed nutrients are used by the different cells for a
specific purpose or function
- e.g. ATP (adenosine triphosphate) – energy used by all cells in the body
E. INTEGRATION INTO TISSUES
- the process by which the absorbed nutrients are included or incorporated into the structural
framework of the body like in the bones, muscles, teeth, hair, skin, joints, and ligaments
F. BIOTRANSFORMATION AND METABOLIC DEGRADATION

-
the process by which all harmful and potentially toxic materials introduced into the
body (like food, preservatives, food coloring, and chemicals like formalin) are
inactivated or detoxified by the liver into something non-toxic or even less toxic
thus, no significant harm is done on the body
- 2 forms
➢ Metabolic degradation – breakdown of complex substances into simpler
ones that act as active metabolites or end products for energy production
➢ Biosynthesis – combination of simpler substances to build complex
substances for cell repair, growth and reproduction
G. EXCRETION
- process by which metabolic wastes are finally expelled or removed from the body
- these wastes when allowed to accumulate inside will destroy cells and tissues so
these must be dispensed off fast
- Organs of excretion:
➢ Kidneys – urine
➢ Lungs – volatile acids in the form of CO2
➢ Skin – sweat (hypotonic NaCl)
➢ GIT – stool or feces or undigested residue of food (excreted through
defecation
Activity 4. Trace the processes undergone by cheeseburger (with consideration of the different
biomolecules present in the given food). Identify the organs/cell part/tissue involved in the different
processes (if applicable).
Activity 4
Needs
Unsatisfactory Satisfactory Outstanding
Improvement
3 pts 7 pts 10 pts
5 pts
Content & -Content is - Content is - Content is - Content is
Development incomplete. somewhat accurate comprehensive and
- Major points are incomplete - Organs/cell accurate.
not clear. - Major points are part/tissue is/are not - Correct organs/cell
-Specific examples addressed, but not identified properly part/tissue is/are
are not used. well supported. - Content is clear. identified
-Specific examples - Content is clear.
do not support topic. -Specific examples
are used.
Organization & - Organization and - Structure/flow of - Structure/flow is -Structure/flow of
Structure structure detract the paper is not mostly clear and the paper is clear
from the concept. easy to follow. easy to follow. and easy to follow.
- The process in - Transitions need - Transitions are - Transitions are
incorrect. improvement. present. logical and maintain
- Most words were - The process is in - The process is in the flow of thought
misspelled. different order. different order. throughout the
- There are 3-5 - There are 1-2 paper.
words that are words that are - The process is in
misspelled. misspelled. correct order.
- Spelling is correct.

ASSESSMENT
A. IDENTIFICATION. Given the following descriptions/ definitions, identify what is being

described/ defined.
_______________________________ 1. This a biomolecule that contains the elements

carbon, hydrogen, nitrogen, and oxygen.
_______________________________ 2. The biological process involved when thyroxin
uptake iodine.
_______________________________ 3. In the following reaction:

Lactose lactase glucose + galactose
What is the enzyme?
_______________________________ 4. Who is the proponent of the lock and key
theory?
_______________________________ 5. What organ is responsible in
biotransformation?
_______________________________ 6. The formation of polypeptide from amino
acids shows what reaction?
_______________________________ 7. What form of digestion is shown when an
enzyme is required?
_______________________________ 8. How many ATP is produced by the reduction
of NAD?
_______________________________ 9. Enumerate one importance of the metabolic
pathway.
_______________________________ 10. When fatty acid is converted to acetyl CoA,
what reaction is shown?

LABORATORY
INTRODUCTION:
Underlying each and every biological function is a chemical reaction. The cell is the basic
structural unit of all living things. All processes pertaining on how life is sustained depends on
how the cell harnesses the food that we eat and converts it to energy and building blocks used
for growth and reproduction.
The cell is made up of different biomolecules. Four of these biomolecules – proteins, nucleic
acids, carbohydrates and lipids – are among the most complicated substances known. All cells
are surrounded by a cell membrane, which is the “skin” or envelope containing the cell contents.
The contents of the cell minus the nucleus are the cytoplasm, which can be divided into the cytosol
and the organelles.
Name: _____________________________ Date: _____________________________

Date Assigned: _____________________ Date submitted: _____________________
Activity No. 1
THE CELL
OBJECTIVES:
At the end of the laboratory session, you should be able to

1. enumerate and describe correctly the different parts of the cell
2. relate completely the importance of the cell to biochemistry
3. illustrate the cell and label its parts accurately
MATERIALS:
Pencil
Textbooks Coloring materials
PROCEDURES AND OBSERVATIONS:
Scoring rubric
5 points are given for every illustration made
o 5 – outstanding – the illustration shows genuine effort on the part of the maker and all parts are
labelled with no misspelled words
o 4 – excellent – the illustration shows above average effort on the part of the maker, 1 or 2 parts are
not labelled and with misspelled words
o 3 – Good – the illustration shows average effort on the part of the maker, 3 to 4 parts are not labelled
and with misspelled words
o 2 – Fair – the illustration shows below average effort on the part of the maker, 5 or more parts are
not labelled and with misspelled words
o 1 – Poor – the illustration shows little effort on the part of the maker, and the illustration is not
labelled
1. Diagrammatic illustrations
a. Look for an illustration of the following and label their parts with ink.
➢A typical animal cell
➢The fluid mosaic model of the cell membrane
➢The mitochondria

QUESTIONS FOR RESEARCH
Scoring Rubric
10 points for the whole research paper
o 10 – outstanding – no mistakes identified
o 8 – excellent – 2 - 3 mistakes or missing concepts identified
o 6 – Good – 4 - 5 mistakes or missing concepts identified
o 4 – Fair – 6 -7 mistakes or missing concepts identified
o 2 – Poor – 8 or more mistakes or missing concepts identified
o 0 – No research paper was submitted
1. In a tabulated form, enumerate the parts of the cell, a brief description and its function.
2. Why is the mitochondrion called the “powerhouse of the cell”?
3. Why are lysosomes called the “suicide bag of the cell”?
4. Why are ribosomes called the “protein factories of the cell”?
5. Name an organ that is expected to have numerous Golgi bodies. Why?
6. Why do muscle cells contain numerous mitochondria?
7. What is the importance of this activity in relation to biochemistry?

WEEK NO 2
SUBMODULE NO. 2
THE CELL

LESSON: THE CELL
TOPICS:
2.1. Definition, History and the Cell Theory
2.2. Basic classification of Cells
2.2.1. Prokaryotes vs. Eukaryotes
2.2.2. Somatic cells vs. Sex cells
2.2.3. Animal cells vs. Plant cells
2.3. Chemical Composition and Functions of the Cell and its Organelles
2.3.1. Nucleus
2.3.2. Cytoplasm
2.3.3. Cell Membrane and membrane physiology
2.3.4. Mitochondrion
2.3.5. Endoplasmic Reticulum
2.3.6. Golgi Apparatus
2.3.7. Ribosomes
2.3.8. Lysosomes
2.3.9. Peroxisomes
2.3.10. Cytoskeleton
TIME/SCHEDULE: Week No. 2
LECTURE: 4 HOURS
LABORATORY: 5 HOURS
At the end of the learning session, the students will be able to:
1. Define clearly the different terms encountered in Biochemistry in relation to cell.
2. Identify correctly the different parts and the chemical components of a cell.
3. Clearly explain the physiological importance of the different bio-molecules.
4. Sketch an illustration of a typical animal cell.

LESSON PROPER
INTRODUCTION
The cell is the basic unit of life. Before a system is even formed, there is a need for cells
since cells form tissues, tissues to organs, and organs to systems.
Activity 1. Given the following illustration of a cell, label the different parts. Misspelled words and
any form of imposition will make the answer wrong. (10 points)
1
2
10
3
9
4
5
Lesson 1. Cell and its Functions and the Nucleus
8 CELL is an independent, simplest structural unit of life. It is the fundamental unit of

biological activity and is capable of reproduction. Cells come in a variety of shapes and sizes. 6
Cellular diversity permits organization of cells into more complex tissues and organs. 7The average
adult has nearly 100 trillion cells and there are about 200 different types of cells.
The cell performs several functions which include the following:
1. Nutrition
- the process by which cell obtain food molecules to support their other activities
2. Digestion
- the process by which food particles are broken down into smaller, soluble units suitable
for cell use
3. Absorption
- the process by which the cell absorbs form their environment
- examples are water, minerals and other materials essential for life
4. Biosynthesis
- the cells organize complex chemicals from building units or substances

5. Excretion
- by products of cell activities which are not needed for further cell functioning of the body
6. Egestion
- soluble, non-digested particles are eliminated by the cell
7. Secretion
- substances that are synthesized by the cells are expelled from the membrane
- the elimination influences extracellular activities and helps in the functioning of the body
- examples are hormones
8. Movement
- includes locomotion of cells by means of special structures like cilia and flagella
9. Irritability
- Cells react to external factors or conditions around them
- Cells may also alter their functions in response to this external factor
10. Respiration
- breaking down food molecules into chemical energy
11. Reproduction
- the cell copies or replicates it DNA and increase its number by cell division
- cells give rise to new cells
TYPES OF CELLS
A. PROKARYOTIC CELLS B. EUKARYOTIC CELLS
-- primitive types with not distinct nucleus and - more developed cells because they have definite
cytoplasm nuclear and cytoplasmic structures
- e.g. human cells (about 75 trillion
Recall the activity you did for the laboratory. Remember that you were required to look for
an illustration of an animal cell. Further, you were also given the task of discussing the roles of
the different organelles of the cells. The previous activity will help you understand and relate the
answers to the following discussion.
The cell comprises of organelles. These are specialized cellular parts that have specific
function and is considered analogous to organs.
ORGANELLES OF LIVING CELLS

1. NUCLEUS
- master control of the cell
- it directs, orders and regulates all the metabolic activities of the cell
- an oval, round or elliptical structure usually centrally located inside the cell
- it has a double layered membrane
General Rule:
- there is 1 nucleus in one cell except in erythrocytes (RBC’s) and Thrombocytes
(platelets) which are non-nucleated
- on the other hand, muscle cells are multinucleated

Abnormalities in the nucleus
- abnormalities in the nucleus which may suggest cancer or malignant degeneration
➢ a very big nucleus, out of proportion to the cytoplasm
➢ multiple nuclei
STRUCTURES FOUND IN THE NUCLEUS
a. NUCLEOLUS
- small, discrete, round, densely staining structure
- made up of RNA
- the more nucleoli, the faster in multiplying and dividing
➢ intensely staining concentration of RNA
➢ it is known to produce most of the RNA especially rRNA which gives rise
to ribosomes
b. CHROMOSOME
- linear strands of chromatin material which contain the genes which represent all
the traits of an individual
- the genes are composed of segmented DNA
c. Nuclear Envelope
- consists of 2 membranes
- outer membrane is porous – serves as passageway of information
- outer envelope is continuous with the endoplasmic reticulum
- The outer envelope also performs some secretory and transport processes
- The inner membrane maintains stable relationships with the genetic material
- Pores – considered as gateways of exchange of information from the nucleus to
the cytoplasm
- The nuclear envelop encloses the karyoplasm which in turn is suspended with
DNA and RNA
- DNA – copy information form genetic material (Transcription)
- RNA – Genetic information is used in protein synthesis (translation)
- The nuclear envelop governs nucleocytoplasmic interactions

Have you ever wondered why you have curly hair when most of your family members have
straight hair ? Or you are fair whereas, they are not ? Do you often hear people say you got your
father’s character or your mother’s dimples ? These are all part of hereditary traits. Read through
the next topic to understand the concept better.
TYPES OF TRAITS
1. DOMINANT TRAIT
- it is one which is present or evident or manifested in majority of the offspring or
children in every generation
- e.g. black hair is dominant over blond hair
2. RECESSIVE TRAIT
- is one which may be seen only in a minority of offspring’s
- the trait may even disappear in one generation but will re-appear in succeeding
generations
TYPES OF EXPRESSION OF HEREDITARY TRAITS

1. PHENOTYPES
- the physical observable aspects of heredity as handed down from parents to
offspring’s
- e.g. color of eyes, bridge of the nose, dimples
2. GENOTYPE
- the non-observable, non-physical aspects of heredity
- e.g. IQ, talent, allergy (asthma), diabetes, epilepsy, hemophilia, insanity, color
blindness
Activity 2. Given the following descriptions, identify what organelle is being described.
___________________________ 1. This is the powerhouse of the cell.

__________________________ 2. This is the site of protein synthesis.
__________________________ 3. Which endoplasmic reticulum contains the
ribosomes
__________________________ 4. This is the primary site for packaging cellular
secretions.
__________________________ 5. This is known as the suicide bag of the cell.
Lesson 2. The Cytoplasmic Organelles
2. CYTOPLASMIC ORGANELLES
- living structures which actively participate in the metabolism in cells
- these are “little organs” that serve a specialized function
A. RIBOSOMES

- “factory sites” for the manufacture
of cell protein
- Structures which contain RNA
- These are the sites where amino
acids are joined together by
peptide bonds to form a
polypeptide chain forming a new
protein
B. GOLGI APPARATUS
- a specialized portion of ER
- the primary site for packaging
cellular secretion
- Site of synthesis of large
carbohydrates
- Serve as temporary storage
depots for cellular secretions
C. LYSOSOMES
- known as the “suicide bag of the cell”
- encapsulated granules which contain a very strong proteolytic enzyme (e.g.
hyaluronidase, acid hydrolase)
Note: when microbes or toxins enter the cell, these enzymes are released in order to destroy or
inactivate the invading microbes. Therefore, lysosomes are for intracellular defense mechanism
D. MITOCHONDRIA
- known as the “powerhouse of the cell”
- an ovoid or elliptical structure having 2 layers: the outer layer is continuous while
the inner layer has infoldings or invaginations called cristae matrix
- the only organelle capable of generating energy in the form of ATP
- the only organelles which contains DNA (deoxyribonucleic acid) used for self-
replication or the capacity to reproduce copies of its own

Functions:
• They are the center of cellular
respiration
• They are found in cells that
require substantial amounts of
ATP like the muscle cell
• They are also found in the neck
of the spermatozoa
• Synthesis of ATP from citric acid
• Assist on steroid biosynthesis
• Fatty acid oxidation
• Nucleic acid synthesis
E. ENDOPLASMIC RETICULUM
-composed of network of tubes,
tubules, and microtubules
connecting the nuclear membrane
and cell membrane
- this system of tubules act as
excretory, respiratory and
circulatory passageway of
substances in and out of the cell
such as O2, CO2 and cellular waste
products
-
Types of Endoplasmic Reticulum
A. Rough Endoplasmic reticulum – contain ribosomes
- the ribosomes are connected to the membrane by a ribosome-binding
protein celled ribophorins
- believed as passageway of proteins manufactured by the ribosome
- used as a means of communication
- they channel products from the outside and other parts of the cell
- thought as the cell’s membrane factory
- found abundantly in the pancreas
B. Smooth Endoplasmic reticulum – do not contain ribosomes

▪ metabolizes small proteins
▪ contains cellular detoxification mechanisms
▪ seems important in synthesizing and secreting steroid hormones,
enzymes of protein metabolism and enzymes of lipid synthesis
▪ functions in cholesterol synthesis and breakdown, fat metabolism
and detoxification of drugs
3. CYTOPLASMIC INCLUSIONS
- these are inert bodies which have no effect on cellular activities
A. VACUOLES
- spaces within the cytoplasm which serve as:
➢ temporary dumping site of cellular garbage

➢ storage of glycogen
➢ storage of fat
B. PIGMENTS
2 types according to origin
1. Exogenous origin
- picked-up from the outside of the body
e.g. Dust in the lung, Minerals in bones, lipochromes and
carotenoids form food
2. Endogenous in origin
Hemoglobin- gives the red pigmentation of blood as has several
metabolites that could give rise to endogenous pigments seen in cells
Hemosiderin – metabolite of hemoglobin
- golden brown pigment seen in phagosomes in the liver and the spleen
Bilirubin – metabolite of hemoglobin
- a yellowish pigment that is non-iron containing found in the liver cells
Melanin – a brown-black pigment found in the skin and eye
Lipofuchsin – a brownish pigment seen in the heart, liver, CNS as
the animal ages
- may be found in the nucleus and the cytoplasm
- the origin and function of lipofuchsin is unknown
C. FAT DROPLETS
- found as small globules within many cells
- most prominent in adipose tissues
- they are stored as triglycerides (liquid at body temperature
D. CRYSTALS
- usually found along steroid-secreting cells
- found in the nucleus and cytoplasm
- origin and function is also unknown
E. GLYCOGEN
- found in the liver and skeletal muscles
- found in small clusters called rosettes
- an individual glycogen measures 15-30 nm in diameter
- it is associated with the smooth endoplasmic reticulum within lysosomal particles
Lesson 3. The Cell Membrane
4. CELL MEMBRANE
- also known as plasmalemma or plasma membrane
- it is the analogue of cell wall in plants

Layers of Cell Membrane
A. Mucopolysaccharide or carbohydrate layer – strongest layer of the cell membrane
B. Outer protein layer
Peripheral protein - attached to the inner or outer surface of the membrane, do not extend
through it
C. Double layered lipid coat or lipid bilayer – made up of phospholipids, cholesterol and
glycolipids. This is the most important layer for selective permeability. The lipid bilayer is
permeable to oxygen, carbon dioxide, water and steroids, but impermeable to glucose
D. Inner protein layer - extend into or through the lipid bilayer
Transmembrane proteins - span the entire lipid bilayer. These act as channels and
transporters to assist the entrance of certain substances, for example, glucose and ions
Functions of the Cell Membrane

A. Protection
- protects inside of the cell from external forces (microbes, toxins and chemicals)
B. Limits
- it limits territorial boundaries of the cell
C. Morphology
- give size and shape to the cell
D. Selective permeability
- does not allow some substances to pass thru and allows the passage of only
specific substance
Factors Affecting Passage of Substances

1. Degree of ionization (non-ionized form, more diffusible)
2. Lipid solubility
3. Water solubility
4. Size of the substance
Membrane transport
Activity 3. Answer the following:
1. Define:
a. Solution _____________________________________________________________
______________________________________________________________________
b. Solute ______________________________________________________________

c. Solvent _____________________________________________________________
2. What are the different chemical components of the cytoplasm?

____________________________________________________________________________
____________________________________________________________________________
2. Discuss the role of each components of the cytoplasm into the cell.
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________.
Classification of fluids in the body

Intracellular fluid
- Nucleoplasm + cytosol
- A solution containing small amounts of gases (O2 and CO2), nutrients and salts
Interstitial Fluid
- The fluid that continuously bathes the exterior of the cells
- Contains nutrients (amino acids, sugars, fatty acids, vitamins), regulatory substances
(hormones, neurotransmitters) salts and waste products
Selective permeability
- A barrier allows some substances to pass through it while excluding others
- The plasma membrane is said to be selectively permeable to substances
This particular concept is further explained and observed with the performance of activity
number 2 which is placed at the end of the chapter.
2 types of movements in the cell membrane

Passive (transport) processes Active (transport) processes
- substances move across cell membranes - cell uses energy, primarily from the
without the input of any energy; use the breakdown of ATP, to move a substance
kinetic energy of individual molecules or across the membrane
ions - Requires specific carrier CHONS
- No need for carrier CHONS (proteins) as - Even against a concentration gradient
well as energy or ATP
- Osmosis
Facultative or Facilitated Transport Active Transport

- Does not require energy - cell uses energy
- Needs specific carrier proteins - Requires specific carrier CHONS
- Cannot move substances against - Even against a concentration gradient
concentration gradient

Try to look at the pictures closely. Previously, a comparison between active and passive
processes was given. It was clearly stated that the main difference between the two is the need
for carrier protein and energy of active process whereas, passive requires neither. Note however,
that the transport mechanism that requires just the carrier protein is known as facilitated or
facultative transport. Active transport requires both energy in the form of ATP and carrier protein.
Two types of passive transport processes

Diffusion
- The process by which molecules (and ions) tend to scatter themselves throughout the
available space
- Molecules move down their concentration gradient
- Factors affecting the speed of diffusion
o Size of molecules
o Temperature
o Presence of other molecules
o Distance/size of container
Molecules will move passively through the plasma membrane if:
- They are small enough to pass through its pores
- They can dissolve in the fatty portion of the membrane
Simple diffusion
- Unassisted diffusion of solutes through the plasma membrane
o Lipid soluble: fats, fat-soluble vitamins, oxygen, carbon dioxide
o Small molecules: ions such as chloride
Facilitated diffusion
- A protein “carrier” is needed as a transport vehicle
- It provides a means for certain substances (glucose), that are both lipid-insoluble and too
large to pass through the membrane pores
- No expenditure of energy because they follow the concentration gradient
Importance of diffusion
- It saves the cell a great deal of energy
- Glucose and oxygen continuously move into the cell (where they are in lower
concentration)
- Carbon dioxide continually moves out of the cell into the blood (where it is in lower
concentration)
These particular concepts can be well-understood with the performance of Activity 2 which
can be found at the end of the chapter. You will perform some procedures for you to appreciate
and observe how these different processes take place as well as the factors affecting each.

Osmosis
- Diffusion of water through a selectively permeable membrane
o Because water is highly polar, it is repelled by the lipid core of the plasma
membrane
Figure 1. Filtration Figure 2. Osmosis
- Osmotic pressure
o The tendency of a solution to hold water or pull water into it
o The higher the solute concentration, the greater the osmotic pressure
- Tonicity
o the ability of a solution to change the size and shape of cells by altering the amount
of water they contain
Type of solutions
Isotonic
- Solutions that have the same solute and water concentration as cells do
- Cause no visible changes in cells
- When infused into the bloodstream, the red blood cells retain their normal size and disk-
like shape
- Examples: Ringer’s lactate, 5% dextrose, 0.9% saline solution
Hypertonic solution
- A solution that contains more solutes, or dissolved substances, than there are inside the
cells
- Water is in greater concentration inside the cell than outside and thus it follows its
concentration gradient and leaves the cell
- Crenation – cell shrinking
- It is given to patients with edema
Hypotonic solution
- A solution that contains fewer solutes than the cell does
- Water rushes into the cell as it follows its concentration gradient
- The cell will swell and then bursts
- Cytolysis – cell bursting
- Examples: distilled water, tea, cola, apple juice and sports drinks
- Given intravenously to extremely dehydrated patients (but with care)

Tonicity and its effect to RBCs
Filtration
- The process by which water and solutes are forced through a membrane (or capillary wall)
by fluid, or hydrostatic pressure
- Pressure gradient – gradient involved in filtration
o It pushes solute-containing fluid (filtrate) from the higher-pressure area to the
lower-pressure area
- Occurs in the kidneys
Sodium-potassium pump
- Simultaneously carries sodium ions out of and potassium into the cell
- Needed for normal transmission of nerve impulses
- Na+ - extracellular cation
- K+ - intracellular cation
- “No pump-no transport”
Bulk transport
Two types
- Exocytosis
- Endocytosis
Exocytosis
- Moves substances out of the cells
- A means by which cell actively secrete hormones, mucus and other cell products or eject
certain cellular wastes
- The product to be secreted is packaged first in the golgi body
Endocytosis
- An ATP-requiring processes that take-up, or engulf, extracellular substances by enclosing
them in a small membranous vesicle
- Types
o Phagocytosis – cell eating
▪ Happens when cell form pseudopods (flowing cytoplasmic extension)
o Bulk-phase endocytosis (Pinocytosis) – cell drinking
▪ Happens when the plasma membrane invaginates to form pits and then its
edges fuse around the droplet
▪ Routine function (cells in the lining of SI and kidney tubule cells)

Assessment:
Given below is the picture of the cell, you identify/write what is asked for in the spaces
provided after the picture. Erasures and wrongly spelled words will be considered as
wrong.
18. What is its strongest layer?
19. Function
16. What is synthesized here?
20. Name
17. How much % of RNA is present?
7. Name
5. Name
6. True or False. It is the only organelle that
8. True or false. It is
contains DNA.
a specialized
portion of ER
consists of 3-20 3. Where is this found?
flattened, 4. What is manufactured and transported in
membranous sacs this organelle?
called cisternae.
9. What is being
synthesized by
this?
14. Name
10. True or False. It functions in cholesterol 15. What is being produced here?
synthesis and breakdown, fat metabolism and
detoxification of metals. 1. True or False. This is where most chemical reactions occur.
11. True or False. It metabolizes calcium. 2. Which of its different compositions has no storage form?
12. Name
13. Which of its structure separates the genetic
material from the cytoplasm?
1. _______________________________________________________
2. _______________________________________________________
3. _______________________________________________________
4. _______________________________________________________
5. _______________________________________________________
6. _______________________________________________________
7. _______________________________________________________
8. _______________________________________________________
9. _______________________________________________________

10. _______________________________________________________
11. _______________________________________________________
12. _______________________________________________________
13. _______________________________________________________
14. _______________________________________________________
15. _______________________________________________________
16. _______________________________________________________
17. _______________________________________________________
18. _______________________________________________________
19. _______________________________________________________
20. _______________________________________________________
MORSE TYPE: Match each group of five (5) numbered items to two (2) letter choices.
Choose your answers from the given choices.
A = if the numbered item is related to item A
B = if it is related to Item B
C = if it is related to both A & B
D = if it is related to neither A nor B
_____21. Passive transport A. Facilitated diffusion

_____22. Active process B. Simple diffusion
_____23. Follow law of osmosis
_____24. Need for specific carrier protein
_____25. Movement against a concentration gradient
_____26. Osmolality = Intracellular fluid A. Hypertonic solution

_____27. Osmosis of water from ICF to ECF B. Isotonic solution
_____28. Causes water to move into the cell
_____29. Distilled water
_____30. Given to dehydrated patient

LABORATORY:
INTRODUCTION:
The plasma membrane of the cell is said to be selectively permeable, allowing water to
freely pass through while regulating the movement of solutes. The intracellular fluid and the
extracellular environment of the cell are aqueous solutions primarily composed of water and a
variety of dissolved solutes such as sugars, amino acids and ions. The difference in the
concentration of the solvent and solutes in and out of the cell determines their movement across
the cell membrane. In this activity, you will be able to demonstrate some of the basic principles
of membrane physiology.
All molecules and ions in the body fluids, including water molecules, are in constant
motion, each particle moving its own separate way. This continual movement of molecules among
each other in liquids or in gases is called diffusion. The rapidity with which molecule diffuses from
one point to another is less the greater is the molecular size, because large particles are not
impelled so intensely by collisions with other molecules
Name: _____________________________ Date: _____________________________

ACTIVITY NO. 2
MEMBRANE PHYSIOLOGY: FACTORS AFFECTING DIFFUSION
OBJECTIVES

1. understand correctly the principles involved in diffusion
2. demonstrate diffusion confidently using simple procedures
3. appreciate completely the importance of the homeostasis of solutes and solvents in and
out of the cell
MATERIALS:
Food coloring (any color) distilled water

4 same size drinking glass pencil
Ruler coloring materials
Sucrose/Table sugar
PROCEDURES AND OBSERVATIONS
Scoring rubric
For Procedures:
- 3 points is given for every short procedure done that requires an immediate written observation
o 2 points for doing the procedure correctly
o 1 point for the answer
- 1 point is given for items that do not require performing a procedure
For Rationalization
- 3 points is given for every rationalization made
o 3 – all concepts were mentioned and a clear rationalization was made
o 2 – one or 2 concepts were not mentioned and the rationalization is not clear

o 1 – 3 or more concepts were not mentioned and the rationalization is not clear
o 0 – no rationalization was made
For hypothesis/Generalization/Conclusions
- 3 points is given for every generalization or hypothesis made
o 3 – all concepts were mentioned and a clear hypothesis, generalization or conclusion was derived
from the activity
5 points are given for every graph made
o 5 – outstanding – the graph shows genuine effort on the part of the maker and all parts are labelled
with no misspelled words
o 4 – excellent – the graph shows above average effort on the part of the maker, 1 or 2 parts are not
labelled and with misspelled words
o 3 – Good – the graph shows average effort on the part of the maker, 3 to 4 parts are not labelled
o 2 – Fair – the graph shows below average effort on the part of the maker, 5 or more parts are not
o 1 – Poor – the graph shows little effort on the part of the maker, and the illustration is not labelled
DIFFUSION
Diffusion is defined as the movement of molecules from a region of high concentration to a low
concentration without the assistance of a transport protein. Molecules are propelled by kinetic
energy (the energy of motion). The botanist, Robert Brown, was the first person to observe the
random movement of small particles which is now defined as the Brownian movement.
Several factors can influence the rate of diffusion. These include the steepness of the
concentration gradient, temperature, and size of the molecule. Molecules will move by diffusion
until they reach a state of dynamic equilibrium, equal movement of molecules in both directions.
1. Diffusion through agar gel: Effect of Molecular weight

Agar is a carbohydrate derived from seaweed that will form a gel-like solid when mixed with water,
heated and cooled. Materials can diffuse through agar easily because there are water channels
with the matrix of carbohydrates. Should you use methylene blue and methyl red as materials to
determine the effect of molecular weight in diffusion though an agar gel, which between the two
will diffuse faster? Why?
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________.
2. Diffusion: Effect of Temperature

a. Put ¾ full of hot water in a drinking glass.
b. In another drinking glass, put ¾ full of cold (not room temperature) water.
c. Place in a location where they can remain undisturbed for at least two minutes
d. Carefully place one small drop of food coloring (same color) on the surface of the
water in the center of each beaker. Care should be taken not to touch the drinking
glass or disturb the water when adding the food coloring. The water should be as
still as possible.
e. Observe the pattern of diffusion from the side.
f. Does the drinking glass differ?
________________________________________________________________
Describe what you see.

________________________________________________________________
________________________________________________________________
________________________________________________________________
g. Estimate the percent of water in the glass that is covered every 30 seconds for 5
minutes. Initially (time = 0 sec.) there will be 0% colored. Round your answers to
the nearest 10 percent (10%, 20%, 30%, etc.).
Time Rate of Diffusion in %

Hot water Cold water
0s
30 s
1 min
1.5 min
2 min
2.5 min
3 min
3.5 min
4 min
4.5 min
5 min
h. Prepare a graph using a graphing paper showing the effect of temperature on the
rate of diffusion. Label and place the appropriate legends in your graph. Pass this
together with the questions for research.
i. At which temperature did diffusion occur the fastest? ______________________
j. What is the effect of temperature on the rate of diffusion?
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
Explain your answer.
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
k. Document your result and place it together in your questions for research. Label
properly.
3. Diffusion: Effect of other Substances

a. Place ½ full of water into two drinking glass.
b. Dissolve completely 2 g (1/4 teaspoon) of sucrose or table sugar in one of the
drinking glasses.
c. Let the beakers stand for at least a minute or so undisturbed.
d. Place 1 drop of food coloring or ink (same color) simultaneously into the two
beakers being careful not to disturb it.
e. Let stand for five minutes
f. In which of the beakers did diffusion occur faster? ______________________
g. What is the effect of the presence of other substances on the rate of diffusion?
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________

Why is this so?
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
h. Document your result and place it together with your questions for research, label
it properly.
CLEAN-UP SUGGESTIONS
- The beakers/drinking glass containing the food coloring could be safely poured down the
drain.
QUESTIONS FOR RESEARCH:
Scoring Rubric
1. Describe how the chemical composition of the cell membrane regulates the entry and exit
of solute and solvent into and out of the cell.
2. Differentiate the following
a. Facilitated diffusion via carrier proteins and facilitated diffusion via ion channels
b. Primary active transport and secondary active transport
c. Endocytosis, exocytosis and transcytosis

LABORATORY:
INTRODUCTION:
Red blood cells consist of an envelope and meshwork called the stroma which encloses
a solution of hemoglobin and various salts. The envelope behaves as a semi –permeable
membrane allowing the solvent to enter or leave the red blood cell but not the solutes.
If the RBC are placed in a hypertonic solution, water passes out from the RBC and
cause them to shrink or crenate. If RBC is placed in a hypotonic solution, water enters the cell
and causes them to swell and maybe even burst. In isotonic solutions, the flow of water into or
from the cell is equal and no effect on the cell is observed.
Name: _____________________________ Date: _____________________________

ACTIVITY NO. 3
MEMBRANE PHYSIOLOGY: OSMOSIS
OBJECTIVES

1. understand correctly the principle involved in osmosis
2. appreciate completely the importance of the homeostasis of solutes and solvents in and
out of the cell
MATERIALS
reference material pen/pencil
Scoring rubric
5 points are given for every illustration made
o 4 – excellent – the illustration shows above average effort on the part of the maker, 1 or 2 parts
are not labelled and with misspelled words
o 3 – Good – the illustration shows average effort on the part of the maker, 3 to 4 parts are not
o 2 – Fair – the illustration shows below average effort on the part of the maker, 5 or more parts
labelled
1. Download a picture of RBC added with solutions in different concentrations as indicated below.
Label the picture properly and describe what you see.
Distilled water

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
0.9% NaCl
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
10% NaCl
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
5% glucose
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
Which of the solutions used is hypotonic? __________________________________
Which of the solutions used in hypertonic? _________________________________
Which of the solutions used in isotonic? ____________________________________
Which of the solutions is isosmotic? _______________________________________
Scoring Rubric
1. What is osmosis? State its importance

2. Differentiate between semi-permeable and selectively permeable with regards to the
function of the cell membrane
3. What is tonicity? How does this affect the function of the cell?
4. Differentiate the following
a. Isosmotic from isotonic solution
b. Hypertonic from hypotonic solution
5. Are isotonic solutions also isosmotic? Explain your answer and give examples.
6. What is D-5-W? What type of solution is this and what is its importance?

LABORATORY
INTRODUCTION
Animal membranes, cellophane and collodion have pores with diameters of 7 angstrom.
Molecules with diameters of less than 7 angstrom can therefore easily pass through the pores of
these substances; but larger molecules like colloids are retained in the semi–permeable materials.
This is the basic principle of dialysis.
Colloids are aggregates of molecules with diameters ranging from 1micron (1 micrometer) to 100
microns (100 µm). These large molecules remain suspended in the solvent indefinitely. Such a
system is known as a colloidal dispersion or colloidal solution. The suspension of tiny particles of
one substance are called the dispersed phase and the medium where the particles are dispersed
is called the dispersion medium. The colloidal particles are unaffected by gravity. There are two
types of colloidal dispersion, the emulsoids and the suspensoids.
The protoplasm that makes up our cells is a complex colloid that comprises a dispersed phase of
proteins, fats, and other complex molecules in a continuous aqueous phase.
Name: _____________________________ Date: _____________________________

ACTIVITY NO. 4
DIALYSIS
OBJECTIVES

1. understand fully the basic principles of dialysis using simple procedures
2. appreciate totally how dialysis occurs in the body
3. prepare correctly a simple set-up to demonstrate dialysis at home.
MATERIALS
Drinking glass Small bowl

Stirring rod 10% NaCl solution
String/rubber bands Starch solution
Betadine Distilled water
Cellophane Scissors
Scoring rubric
For Procedures:
For Rationalization:
For hypothesis/Generalization/Conclusions:

o 3 – all concepts were mentioned and a clear hypothesis, generalization or conclusion was
derived from the activity
o 2 – one or 2 concepts were not mentioned and the hypothesis, generalization or conclusion is
not clear
o 1 – 3 or more concepts were not mentioned and the hypothesis, generalization or conclusion is
not clear
o 0 – no hypothesis, generalization or conclusion was made
THE SET-UP FOR DIALYSIS

1. Obtain a clean cellophane, cut according to size needed and place this atop a clean and
dry evaporating dish/ small bowl.
2. Pour 10 mL of the starch solution and 10 mL of 10% NaCl solution into the cellophane.
3. Gather the ends of the cellophane and tie with a string or rubber band. Make sure that
there are no holes from which the liquid inside can escape from.
4. Tie the cellophane on a pencil/pen and suspend this on a beaker containing distilled water.
The cellophane should be completely submerged but not touching any part of the
beaker/drinking glass. Adjust accordingly.
5. Have your instructor check the set-up (by sending a picture before proceeding to the next
step).
6. Let the set-up stand for 1 hour.
7. Document your set-up and submit this together with your questions for research. Label
the parts correctly.
8. After the required time, remove the cellophane from the beaker.
9. Punch a hole on one side of the cellophane and transfer its content to a clean and dry
beaker/drinking glass. Label this beaker as “A”
10. Save the water in the beaker/drinking glass used for dialysis and label this as “B”.
QUALITATIVE TEST FOR THE SOLUTION INSIDE THE CELLOPHANE
1. Prepare 2 small containers (perfume lid/medicine measuring cup/sauce dish) and place 1
mL of the solution from beaker A into each container.
2. Label the tubes as containers 1 and 2
3. For container 1, should you add 10 drops of conc HNO3 and 1 mL of AgNO3 solution, what
will the result be? Download a picture and attach it to your report.
Observations
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
4. For container 2, add a few drops of betadine.

Observations
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
QUALITATIVE TEST ON THE DIALYSATE
1. Prepare 2 small containers and place 1 mL of the solution from beaker B into each tube.
2. Label the tubes as containers 3 and 4.
3. For container 3, should you add 10 drops of conc HNO3 and 1 mL of AgNO3 solution, what
will happen? Research on the expected result for this procedure.

Observations
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
4. For container 4, add a few drops of betadine.
Observations
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
*Document all your results.
BASED ON THE RESULTS OBTAINED, ANSWER THE FOLLOWING QUESTIONS

1. Were there differences in the results obtained?
___________________________________________________________________
2. Which beaker/drinking glass gave a positive result for the presence of NaCl?
___________________________________________________________________
3. Which beaker/drinking glass gave a positive result for the presence of starch?
___________________________________________________________________
4. What could be the reason for the difference?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
5. Which of the substances placed inside the cellophane is considered a crystalloid?
___________________________________________________________________
6. Which of the substances placed inside the cellophane is considered a colloid?
___________________________________________________________________
7. What did you learn from this activity? Write down your answer on the space provided for
conclusions.
CONCLUSIONS
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
- Solutions in beakers/drinking glass A and B can be poured safely down the drain
- The cellophane is considered as a solid waste and placed in the proper trash can
Scoring Rubric

1. Differentiate between colloids and crystalloids
2. What is a dialysate?
3. Is dialysis the same as osmosis? Explain your answer.
4. What is the physiological importance of dialysis?
5. How does an artificial kidney work?

WEEK NO. 3
SUBMODULE NO. 3
PH AND THE
CHEMISTRY OF
RESPIRATION

LESSON: Unit 3: pH and the Chemistry of Respiration
TOPICS:
1. Definition of terms
1.1. Acids and Bases
1.2. Hydrogen and hydroxyl ions
1.3. Henderson-Hasselbalch equation
1.4.Buffer Substances
2. Chemistry of Respiration
2.1. Respiration and types of respiration
3. Physical theory of respiration
3.1. Chemical Control of respiration
3.2. Oxygen transport
3.3. Carbon dioxide transport
4. The chemical theory of respiration
4.1. Gaseous exchange in the tissues
4.2. Gaseous exchange in the lungs
5. Acid-Base balance
5.1. Buffer system of the blood
5.2. Carbon dioxide elimination through the lungs
5.3. Renal excretion of acids and bases
5.4. Abnormalities of acid-base balance
LECTURE: 6 hours
LABORATORY: 3 hours
LEARNING OBJECTIVES
At the end of the learning session, the student should be able to:
1. Define terms related to pH, and acid base balance.
2. Use correctly the Henderson-Hasselbalch equation in computing for the pH of a
solution.
3. Explain accurately the chemistry of respiration in relation to their course
4. Correctly explain the different buffer systems in the blood.
5. Clearly appreciate how the body compensates for acid base imbalances

LESSON PROPER
INTRODUCTION:
Many people today are interested in exercise as a way of improving their health and
physical abilities. But there is also concern that too much exercise, or exercise that is not
appropriate for certain individuals, may actually do more harm than good. Exercise has many
short-term (acute) and long-term effects that the body must be capable of handling for the
exercise to be beneficial.
When we exercise, our heart rate, systolic blood pressure, and cardiac output (the amount
of blood pumped per heart beat) all increase. Blood flow to the heart, the muscles, and the skin
increase. The body's metabolism becomes more active, producing CO2 and H+ in the muscles.
We breathe faster and deeper to supply the oxygen required by this increased metabolism.
Eventually, with strenuous exercise, our body's metabolism exceeds the oxygen supply and
begins to use alternate biochemical processes that do not require oxygen.
These processes generate lactic acid, which enters the blood stream. As we develop a
long-term habit of exercise, our cardiac output and lung capacity increase, even when we are at
rest, so that we can exercise longer and harder than before. Over time, the amount of muscle in
the body increases, and fat is burned as its energy is needed to help fuel the body's increased
metabolism. (Casiday and Frey, 2008)
Definition of Terms
Assignment: Define the following terms:
1. Ionization –
2. Electrolyte –
3. Strong electrolyte –
4. Weak electrolyte –
5. Weak acid or base –
6. Strong acid or base –
7. Amphiprotic or amphoteric –
8. Buffers –
9. Alkalosis –
10. Acidosis –
11. Ionic equations –
12. Net ionic equation –
13. Spectator ion –
Properties of Acids and Bases
Acids Bases
Turns blue litmus paper to red Turns red litmus paper to blue
Has a sour taste Has a bitter, biting taste
Neutralizes base Neutralizes acids

Reacts with active metals to produce Has s slippery, soapy feeling ( due to
hydrogen gas dissolving a thin layer of skin)
Theories
Arrhenius Concept of Acids and Bases
- Proposed by Svante August Arrhenius (1959 – 1927), a Swedish physicist and chemist
- Defined an acid as a substance that yields hydrogen ions (H+ or H3O+) when dissolved
in water and a base as a substance that yields hydroxide ions (OH-) when dissolved in
water
- Arrhenius suggested that acids are compounds that contain hydrogen and can dissolve
in water to release hydrogen ions into solution
- Arrhenius defined bases as substances that dissolve in water to release hydroxide ions
(OH-) into solution
Bronsted-Lowry Definition of Acids and Bases

- Proposed in 1923 by Johannes Bronsted and Thomas Lowry, working independently
- More general that Arrhenius theory since it is not limited to water solutions
- Defined an acid as any substance that donate a proton (H+) and a base is any
substance that accepts proton (H+)
- any substance that can donate a hydrogen ion is an acid (under the Brønsted definition,
acids are often referred to as proton donors because an H+ ion, hydrogen minus its
electron, is simply a proton).
- The Brønsted base is defined as any substance that can accept a hydrogen ion
- the Brønsted-Lowry definition also explains why substances that do not contain OH- can
act like bases.

- Does not contradict the Arrhenius definition of acid and base; an Arrhenius acid is a
Bronsted-Lowry acid that has dissociated in water by transferring its proton to the
solvent
H+A- + H2O A- + H3O+
- An Arrhenius base (OH-) is also a Bronsted-Lowry base because it accepts a proton in

neutralization
- The difference is that it is no longer the only base that can exist in water. Any species,
molecular or ionic, as long as it can accept a proton, whether in solution or in gas phase
reaction, is considered a base
- When acid donates a proton to water, it becomes a potential proton acceptor and
therefore a base. When water accepts a proton, it acts as a base and becomes a
potential acid
- Conjugate acid-base pair – an acid and a base that are related by a transfer of proton
Common-Acid Base Pair

Strongest Acid Base Weakest
-
HClO4 ClO4
H2SO4 HSO4-
HCl Cl-
HNO3 NO3-
H3O+ H2O
HSO4- SO4-2

H3PO4 H2PO4-
HF F-
CH3COOH CH3COO-
H2CO3 HCO3-
H2S HS-
NH4+ NH3
HCO3- CO3-2
H2O OH-
Weakest OH- O-2 Strongest
LEWIS DEFINITION OF ACIDS AND BASES

- Proposed by G.N. Lewis in 1923
- Most general definition and includes all substances that are acids and bases according
to Arrhenius and Bronsted –Lowry concept
- Defines an acid as any species that can accept a pair of electrons, and a base is any
species that can donate a pair of electrons
- In 1923 G. N. Lewis suggested another way of looking at the reaction between H+ and
OH- ions. In the Bronsted model, the OH- ion is the active species in this reaction it
accepts an H+ ion to form a covalent bond. In the Lewis model, the H+ ion is the active
species it accepts a pair of electrons from the OH- ion to form a covalent bond.
ACTIVITY: identify whether the following describes: (A) Arrhenius theory; (BL) Bronsted-
Lowry and (L) Lewis theory.
_____1. Increases the H3O+ concentration in aqueous solution

_____2. Accepts electron pair
_____3. Accepts protons
_____4. Increases OH- concentration in aqueous solutions
_____5. Donates protons
_____6. Donates electron pairs
Henderson-Hasselbalch equation
- The Henderson-Hasselbalch equation is useful for estimating the pH of a buffer solution

and finding the equilibrium pH in an acid-base reaction.
- The equation can be used to determine the amount of acid and conjugate base needed
to make a buffer solution of a certain pH.
- The Henderson–Hasselbalch equation mathematically connects the measurable pH of a
solution with the pKa (which is equal to -log Ka) of the acid. The equation is also useful
for estimating the pH of a buffer solution and finding the equilibrium pH in an acid-base
reaction. The equation can be derived from the formula of pKa for a weak acid or buffer.

A modified version of the Henderson-Hasselbalch equation can be used to relate the pH
of blood to constituents of the bicarbonate buffering system:
pH = pKa H2CO3 + log [HCO3-]
[H2CO3]
Where:
pKaH2CO3 = acid dissociation constant of carbonic acid which is equal to 6.1
[HCO3-] = the concentration of bicarbonate in the blood
[H2CO3] = the concentration of carbonic acid in the blood
The chemistry of respiration

- Respiration – exchange of gases between the outside air and the body
- Tissues must be adequately supplied with oxygen. Why?
- At rest: 250 mL of oxygen is used and 200 mL of carbon dioxide is eliminated
Composition of Inspired and Expired Air

Oxygen % Carbon Dioxide % Nitrogen %
Inspired 20.95 0.04 79
Expired 16.20 4.38 79
Difference 4.92 4.34 00
The body takes up 5% oxygen and give up 4% CO2 because some oxygen are utilized
for biological oxidation and formation of water
Nitrogen remains the same because nitrogen is supplied by nitrogenous organic
compounds ( proteins, amino acids etc.)
Types of respiration
- External Respiration – the exchange occurring between the outside air and the venous
blood through the lungs
- Internal respiration – the exchange occurring between the blood and the tissues

Physical theory of respiration
Diffusion
- the law that governs the exchange of gases between the outside air, the blood and the
different tissues of the body
- gas flows from a higher to a lower tension
Tension
- the pressure exerted by gas in solution
- Denoted by the symbol p
- e.g. pCO2 – pressure of the dry gas of CO2 in mmHg with which dissolved carbonic acid
in the blood is in equilibrium
pO2 – pressure of the gas with which dissolved oxygen in the blood is in equilibrium
- The tension exerted by an individual gas in a mixture of gases is obtained by multiplying
the total pressure (760 mmHg at sea level) by the percentage of gas in question
To compute for the oxygen tension (pO2):

pO2 = 260 mmHg x 20.96 = 159 mmHg
Try computing for pCO2 = ?
Oxygen tension in different parts of the body in mmHg
Inspired air = 159 – 160

Alveolar air = 108
Venous blood = 40 – 50
Arterial blood = 100
Tissues = 20 – 50
Note that the alveolar pressure is 108 mmHg and the venous blood pressure is 40
mmHg. The difference in pressure serves to drive the oxygen from the lungs to the
blood.
Also note that the partial pressure of oxygen in arterial blood is 100 mmHg. This
blood is carried into the tissues with a partial pressure of only 20 – 50 mmHg
Conditions that affect the diffusion of oxygen from alveolar to venous blood
- Size of the epithelial wall – the combined thickness of the capillary wall and the
respiratory epithelium is not more than 0.004 mm
- The speed of the flow of blood – brings all RBC’s in contact with alveolar air
- The affinity of oxygen to hemoglobin
pCO2 in the various parts of the body in mmHg

- Tissues = 50 – 70
- Arterial blood = 40
- Venous blood = 46
- Alveolar air = 36

- Expired air = 20
- *Atmospheric air = 0.3
• The difference of 10mmHg between venous blood and alveolar air is

enough to drive the CO2 from the blood to the lungs due to the rapidity of diffusion
of CO2 which is 30 times that of O2
THE CHEMICAL CONTROL OF RESPIRATION
- Exerted by the respiratory centers in the medulla and upon the chemical receptors
located at the bifurcation of the common carotid arteries and at the arch of the aorta
Rate and depth of respiration
Carbon dioxide
- Main factor that regulate the rate and depth of respiration
- Increase CO2 in blood = increase rate and depth of respiration leading to increased
pulmonary ventilation
- Leads to immediate elimination of CO2
- Decrease CO2 in blood = slow and shallow respiration leading to decreased pulmonary
ventilation
- Leads to decreased elimination of CO2
EFFECT OF pH
CO2 is low; pH is high = normal rate and depth of respiration
CO2 is low: pH is low = decreased ventilatory rate
- Low oxygen pressure in arterial blood = depression of the respiratory

center, but stimulation of the carotid and aortic chemoreceptors
- The stimulation of the carotid and aortic chemoreceptors causes the
stimulation of respiration and increased ventilation
- Important in the physiological adjustment to low atmospheric
oxygen tension during an ascent to high altitudes
Oxygen transport
- 0.2 – 0.3% - amount taken by the blood plasma when exposed to alveoli air
- Whole blood will take up around 70 – 80 times the amount that the plasma will take
- The oxygen carrying capacity of blood is a function of hemoglobin concentration
- Hemoglobin forms a reversibly stable complex in which the iron remains in the ferrous
state
Hb + O2 HbO2
- 96% - oxygen saturation of the RBC’s leaving the lungs

- 64% - oxygen saturation of venous hemoglobin

- 32% - oxygen that is delivered in the tissues
Since there are around 15 g of hemoglobin per 100 mL of blood and each
gram can combine with 1.34 mL of oxygen (Hufner factor), then 6.4 mL of
O2 has been supplied to the tissues for every 100 mL of blood passing
through the capillaries
• 1.34 X 15 X 0.32 = 6.4 mL of O2
DISSOCIATION OF OXYHEMOGLOBIN
Factors Affecting the Dissociation of Oxyhemoglobin
1. Low oxygen pressure - low pO2 will increase dissociation of oxyhemoglobin to hemoglobin
and oxygen
2. High CO2 pressure – increase pCO2 will decrease the affinity of hemoglobin for oxygen
(Bohr’s effect)
- The effect of CO2 reflects the acidity of carbonic acid solutions
- Increased pCO2 decreases the amount of oxyhemoglobin within the RBC’s
- Increased pCO2 is actually the effect of carbonic acid formation with consequent
lowering of pH
- The equilibrium of the hgb-O2 system is altered causing the dissociation curve to shift to
the right (acid side of isoelectric point of hgb side of dissociation curve)
3. low pH – acids other than

carbonic acid (e.g. lactic acid)
increases the dissociation of
oxyhemoglobin
- More marked when there is a
relatively low pO2
4. Rise of temperature – increase

in temperature will increase the
dissociation of oxyHgb
- In warm-blooded animals,
Hgb gives up O2 more
readily when passing from
high to low tension than in
cold-blooded animals
- Patients with high fever
cause the Hgb to release O2
to tissues
https://www.google.com/search?q=effect+of+co2+bohr+effect&tbm
5. Presence of electrolytes – at low pO2, Hgb gives up O2 more readily in the presence of
electrolytes that it does in pure solution
Carbon dioxide transport

- 50 volumes % - CO2 content of arterial blood
- 55 – 60 volumes % - CO2 content of venous blood

• Each 100 mL of blood transports 5 – 10 mL of CO2 from tissues to lungs
- Three forms of CO2 that is carried in the blood
o 1. As dissolved H2CO3 (5%)

o 2. As carbamino bound CO2 (20%)
o 3. As bicarbonate combined with cations, Na and K (75%)
As H2CO3
- This form is important because any change in its concentration will caused marked
alteration in the blood pH
- Occurs in very small amounts
As carbamino compound
- This is formed with the proteins especially hemoglobin
- The free amino group of the protein reacts with CO2
-
R-NH2 + CO2 R-NH – COOH
- This is an important factor in the physiological efficiency of the respiratory cycle due to
its high rate of reversibility
- The difference of the CO2 between arterial and venous blood is due to the difference in
the carbamino bound form of CO2
- Arterial blood – more oxyHgb favors the release of CO2
- Venous blood – less acid reduced Hgb favors the Hgb – CO2 combination
o Venous blood carries more CO2 in the form of carbamino compound than arterial
blood
As HCO3-
- As CO2 enters the RBC, the CO2 is hydrated to carbonic acid using carbonic anhydrase
(CA). This enzyme also causes its reverse dehydration
CO2 + H2O C.A. H2CO3
- At normal pH, most of the H2CO3 is present in the form of HCO3-
H2CO3 H+ + HCO3-
Carbonic anhydrase
Catalyzes the synthesis of H2CO3 from CO2 and H2O and the degradation of H2CO3
into H+ and HCO3-
Henrique (1928) discovered in the enzyme in RBC’s
It is also found in the muscles, parietal cells of the stomach participating in HCl
production and in the kidney tubules catalyzing hydrogen excretion
Effect of CO2 on blood
- H2CO3 is carried in the blood in the form of bicarbonate

H2CO3 H+ + HCO3-

- At pH 7.4, a ratio of 20:1 must exist between bicarbonate and H2CO3
- As long as this ratio is maintained, the pH of the blood will be normal

- Any alteration n the ratio will disturb the acid-base balance of the blood
- Venous blood carries more CO2 than arterial blood

- The pH of venous blood is more acid by only 0.01 to 0.03 units (pH 7.4 vs pH 7.43)
Blood buffers
1. Plasma proteins – they release sufficient cations to account for the carriage of about
10% of the total CO2
2. Phosphates – within red blood cells – responsible for about 25% of the total CO2
carried
3. Hgb and oxyhemoglobin – accounts for 60% of the CO2 carrying capacity of blood
- Hemoglobin is able to react with protons or dissociate to yield protons
THE CHEMICAL THEORY OF RESPIRATION
Points to remember
1. The wall of the RBC is permeable to water, CO2, H2CO3, Cl- and H+ but not the Hgb
and plasma proteins and only slightly to Na+ and K+
2. Most of the Na ions are in the plasma, while those of potassium are in the cells
3. In the RBC, most of the proteins (Hgb) are combined with K, the amount varying in the
different stages of the cycle
4. the carbonic anhydrase in the RBC hastens the transformation of CO2 and water into
carbonic acid and vice versa
Gaseous exchange in the tissues
A. Carbon dioxide exchange

- CO2 is continually produced as a result of metabolic processes
- 5% is taken up by the blood
- In the RBC, 20% of the CO2 combines with Hgb forming carbamino compounds
- 75% is synthesized into H2CO3
- Increased CO2 = increased H2CO3
- H2CO3 will then ionize into hydrogen and carbonic acid ion
H2CO3 H+ + HCO3-
- HCO3- accumulates in the RBC, the concentration of which will become higher than the
plasma
- What will happen?
o First chloride shift (Hamburger’s phenomenon)
▪ To maintain electrolytic equilibrium, the Cl- ions of the plasma will diffuse
into the red blood cells
B. Oxygen exchange
- oxyHgb (in the form of K oxyhemoglobinate) gives up O2 to tissues to become reduced
Hgb
- Reduced Hgb is a weaker acid and will require less K

- Some K will be available for combination
https://www.google.com/search?q=chloride+shift&sxsrf
CHLORIDE SHIFT
K + HCO3 KHCO3
K + Cl KCl
Na + HCO3- NaHCO3 (happens in the plasma when NaCl released the
Cl- to diffuse into the RBC)
Two opposing phenomena in the oxygen exchange:
1. H2CO3 tends to lower the pH within the RBC

2. The transformation of oxyHgb to reduced Hgb involves a change of pK from 6.2 to 7.7
which tend to rise the pH in the cell
The net result of these processes is to maintain the pH essentially unchanged

Isohydric effect – the shift that takes place at a constant pH
- The combined result of the isohydric effect and the chloride shifts is to increase the
effective osmotic pressure within the cells
- Water is then redistributed between the cells and the plasma
- Hematocrit (the relative volume occupied by RBC’s) is higher in arterial blood compared
to venous blood
o 45 – 49 volume %
GASEOUS EXCHANGE IN THE LUNGS
A. Carbon Dioxide Exchange

- CO2 found in plasma and RBC diffuses into the alveoli to the outside air
- Carbamino compounds dissociates to release CO2

- Carbonic anhydrase splits H2CO3 to CO2 and H2O
H2CO carbonic anhydrase H2O + CO2
B. Oxygen Exchange
- Oxygen combines with reduced Hgb to produce oxyHgb
- oxyHgb is more acidic thus it takes up K to buffer it
- K is liberated from Cl and HCO3- = this hastens the release of CO2
- HCO3- is rapidly lost in RBC’s, HCO3- from the plasma diffuses into the RBC’s
o What will happen?
▪ Second Chloride shift – Cl- goes out from the RBC’s to maintain
electrolytic balance
HEMOGLOBIN
HgbO2 + H+ HHgb + O2
Hemoglobin H
- Abnormal Hgb consisting of 4 B-chains
- Has oxygen affinity 10x normal Hgb
- Does not exhibit Bohr’s effect
- The conformation of the entire Hgb molecule contribute to the affinity of Hgb for
oxygen
Fetal Hemoglobin (HgBF)
- As compared to HgBA (adult), in addition to 2 alpha chains, it has 2 gamma chains
instead of the 2 beta chains
- The ϒ-chain contains isoleucine and a single sulfhydryl group
- This structure shows high affinity for oxygen
- Compared to maternal blood which is 33% saturated, fetal blood has 58% saturation
at 30 mm of O2, 37C and pH 6.8
- HgBF normally disappears after 4 – 6 months except in anemic conditions
What happens when HgBF is treated with an alkali?
Myoglobin
- A hemoprotein capable of reversibly binding oxygen found in muscles
- Exhibits Bohr’s effect
- Venous pO2 of 40 mmHg Hgb = saturated 64% Mgb = saturated 94%
- pO2 of 10 mmHg Hgb = saturated 10% Mgb = 80% saturated
- MgB can accept O2 from HgB and store it in the muscle and release it to cytochrome
oxidase when O2 supply becomes inadequate
- O2 remain fixed to Mgb when the muscle is at rest
- When pO2 falls, O2 dissociates and becomes available for oxidation
- Mgb is abundant in the cardiac muscle, muscle of diving mammals and flight muscle of
birds
Acid – base balance
- Depends on the 1:20 ratio of carbonic acid to bicarbonate

- Acids produced during metabolic activities: carbonic acid, sulfuric, phosphoric and
organic acids

- H2CO3 – principal acid produced during tissue oxidation
- Phosphoric and sulfuric acid – comes from sulfur and phosphorus containing proteins
- Fruits and vegetables (rich in + radicals like Na, K, Ca), can release potentially basic
substances
Factors that maintain normal pH in the body

1. Buffer systems in the blood
2. CO2 elimination through the lungs
3. Renal excretion of acids and bases
4. Renal formation of ammonia – base conservation
Buffer system of the blood
A. Buffer action of hemoglobin

- The buffering effect is due to the imidazole group of histidine
- Exerts is buffering effect on H2CO3
H2CO3 H+ + HCO3-
- H+ is buffered by Hgb and HCO3- diffuses out in the plasma in exchange for chloride
B. Buffer action of bicarbonate

- Bicarbonate is the major buffer of the extracellular fluid
- Exerts buffering effects on fixed acids
- There is more bicarbonate found in the extracellular fluid than other buffer component
- There is limitless supply of CO2
- There are physiological mechanisms which maintain the extracellular pH function by
controlling the bicarbonate or CO2 concentration
- The HCO3-/H2CO3 buffer operates in conjunction with Hgb
- As long as adequate amount of bicarbonate is present, it can dispose of the fixed acids
efficiently
o Alkali reserve – the amount of bicarbonate that serves as a measure of the alkali
available for neutralization of fixed acids
- True plasma
o as pCO2 is increased there is an appreciable increase HCO3- so that the pH
does not fall so rapidly
o As the pCO2 decreases, the HCO3- also decreases thus preventing the
expected rise in pH
Carbon dioxide elimination through the lungs

Factors:
1. The sensitivity of the respiratory center to slight changes of CO2 pressure and the H+
concentration
- If CO2 in increased by 1-5 mmHg it will increase pulmonary ventilation eliminating the
excess CO2; same for H+
- Low pCO2 and H+ leads to hypoventilation, CO2 is retained until it comes back to
normal.
2. The diffusibility of CO2 from the blood thru the pulmonary epithelium into the alveoli air
Renal excretion of acids and bases

- If the fixed acids that have been temporarily neutralized by the HCO3-, are not
eliminated, it will be taken cared of by the kidneys.
- Urinary pH is about 6.0 and the pH of blood is 7.45 – the difference in pH represents the
amount of acid the kidneys remove from the blood
- Urinary pH may vary from 4.5 – 8.2 – our kidneys can excrete excess acids and bases
making it the most important regulator of acid base balance
- Uses HPO4/H2PO4
Renal formation of ammonia
- Urinary ammonia is formed in the distal portion of the uriniferous tubules: the site of
acidification of urine
- Ammonia is derived from the amide grp of glutamine (60%) and from amino acids (40%)
- Factors that stimulate the formation of ammonia
o Increased hydrogen ion concentration
o Diminished bicarbonate of the blood and glomerular filtrate
NH3 + H NH4+
- This prevents the accumulation of H+ and allows continued exchange of H+ with Na+
- The amount of Na+ reabsorbed in the distal tubule reflects the amount of both H+ and
NH4+ ions in the urine
- Thus the fixed alkali (Na+) of the blood is conserved.
- Renal secretion of ammonia is more significant than acidification
- Failure of formation of NH3 may be the cause of acidosis and dehydration in the lower
nephron nephrosis
- Alterations of the carbonic acid factor calls for compensation by the kidneys
- Alterations of the bicarbonate factor calls for compensation by the lungs
Abnormalities of acid-base balance
Metabolic acidosis
- Produced whenever the available base/alkali reserve (Na or K) is decreased although
the total base may remain unchanged

- Conditions that may cause metabolic acidosis: diabetic acidosis, fasting, starvation,
severe diarrhea, kidney failure or uremia (the retention of anions PO4 and SO4 also
decreases the available base)
- Compensation: increased respiration (acidotic breathing) to eliminate more CO2
o As the hyperpnea resulting from the acidosis becomes less, as acidosis
improves, so the stimulus for correction disappears before complete
compensation = the pH remains low, the CO2 combining power is less
Respiratory acidosis (Hypoventilation)

- Occurs whenever there is interference with the exchange of gases within the lungs so
that CO2 is not adequately blown off
- If CO2 is retained, the carbonic acid concentration increases in the blood serum
- The ratio of H2CO3: HCO3- favors the carbonic acid = elevated H concentration and
acidosis
- Conditions that may lead to respiratory acidosis: narcosis from drugs, CNS
depression, emphysema, bronchiectatis, poliomyelitis (during paralysis of respiration)
- Compensation: kidneys will attempt to readjust by excreting more ammonia and
chloride, this conserves Na+
o Na+ becomes available for combination with HCO3- to increase the alkali reserve
o Compensation is usually incomplete
o The ratio remains in favor of the carbonic acid
o Result: low pH with high CO2 combining power
- Management:
o Requires more efficient removal of CO2
o Cautious administration of HCO3- and increased excretion of ficed anions
Respiratory alkalosis (hyperventilation)

- Produced in nay condition causing hyperventilation, when this is not the result of
interference with the gaseous exchange in the lungs
- Increased ventilation blows off large amounts of CO2
- Plasma carbonic acid concentration is decreased
- The ration between HCO3/H2CO3 is decreased , H+ conc is lowered, pH remains high,
- CO2 combining power is normal or lower
- Conditions that may lead to respiratory alkalosis: among nervous patients who are
breathing deeply and rapidly due to some frightening symptom or situation, high fevers,
CNS lesions and anoxia of the cardiac type due to high altitudes
- Compensation:
o Kidneys excrete more bicarbonate
o This improves the ratio of HCO3: H2CO3 but not completely (remains in favor of
the bicarbonate
FORMATIVE ASSESSMENT:
_________________________1. How much CO2 is eliminated by the body at rest?
_________________________2. Law that governs the movement of gases in different tissues
of the body.
_________________________3. The total pressure of mixed gases at sea level.
_________________________4. What is the combined thickness of the capillary wall and the
respiratory epithelium?
_________________________5. A substance present in blood to which oxygen has a strong
affinity.

_________________________6. What is the pCO2 in alveolar air?
_________________________7. Name the location for the chemical receptors for respiration
_________________________8.
_________________________9. What will happen to the rate and depth of respiration when the
CO2 concentration in the blood is increased?
_________________________10. What happens to the rate and depth of respiration if the pH
is increased and CO2 concentration is low?
_________________________11. What will happen to the respiratory center in the medulla if
there is low pO2 in the arterial blood?
_________________________12. What will happen to the ventilatory rate if the pH is low as
well as the pCO2?
_________________________13. Which gas regulates the rate and depth of respiration?
_________________________14. What is the amount of oxygen supplied by 100 mL of blood
in tissues?
_________________________15. What will happen to the rate of dissociation of
oxyhemoglobin if there is low oxygen pressure?
_________________________16. The increase in pCO2 will decrease the affinity of Hgb for
oxygen is known as _____.
_________________________17. What will happen to the rate of the dissociation of
oxyhemoglobin when there is a decrease in temperature?
_________________________18. How much CO2 is transported by blood from tissues to the
lung?
_________________________19. An increase in this form of CO2 in the blood will increase the
pH.
________________________20. Which part of the hemoglobin reacts with CO2 to form
carbamino compounds.
________________________21. An enzyme that causes the hydrolysis of carbonic acid.
________________________22. What should be the ration between HCO3-/H2CO3 in order to
maintain the pH in the blood.
________________________23. What will happen to the amount of H2CO3 if there is also an
increase concentration of CO2?
________________________24. Another name for the first chloride shift.
________________________25. The first chloride shift happens when this ion accumulates in
the red blood cells.
________________________26. This is a shift that happens at constant pH.
________________________27. What is the O2 sat of fetal blood at 30 mm of O2, 37C and pH
6-8?
________________________28. To which enzyme will the Mgb release the oxygen when
oxygen supply becomes inadequate?
________________________29. Organ that serves as the most important regulator of acid-
base balance.
________________________30. In which part of the nephron is the urinary ammonia formed?
ASSIGNMENTS
- Answer the assignments and activities given in this unit
SUMMATIVE ASSESSMENT:
Scoring Rubrik
4 3 2 1

Restate the Question Restated the question Restated almost all Attempted to Did not restate the
completely parts of the restate the question at all
question question but was
unsuccessful
Answer Considered all parts Considered all Missed part of the Did not answer the
of the question and parts of the question question at all
answered each part question but had
accurately only partial
accuracy
Citation of evidence Properly cited Cited evidence Evidence used No evidence from
from texts adequate evidence loosely related to was wither not the texts was used
from the text that the answer related to the
supported the answer question, or not
correctly cited
Elaboration Made connection with Made a connection Attempted to make Did not make a
(connections and the text and clearly to the text, but was a connection to the connection to the
further explanation) explained in unable to explain text but the text at all; element
relationship to the its relation to the relationship was was not present
question text clearly weak
Organization, Response has clear Writer’s response Response needs Response need
Grammar and beginning, middle and flows, but the transition. stronger transition.
punctuation end. Grammar and beginning, middle Grammar and Grammar and
punctuation make the and end is not punctuation needs punctuation makes
response readable clear. Grammar improvement. the text difficult to
and punctuation understant
slows readability
CASE:
Sherry P. Moore is a 68-year-old woman who is admitted to the hospital emergency
room with very low blood pressure (80/40 mmHg) cause by an acute hemorrhage from a
previously diagnosed ulcer of the stomach. Sherry’s bleeding stomach ulcer has reduced her
effective blood volume severely enough to compromise her ability to perfuse (deliver blood to)
her tissues. She is therefore, a “low perfuser”. She is also known to have chronic obstructive
pulmonary disease (COPD) as a result of 42 years of smoking two packs of cigarettes per day.
Her respiratory rate is rapid and labored, her skin is cold and clammy, and her lips are slightly
blue (cyanotic). She appears anxious and moderately confused.
As emergency measures are taken to stabilize her and elevate her blood pressure,
blood is sent for immediate blood typing and cross-matching, so that blood transfusions can be
started. A battery of laboratory tests are ordered including venous hemoglobin, hematocrit and
lactate levels, and arterial blood pH, partial pressures of oxygen (pO2) and carbon dioxide
(pCO2), bicarbonate and oxygen saturation. Results show that the hemorrhaging and COPD
have resulted in hypoxemia, with decreased oxygen delivery to her tissues, and both a
respiratory and metabolic acidosis.
Questions:
1. What is the immediate cause of the confusion experienced by Sherry in the emergency
room? Explain using metabolic reactions and processes.
2. How does Sherry’s preexisting COPD contributed to her hypoxemia?
3. Hoe does Sherry’s preexisting COPD contributed to her respiratory and metabolic
acidosis?

LABORATORY
INTRODUCTION :
All biological processes are greatly affected by the hydronium ion concentration in the
medium in which they occur. Hydronium ion concentration is commonly expressed in terms of
pH.
The maintenance of an optimum pH in the environment of an animal cell is vital to the

life of an organism. The control of pH essential for enzyme – catalyzed reactions is the function
of buffers.
A buffer is a mixture of weak acid and its conjugate base or a weak base and its
conjugate acid. PH of buffer solutions can be determined by using the Henderson – Hasselbach
equation.
Name: _____________________________ Date: _____________________________

Activity No. 5
pH DETERMINATION
OBJECTIVES

1. identify the different indicators used to determine the pH of a solution accurately
2. use confidently different methods to determine the pH of a given solution
3. appreciate wholly the importance of knowing the pH of solution
MATERIALS: Reference Materials
PROCEDURES AND OBSERVATION
Scoring rubric
For Rationalization
- 3 points is given for every hypothesis, generalizations or conclusions made
o 2 – one or 2 concepts were not mentioned and the hypothesis, generalization or conclusion
is not clear
o 1 – 3 or more concepts were not mentioned and the hypothesis, generalization or conclusion
is not clear
o 0 – no hypothesis was made

DETERMINATION OF pH VALUES USING CHEMICAL INDICATORS
1. What would be the color change of methyl red if placed in acidic

solution_____________? In a basic solution_______________?
2. What would be the color change of phenolphthalein if placed in an acidic
solution____________? In a basic solution______________?
3. What would be the color change of bromothymol blue if added to an acidic
solution_______________? In a basic solution_______________?
4. What would be the expected color change if an acid is reacted with blue litmus paper?
_____________
5. With red litmus paper? _______________

6. What would be the expected color change if a base is reacted with blue litmus paper?
______________
7. With red litmus paper?_________________
FILL-UP THE TABLE BELOW.
TEST METHYL PHENOLPHTHALEIN BROMOTHYMOL ACID LITMUS

SUBSTANCES RED BLUE OR PAPER
BASE
Calamansi
juice
Coke
Vinegar
Antacid tablet
1% Na2CO3
1. What is the principle behind the use of pH paper?

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
2. What is the principle behind the use of pH meters?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
3. What is the principle behind the use of chemical indicators?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
4. What are the limitations of using chemical indicators in determining the pH of a given
solution?

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
5. Name at least 5 household materials that can be used to determine the approximate pH
of substances and indicate their color change.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Scoring Rubric
1. Discuss the Acid-Base Balance Mechanism of the human body.

2. Differentiate the following Acid-Base imbalances in terms of results in Blood Gas
Analysis and possible causes:
a. Metabolic Acidosis
b. Respiratory Acidosis
c. Metabolic Alkalosis
d. Respiratory Alkalosis
3. What is the importance of determining the pH of body fluids of individuals?
4. You have a patient with urinary tract infection or UTI. Will it be useful to determine
the pH of the urine of your patient? Why?

Week no. 4
Submodule no. 4
Nucleic acids and

heredity

LESSON: NUCLEIC ACIDS
TOPICS:
1. Definition, Importance and Functions of Nucleic Acids
2. Structural Components, Nomenclature and Features
2.1. Nucleoproteins
2.2. Nucleotides
2.3. Nucleosides
3. The DNA molecule
3.1. DNA Structure DNA Replication
3.2. The RNA molecule
4. RNA Structure
4.1. Types of RNA
5. The Genetic Code
5.1. Protein Synthesis
5.2. Transcription
5.3. Translation
6. Mutation and other Clinical Significances of Nucleic
TIME/SCHEDULE: Week no. 4
LECTURE: 6 hours
LABORATORY: 3 hours
LEARNING OBJECTIVES
At the end of the leaning session, the student should be able to:
1. Describe accurately the nucleic acids at the molecular level
2. Differentiate completely DNA from RNA
3. Appreciate the intricate structure of the nucleic acids
4. Explain completely the process of transcription
5. Illustrate simply the process of translation
6. Relate the importance of this unit to mutation and the significance of nucleic acid
metabolism
7. Perform laboratory procedures correctly

LESSON PROPER
INTRODUCTION:
In the middle to the 20th century, DNA was identified as the genetic material and its
structure was determined. Using this knowledge, researchers then discovered the mechanisms
by which genetic information is inherited and expressed. During the last quarter of the 20 th
century, our understanding of this critical area of science, known as molecular biology, grew at
an increasingly rapid pace. We now have techniques to probe the human genome that will
completely revolutionize the way medicine is practiced in the 21 st century.
Research in molecular biology has produced a host of techniques, known collectively as
recombinant DNA technology, biotechnology, or genetic engineering, that can be used for the
diagnosis and treatment of diseases. These techniques can detect a number of genetic
disseases that previously could only be diagnosed after the symptoms appeared. Diagnosis of
these diseases can now be made with considerable accuracy even before birth, and carriers of
these diseases can also be identified. (Lieberman, Marks, Smith, 2005)
TYPES OF TRAITS
1. DOMINANT TRAIT
- it is one which is present or evident or manifested in majority of the offspring or children
in every generation
- e.g. black hair is dominant over blond hair
2. RECESSIVE TRAIT
- is one which may be seen only in a minority of offspring’s
- the trait may even disappear in one generation but will re-appear in succeeding
generations
Activity:
Identify the following human traits as to dominant (D) and Recessive (R)
_____1. Cleft chin _____5. Freckles
_____2. Widow’s peak _____6. Blue eyes
_____3. Dimples _____7. Attached earlobes
_____4. Blond hair
TYPES OF EXPRESSION OF HEREDIATRY TRAITS
1. PHENOTYPES
the physical observable aspects of heredity as handed down from parents to offspring’s
e.g. color of eyes, bridge of the nose, dimples
2. GENOTYPE
the non-observable, non-physical aspects of heredity
e.g. IQ, talent, allergy (asthma), diabetes, epilepsy, hemophilia, insanity, color blindness
NUCLEOPROTEINS
- Conjugated proteins
- Proteins + nucleic acids
- Sources
o Tissues with closely packed cells and big nuclei

▪ Thymus, liver, spleen, kidney and pancreas
o Found in bacteria, bacteriophage, chromosome and simple filterable viruses
- Protein component: simple proteins, albumins, histones or protamines
- Constitute a large part of the nuclear material of the cell
- Also found in the cytoplasm, particularly in the ribosome (concerned with protein
synthesis)
PROPERTIES OF NUCLEOPROTEINS
- Acidic
- Soluble in alkalies with which they form salts
- Precipitated from solutions by acetic acid but are re-dissolved by dilute HCl
- Not coagulated by heat
- Exhibit similar precipitation and color reactions with proteins
HYDROLYSIS OF
NUCLEOPROTEINS
https://www.google.com/search?q=hydrolysis+of+nucleoproteins&tbm
IMPORTANCE OF NUCLEOPROTEINS
- They are closely associated with chromosomes
VIRUSES AS NUCLEOPROTEINS
- 1935 – W.M. Stanley isolated the virus that
causes tobacco mosaic disease
- 1986 – Bawden and Pirie – the virus that
causes tobacco mosaic disease is a
ribonucleoprotein
VIRUSES
- Can reproduce inside living cells but are
incapable of doing so independently
- Nucleoproteins have the ability to reproduce
but should be within a host cell
- The RNA alone possesses biological activity,
the protein serve as coat
https://www.google.com/search?q=tobacco+mosaic+virus&t
bm

BACTERIOPHAGES
- Viruses of bacteria
- The protein helps in the penetration
of the bacterial cell
- Only the nucleic acid enters the cell
ANIMAL VIRUSES
- They are classified by containing
either DNA or RNA
https://www.google.com/search?q=bacteriophage&tb
https://www.google.com/search?q=bacteriophage&tb
m=isch&hl=en&chips
TEST YOUR KNOWLEDGE:

- Classify the following diseases as being caused by (A) DNA and (B) RNA viruses
_____1. Chickenpox ____4. Rabies
_____2. Warts ____5. Hepatitis B
_____3. CoViD 19 ____6. AIDS
https://www.google.com/search?q=the+influenza+virus&tbm=isch&ved=
THE INFLUENZA VIRUS

NUCLEIC ACIDS
- Macromolecules
- first discovered in the nuclei of cells
- contain all the information to direct the activities of a cell and its reproduction
IMPORTANCE
- essential in understanding genetics
- to understand the many aspects of diseases like their
o Pathophysiology
o genetic basis of the disease
Functions of Nucleoproteins and Nucleic acids

A. To reproduce their own kind (duplication)
B. To store, express and transmit genetic information
C. The undergo mutation
Components
- Elementary components: 15 - 16% N; 9 – 10% P
Properties of Nucleic acids

- Insoluble in alcohol
- Slightly soluble in cold water
- Readily dissolves in hot water and dilute alkalies forming alkali salts
- Precipitated by HCl and excess acetic acid
Molecular anatomy of nucleic acids

- Hydrolysis yields nitrogen base, sugar and phosphoric acid
I. NITROGENOUS BASE
1. Pyrimidine
2. Purine
PYRIMIDINE
- 6-membered ring structure
- Include:
o A. cytosine – 2 – oxy-4-aminopyridine
o B. uracil – 2,4 – dioxypyrimidine
o C. Thymine – 5 – methyl – 2,4 -
dioxypyrimidine
- Methylation – caused by special enzymes called

DNA methytransferase
- example: 5 – methylcytosine
- Other methylations may be induced by carcinogens
https://www.google.com/search?q=
pyrimidine&tbm=isch&ved=2ahUKE
wjjkvnI4IDrAhVxEqYKHWwNB0EQ
2
PRYIMIDINE

http://www.tud.ttu.ee/im/Tonu.Reintamm/shabarova/1.2.html
OTHER BIOMOLECULES THAT CONTAIN THE PYRIMIDINE STRUCTURE:
- 5-hydroxymethylcytosine
o found in bacteriophage
- 5-hydroxymethyluracil
o found in B. subtilis in place of thymine in DNA
- Vitamin B1 – thiamine
o Important in metabolic processes
- Alloxan – 2,4,5,6 – tetraoxypyrimidine
o Produces experimental diabetes
- 2 –thiouracil
o Used for thyroid treatment
- 5 – fluorouracil
o Used for a variety of cancers (breast and digestive)
PURINE BASE
- Includes pyrimidine and imidazole ring
- Include
o Adenine – 6 – aminopurie
o Guanine – 2-amino-6-oxypurine
http://www.brainkart.com/article/Purine-bases--Structure-and-Properties_27796/
OTHER BIOMOLECULES WITH THE PURINE BASE STRUCTURE
- Hypoxanthine – 6-oxypurine
o metabolic by-product
- Xanthine - 2,6-dioxypurine
o Metabolic by-product
- Uric acid – 2,6, 8 –trioxypurine
o Metabolic by-product

- Caffeine – 1,3,7 – trimethylxanthine
o Plant source: coffee
- Theobromine – 3,7 – dimethylxanthine
o Plant source: tea and cocoa
- 6-mercaptopurine
o Used for the treatment of leukemia
II. SUGARS
- Ribose
- Deoxyribose
III. PHOSPHORIC ACID

- H3PO4
- H2O3POH
SUGAR + BASE = NUCLEOSIDE
CLASSIFICATION OF NUCLEOSIDES
NUCLEOSIDE - Uses β – glycosidic linkage

- For purine nucleosides
o C1 of sugar to N9 of purine
- For pyrimidine nucleosides
o C1 of sugar to N1 of pyrimidine
https://www.mikeblaber.org/oldwine/BCH4053/Lecture18/Lecture18.htm

https://quizlet.com/132523292/nucleotide-metabolism-flash-cards/
NITROGENOUS BASE + SUGAR + PHOSPHATE =

NUCLEOTIDE
NUCLEOTIDE
- Basic structural unit of nucleic acids
- It is the monomer for nucleic acids
- Chemically:
o Phosphoric acid ester of nucleoside
o Strongly acidic
- Includes:
o Adenylic acid
o Guanylic acid
o Thymidic acid
o Cytidylic acid
o Uridylic acid
Formation of bonds:
- For deoxyribose
o Phosphorylation at C3 and C5
o C1 and C4 – involved in furanose
ring
o C2 – no –OH group
- For ribose
o Phosphorylation at C2, C3 and C5
o C1 and C4 – involved in furanose
ring formation

https://www.biologyjunction.com/nucleotide_model_preap.htm
https://www.researchgate.net/figure/Adenine-nucleosides-and-nucleotides-structure-Nucleosides-consist-of-a-
nucleobase_fig1_330823477
Names of Nucleosides and Nucleotides in DNA and RNA
Base Nucleosides RNA

Nucleotides
Adenine (A) Adenosine (A) Adenosine monophosphate (AMP)
Guanine (G) Guanosine (G) Guanosine monophosphate (GMP)
Cytosine (C) Cytidine (C) Cytidine monophosphate (CMP)
Uracil (U) Uridine (U) Uridine monophosphate (UMP)
DNA
Adenine (A) Deoxyadenosine (A) Deoxyadenosine monophosphate (dAMP)
Guanine (G) Deoxyguanosine (G) Deoxyguanosine monophosphate (dGMP)
Cystosine (C) Deoxycytidine (C) Deoxycytidine monophosphate (dCMP)
Thymine (T) Deoxythymidine (T) Deoxythymidine monophosphate (dTMP)
Functions of Nucleoproteins and Nucleic acids
- To reproduce their own kind (duplication)

- To store, express and transmit genetic information
- They undergo mutation
2 Types of Nucleic Acids

1. Deoxyribonucleic acid (DNA)
- Main carrier of genetic information
- The basic information pathway
- The chemical basis of heredity
- Yields D-2’-deoxyribose, phosphoric acid and four nitrogenous bases of which are
adenine (A), guanine (G), cytosine (C) and thymine (T)

2. Ribonucleic acid (RNA)
- Aids in expressing genetic characterisctics
- A copy form a portion of the DNA which becomes a template for synthesizing an
intended protein
- Yield ribose, phosphoric acid and four nitrogenous bases namely adenine (A), guanine
(G), cytosine (C) and uracil (U) upon hydrolysis
Comparison of the DNA and RNA
Differences DNA RNA
A. Composition
1. Sugar component Deoxyribose Ribose
2. Pyrimidine bases Cytosine Cytosine

thymine Uracil
3. Purine Bases Adenine Adenine
Guanine Guanine
4. Phosphate (fr H PO ) present Present
3 4
B. Location Cell nucleus Cytoplasm

Mitochondria Ribosomes
Viruses Nucleolus
Viruses
C. Structure Linear Branched
D. Color tests
1. Fuelgen Test Red Negative

2. Dische test Blue Negative
3. Orcinol test Negative Green

4. Aniline Red Negative
THE DNA
- A polymeric substance made up of the four nucleotides (dAMP, dGMP, dCMP, dTMP)
- Size varies with the complexity of the organisms (E. coli = 8 million nucleotides; human =
500 million nucleotides)
- The chemical basis of heredity
- Genome – the sum total of all hereditary material contained in a cell
o Within the genome are chromosomes
o Within chromosomes are genes – fundamental units of heredity
o Gene – segment of the DNA chain that controls the formation of a molecule of
RNA

Structure of the DNA
Internucleotide linkages
- Link that join nucleotides
together
- It is of the diester type
- Between C3 and C5 of
the sugar molecule (3,5-
phosphodiester bridges)
- Double Helical
Structure
- The early 1950’s, James
Watson and Francis
Crick determined that
DNA had a double-
helical structure with two
nucleotide strands winding about each
other like a spiral staircase
NB
- Held by hydrogen bonds
- Amino is joined to a keto group
o A–T 2 hydrogen bonds
o T–A
o G–C 3 hydrogen bonds
o C–G
- Sugar- phosphate outside, bases inside
THE G-C BASE PAIR
THE A-T BASE PAIR
https://www.chemguide.co.uk/organicprops/aminoacids/dna1.html

THE BASIC DNA STRUCTURE
https://teachmephysiology.com/basics/cell-growth-death/dna-replication/
THE DNA STRUCTURE SHOWING HYDROGEN BONDS
https://www.chemguide.co.uk/organicprops/aminoacids/dna1.html
CHARGAFF’S RULES OF THE DNA
- The base composition of the DNA of an organism is constant in all
- The amount of adenine is always equal to the amount of thymine
- The amount of guanine is always equal to the amount of cytosine
- The is a 1:1 purine - pyrimidine ratio

Replication of DNA molecules
DNA Replication – is the biochemical process by which DNA molecules produce exact
duplicates of themselves
- Each time the cell divides, an exact copy of the DNA of the parent cell is needed for the
new daughter cell
Enzymes involved
1. DNA helicase
- causes the DNA helix to unwind
2. DNA ligase
- Joins Okazaki fragments together
o Okazaki fragments – short segments of the DNA molecule
- Forms the other strand of the DNA (3’ to 5’)
- Nicks – gaps in the daughter strand
3. DNA polymerase
- catalyzes the formation of a new phosphodiester linkage between the nuceotide and the
growing strand
- Can operate only in the 5’ to 3’ direction
- Only one strand can grow continuously in the 5’ to 3’ direction
STEPS IN DNA REPLICATION
Step 1
- The enzyme DNA helicase causes the two strands to unwind, producing two separate
strands
Step 2
- Free nucleotides pair with their complementary base on the template strands by means
of hydrogen bonds
Step 3
- DNA polymerase joins the newly attached nucleotides to create one continuous strand in
the 5’ to 3’ direction

Step 4
- The other strand is formed in short segments (Okazaki fragments) in the 3’ to 5’
direction. The segments are joined together by DNA ligase
CHROMOSOMES
- Histone – DNA complexes

- Structural units that provide the
most stable arrangement for the
long DNA molecule
- An individual DNA molecule
bound to a group of proteins
- 15% by mass DNA and 85% by
mass protein
- Different organisms have
different number of chromosomes
o Humans 46; mosquito 6;
frog 26; dog 78; turkey 82
https://sphweb.bumc.bu.edu/otlt/MPH-Modules/PH/DNA-
Genetics/DNA-Genetics2.html
Activity: using a short bond paper, draw a typical human chromosome and label the
parts.
4 major differences between RNA and DNA

- Ribose is the sugar unit in RNA
- Thymine is replaced by uracil in RNA. Uracil pairs with adenine
- RNA is single stranded, thus it does not contain equal amounts of specific bases
- The RNA is much smaller than the DNA (from 75 nucleotides to a few thousand
nucleotides)
THE BASIC RNA STRUCTURE
5 major types of RNA
1. Heterogeneous nuclear RNA (hnRNA)

2. Messenger RNA (mRNA)
3. Small nuclear RNA (snRNA)
4. Ribosomal RNA (rRNA)
5. Transfer RNA (tRNA)
Heterogeneous Nuclear RNA

- RNA formed directly by DNA transcription.
Post - - transcription processing converts the
heterogeneous nuclear RNA to messenger RNA
Small nuclear RNA

- RNA that facilitates the conversion of
heterogeneous nuclear RNA to messenger RNA
- It contains from 100 to 200 nucleotides
https://microbenotes.com/rna-properties-structure-types-and-
functions/

Messenger RNA (mRNA)
- Carries genetic information from the DNA in the nucleus to the ribosomes for protein
synthesis
- Each gene in a DNA produces separate mRNA when certain protein is required by the
cell
- A template made from DNA
- It carries the code that directs the synthesis of proteins
- The size of the mRNA depends of the number of nucleotides in that particular gene
- mRNA + hnRNA = 5 – 10% of total RNA material
Transfer RNA (tRNA)

- The smallest RNA molecule
- Functions
o Interprets the genetic code
o Brings specific amino acids to the ribosome for protein synthesis
o Link the genetic code to the amino acids in the proteins
- There are different tRNA’s for all the amino acids
- Each tRNA consists of a single chain of about 70-90 nucleotides
- Makes up 10 – 15% of cellular RNA
Cloverleaf model of the tRNA

- Anticodon loop – Hydrogen bonds between one of the complementary nitrogen bases in
the chain produce loops that give an overall cloverleaf structure. At the center loop, there
are three bases known as anticodon that are complementary to three bases on an
mRNA codon
o Composed of 7 unpaired nucleotides
o 3 of which make up an anticodon
- Other loops enable the tRNA to bind to the ribosome and other specific enzymes during
protein synthesis
http://ifasonline.com/NEET-AIIMS-COACHING/Biology/Neet-Biology-topic-36.jsp
Ribosomal RNA (rRNA)
- The most abundant RNA
- Makes up of 60% of the structural material of ribosomes

- Ribosomes
o the site of protein synthesis
o small, complex structures in the cytoplasm composed of rRNA and protein
- RNA that combines with specific proteins to form ribosomes , the physical sites for
protein synthesis
- The rRNA present in ribosomes has no informational function
Synthesis of RNA: Transcription

- The transfer of genetic information from DNA by the formation of mRNA
- In this process, which occurs in the nucleus, the nucleotides on one strand of DNA are
copied or transcribed in their complementary bases in a messenger RNA molecule
- However, the entire DNA cell is not necessarily transcribed, only individual genes or
group of genes are copied
- The process is aided by the enzyme RNA polymerase which is able to recognize the
initiating signal and the terminating sequence for a particular gene that needs to be
copied
- Example
Bases on a DNA template strand - G- T- A- C-
Complementary base sequence in mRNA - C- A-U- G-
- Oncogenes – present in cancerous or malignant cells and code for proteins that control
cell growth
o Cancerous cells lose control of oncogene transcription
o Control is lost when DNA is altered
o Oncogenes are transcribed too often, cell growth is uncontrolled – cancer
develops
- Tumor suppressor genes (p53)
o Genes code for proteins that allow cell growth only if the cells are correctly
functioning
- Apoptosis
o Self-destruction of potential cancer cells
o Due to active tumor-suppressor gene
Translation
- The overall function of the rRNA’s in the cell is to facilitate the lack of synthesizing
protein
- After the genetic information encoded in DNA is transcribed into mRNA, it moves out of
the nucleus to the ribosomes in the cytoplasm
- At the ribosome, translation by the tRNA’s converts the genetic information in the
mRNA’s into a sequence of amino acids in proteins

The Central Dogma (by Francis
Crick in 1957)
- Explains the flow of genetic

information within a cell
- Also known as “one gene-one
protein hypothesis”
- The genetic information
contained in one gene of a DNa
molecule is used to make one
molecule of mRNA by a
process known as transcription
- The genetic information in that
mRNA molecule is then used to make one protein by a process known as translation
Protein Synthesis
I. Transcription
- The process by which a segment of the DNA is cut off or nicked from the molecule by
the enzyme - RNA polymerase
- The detached portion together with its accompanying nitrogenous base will serve as a
template for the formation of a complementary strand finally resulting to the formation of
a mRNA (occurs in the nucleus)
- To reconstruct the DNA molecule: OKAZAKI fragments are put together by ligase
- mRNA leaves the nucleus and goes to the ribosomes, it carries with it the codon
II. Translation
- the process by which tRNA accepts the codon carried by the mRNA, reacts, translates
and decodes the message
- pertains to the production of the 20 amino acids
III. Formation of polypeptide Chain

- amino acids in the cytoplasm are activated by combining with ATP
Amino Acid + ATP Amino acyl tRNA sythetase Amino acyl Adenylate
- the tRNA will transport individual amino acids to the ribosomes based on the
arrangement which follows strictly the dictates of the codon
The Genetic Code

- Genetic information from DNA in encoded in the mRNA as a sequence of nucleotides
- The genetic code is the sequence of three bases (or triplet), called a codon, that
specifies the amino acid order for the synthesis of proteins
- Example
Codons (base sequence) in mRNA UUU – UUU – UCA
Translation
Amino Acid sequence Phe – phe – Ser
- Thus, a protein with 200 AA = 200 codons
o 200 x 3 = 600 nucleotides
• Codon – a sequence of three bases in mRNA that specified a certain amino acid to
be placed in a protein. A few codons signal the start or stop of transcription

• Anticodon – the triplet of bases in the center loop of tRNA that is complementary to
a codon on mRNA
Genetic code for Messenger RNA (Introduction to General, Organic and Biochemistry by Hein
8th edition)
First Second Third Nucleotide and Amino Acid Coded
Nucleotide nucleotide U C A G
U Phe Phe Leu Leu
U C Ser Ser Ser Ser
A Tyr Tyr TC* TC*
G Cys Cys TC* Tryp
U Leu Leu Leu Leu
C C Pro Pro Pro Pro
A His His Gln Gln
G Arg Arg Arg Arg
U Ile Ile Ile Met
A C Thr Thr Thr Thr
A Asn Asn Lys Lys
G Ser Ser Arg Arg
U Val Val Val Val
G C Ala Ala Ala Ala
A Asp Asp Glu Glu
G Gly Gly Gly Gly
TC – Termination codon or nonsense codon

- They do not encode any amino acids
- Act as signals to indicate where the synthesis of a protein molecule is to end
Example:
Codon Message (A.A.)
AAA Phenylalanine
AAT Leucine
TAC Methionine
ATA Tyrosine
GTG Histidine
ACC Tryptophan
*64 three-lettered codons (there is more than 1 codon for most amino acids)
Characteristics of the codon

o Universal
▪ All plants, animals and bacterial cell have the same codon to specify each
amino acid
o Degenerate
▪ More than one triplet can code for a particular amino acid
o Continuous
▪ Non-overlapping
▪ Adjacent codons do not overlap

Three Processes involved in the Biosynthesis of Proteins
1. Initiation
a. The codon AUG or GUG (AUG is more common)signals the start of protein
synthesis
b. Once the correct starting point has been identified, a special initiator tRNA binds
to the ribosome
2. Elongation – after the initiator tRNA attaches to the mRNA codon, the elongation of the
polypeptide chain starts
a. The formation of the peptide chain which assembles one amino acid at a time
b. Translocation – the shift of a ribosome along mRNA from one codon (three
bases) to the next codon during translation
c. The process of elongation continues as the ribosome moves along (translocates)
the mRNA, one codon at a time
3. Termination
a. Protein synthesis stops when a ribosome encounters the codon UGA, UAA or
UAG
PROTEIN SYNTHESIS

Genetic Mutations
- A mutation is a change in the DNA base sequence that alters the structure and function
of the protein in the cell
Kinds of Mutation
A. Beneficial
B. Harmful
a. Lethal
C. Silent
Types of Mutation
A. Substitution – a mutation that replaces one base in a DNA with a different base. The
change in the codon may cause a different amino acid to be inserted at that point in the
polypeptide
Substance Normal sequence Effect of mutation

Possible change in base
DNA ACA-CCC-AGG-TTT ACA-CAC-AGG-TTT
Change in codon
mRNA UGU-GGG-UCC-AAA UGU-GUG-UCC-AAA
Change in amino acid order
Amino acid sequence Cys-Gly-Ser-Lys Cys-Val-Ser-Lys
B. Frame Shift Mutation – a mutation that inserts or deletes a base in a DNA sequence
Substance Normal sequence Frame shift effect of Mutation

A is added A is deleted
DNA -CCC-AGG-TTT- -CCC-AAG-GTT-T -CCC-GGT-TT
Shift in codons
mRNA -GGG-UCC-AAA- -GGG-UUC-CAA-A -GGG-CCA-AA
Incorrect order
Amino -gly-ser-lys- Gly-Phe-Gln -Gly-Pro
acid
sequence
Factors that Cause Mutation

- Viruses
- Chemicals (industrial chemicals, pesticides, food additives, hair dyes and cosmetics)
- Ultraviolet light (overexposure to sum)
- X-rays
Mutagen
- The agents that causes mutation which include chemical agents or various types of
radiation
- Ames test – developed by Bruce Ames, is the standard test for mutagenicity

The following shows the effect of mutation:
DNA Mutagen alteration of DNA Defective protein genetic disease

(germ cells)
Cancer
(somatic cells)
Some Genetic Diseases
A. Down’s syndrome – is the leading cause of mental retardation, occurring in about 1 of
every 800 live births. Mental and physical problem including heart and eye defects are
the result of the formation of three chromosomes, usually chromosome 21, instead of a
pair
B. Sickle-cell anemia – a defective
hemoglobin from a mutation in a
gene on chromosome 11
decreases the oxygen-carrying
capacity of red blood cells that
take on a sickle-shape, causing
anemia and plugged capillaries
from red blood cell aggregation
C. Galactosemia – absence of the
transferase enzyme required for
the metabolism of galactose-1-
phosphate, leading to cataracts
and mental retardation
https://xconomy.com/boston/2018/06/15/bluebirds-upgraded-
gene-therapy-shows-promise-for-sickle-cell-patients/
SICKLE –SHAPED RED BLOOD CELLS
D. Cystic fibrosis – the most common inherited disease, thick mucus secretions make
breathing difficult and block pancreatic functions
E. Familial hypercholesterolenemia – a mutation of a gene on chromosome 19
characterized by high cholesterol levels leading to early coronary heart disease in 30-40
year-old persons
F. Muscular Dystrophy (Duchenne) – one of 10 forms of MD caused by a mutation in the
X chromosome occurring in about 1 of 10,000 males due to the low or abnormal
production of dystrophin by the x gene, a muscle-destroying disease beginning at about
age 5 with death by age 20
G. Hemophilia – one or more defective blood clotting factors leading to poor coagulation,
excessive bleeding and internal hemorrhages
H. Tay-Sach’s disease – a defective hexosaminidase A, causing an accumulation of
gangliosides resulting in mental retardation, loss of motor control and early death
I. Huntington’s disease – a genetic disease appearing in middle age affecting the
nervous system that leads to total physical impairment, result of a mutation in a gene on
chromosome 4, which can now be mapped to test people in families with HD

4 Means of Manipulating Genes
A. Genetic Engineering
B. Recombinant DNA technology
C. Genetic manipulation (GM)
D. Gene splicing
• Over the past 25 years, geneticists have been cutting, splicing, and rejoining DNA
from different genes to form a new synthetic DNA called recombinant DNA
Genetic Engineering
- Uses the techniques of molecular cloning and transformation to alter the structure and
characteristics of genes directly
- Most of these experimentation has been done with E. coli, which contains a single
chromosome and several small cyclic DNA particles called plasmids
Some Successful Applications of Genetic Engineering

- Improving crop technology
- Manufacture of synthetic human insulin thru the use of modified bacteria (E. coli
plasmids)
- Manufacture of erythropoietin in hamster’s ovary cells
- Production of new types of experimental mice such as the oncomouse (cancer mouse)
for research
- Created the human growth hormone as a replacement for a drug that was previously
extracted from human cadavers
- In 1987, the FDA approved the first genetically engineered vaccine for humans for
hepatitis B
- The protein interferon, which helps fight viral infections and possibly cancer, has been
produced by recombinant DNA
- Creation of genetically modified organisms (GMO’s)such as foods and vegetables that
resist pest and bacterial infection and have longer freshness than otherwise
- Human genetic engineering can be used to treat genetic diseases
- Human genetic engineering is already being used on a small scale to allow infertile
women with genetic defects in their mitochondria to have children, healthy human eggs
from a second mother are used
- Genetic engineering has the potential to change human beings, appearance,
adaptability, intelligence, character and behavior
How is this accomplished?

A. Isolation of the genes of interest
B. Insertion of the genes into a transfer vector, such as a plasmid, viral vector or
liposomes
C. Transfer of the vector to the organism to be modified
D. Transformation of the cells of the organism
E. Selection of the GMO from those that have not been successfully modified

Nb. As a control measure, the E. coli strain used for research is made deficient in one amino
acid, so that it is dependent on artificial conditions for survival
______________________________1. What is the chemical basis of heredity?
______________________________2. What is the positive color result for Dische test?
______________________________3. It is the sum total of all hereditary material found in the
cell
______________________________4. Chargaff’s rule state that the amount of adenine is
always equal to ____.
______________________________5. In what direction does the enzyme DNA polymerase
operate?
______________________________6. What enzyme cause the DNA helix to unwind?
______________________________7. Nucleoproteins react with what group of substances to
form salts.
______________________________8. What do you call the viruses of bacteria?
______________________________9. What do you call the short segments of DNA molecule
seen during DNA replication?
_____________________________10. What do you call the fundamental units of heredity?
_____________________________11. Which nitrogenous base in present in RNA that is
absent in DNA?
_____________________________12. How many hydrogen bonds are there between guanine
and cytosine?
_____________________________13. It the process of making mRNA from one gene of DNA
_____________________________14. Which enzyme uses a portion of the DNA to be used as
a template during protein synthesis?
_____________________________15. This happens when the DNA base sequence is
changed which alters the structure and function of the protein in a cell
_____________________________16. How many nucleotides make up the anticodon?
_____________________________17. The codon is found in which type of RNA?
_____________________________18. The anticodon is found in which type of RNA?
_____________________________19. Which of mutation results from the insertion or deletion
of a base in the DNA sequence?
_____________________________20. Which of RNA is formed directly by DNA transcription?
Scoring Rubrik for short response assignment and case analysis

4 3 2 1
unsuccessful
accuracy
correctly cited

slows readability
ASSIGNMENT:
1. What is the importance of DNA methylation?

2. What would be the DNA sequence of oxytocin, a hormone that promotes childbirth labor
if its amino acid sequence is as follows: cys-tyr-ile-gln-asn-cys-pro-leu-gly-NH2
3. What property of the A-T rich region of a DNA double helix makes it suitable to serve as
a recognition site for the start point of transcription?
4. Answer all activities found in this unit.
Case 1:
Mary Cruz is a 26 y/o IV drug abuser who admitted sharing unsterile needle with another
addict for several years. Five months before presenting to the hospital emergency department
with soaking night sweats, she experienced a 3-week course of a flu-like syndrome with fever,
malaise, and muscle aches. Four months ago, she noted generalized lymph node enlargement
associated with chills, anorexia and diarrhea, which led to a 22-lb weight loss. Test were
positive of HIV.
Mary Cruz was then treated with a mixture of drugs including zidovudine (ZDV) formerly
called AZT and didanosine (ddl). However, Ms. Cruz was having difficulty complying with her
multidrug regimen; she often forgets to take her pills. When she returns for a check-up, she
asks whether such a large number pills are really necessary for the treatment of AIDS.
Questions:
1. How does the HIV virus work? Answer using what you have learned from this chapter.
2. How does ZDV and ddl work to slow the course of the disease? Your answer must be
relation to the query of Ms. Cruz.
Case 2:
Anne Schukate is a 4 y/o girl of Mediterranean ancestry whose height and body weight
are below the 20th percentile for girls of her age. She is listless, tires easily and complains of
loss of appetite and shortness of breath on exertion. A dull pain has been present in her right
upper quadrant for the last three months. Her complexion is slate-gray and she appears pale.
Initial laboratory studies indicate a severe anemia with a hemoglobin of 6.2 g/dL (reference
range, 12 – 16). A battery of additional hematologic tests show that Anne has β-thalassemia,
intermediate type.
Questions:
1. Describe the types of mutation that cause β-thalassemia. Give specific answers.

2. One type of thalassemia is caused by a nonsense mutation. Codon 17 of the β-globin
chain is changed from UGG to UGA. This change results in the conversion of a codon
for a tryptophan residue to a stop codon. Is it likely that Anne has this mutation in codon
17? Explain your answer.

LABORATORY:
INTRODUCTION
Nucleic acids are the most fundamental constituent of a living cell. They generally serve
as the store houses and carriers of genetic information. There are two types of nucleic acid:
ribonucleic acid (RNA) and deoxyribonucleic acid (DNA). All living organisms contain DNA.
Within the nucleus of every cell are long strings of DNA, the code that holds all the
information needed to make and control every cell within a living organism. DNA, which stands
for deoxyribonucleic acid, resembles a long, spiraling ladder. It consists of just a few kinds of
atoms: carbon, hydrogen, oxygen, nitrogen, and phosphorus. Combinations of these atoms form
the sugar-phosphate backbone of the DNA — the sides of the ladder, in other words.
Other combinations of the atoms form the four bases: thymine (T), adenine (A), cytosine
(C), and guanine (G). These bases are the rungs of the DNA ladder. (It takes two bases to form
a rung — one for each side of the ladder.) A sugar molecule, a base, and a phosphate molecule
group together to make up a nucleotide. Nucleotides are abundant in the cell’s nucleus.
Nucleotides are the units which, when linked sugar to phosphate, make up one side of a DNA
ladder.
During DNA replication, special enzymes move up along the DNA ladder, unzipping the
molecule as it moves along. New nucleotides move in to each side of the unzipped ladder. The
bases on these nucleotides are very particular about what they connect to. When the enzyme
has passed the end of the DNA, two identical molecules of DNA are left behind. Cytosine (C)
will “pair” to guanine (G), and adenine (A) will “pair” to thymine (T). How the bases are arranged
in the DNA is what determines the genetic code.
When the enzyme has passed the end of the DNA, two identical molecules of DNA are
left behind. Each contains one side of the original DNA and one side made of “new” nucleotides.
It is possible that mistakes were made along the way — in other words, that a base pair in one
DNA molecule doesn’t match the corresponding pair in the other molecule. On average, one
mistake may exist in every billion base pairs. That’s the same as typing out the entire
Encyclopedia Britannica five times and typing in a wrong letter only once!
Name: _____________________________ Date: _____________________________

Activity No. 6
CELL DIVISION AND DNA REPLICATION
OBJECTIVES
At The end of the laboratory session, you should be able to

1. Show the replication of DNA before cell division using paper templates for the
components of DNA nucleotides.
2. Explain how DNA replication in higher animals happen.
3. Enumerate the steps of mitosis and meiosis and relate to this to DNA replication

MATERIALS: reference materials, cut-outs of basic subunits of DNA, colors, markers, scissors,
tape or glue, paper and pencil
Scoring rubric
For Rationalization
o 3 – all concepts were mentioned and a clear hypothesis, generalization and conclusion was
o 2 – one or 2 concepts were not mentioned and the hypothesis, generalization and conclusion is
not clear
o 1 – 3 or more concepts were not mentioned and the hypothesis, generalization and conclusion
is not clear
5 points is given for every illustration/documentation made
o 4 – excellent – the illustration shows above average effort on the part of the maker, 1 or 2 parts
o 3 – Good – the illustration shows average effort on the part of the maker, 3 to 4 parts are not
o 2 – Fair – the illustration shows below average effort on the part of the maker, 5 or more parts
labelled
PART 1: ASSIGNMENT
Look for separate illustrations of mitosis and meiosis study them carefully and attach the
illustrations here.

PART 2: DNA REPLICATION
LIFTED FROM: https://www.biologyjunction.com/dnareplication_lab.htm
Procedure:
1. Cut out all of the units needed to make the nucleotides from the handout provided.
2. Color code the Nitrogenous bases, phosphorus, and deoxyribose sugar as follows —
Adenine = red, Guanine = green, Thymine = yellow, Cytosine = blue, Phosphate =
brown, and Deoxyribose = purple.
3. Using the small squares and stars as guides, line up the bases, phosphates and sugars.
4. Now glue the appropriate parts together forming nucleotides.
5. Construct DNA model using the following sequence to form a row from top to bottom –
cytosine (topmost), thymine, guanine, and adenine (bottommost).
6. Let this arrangement represent the left half of your DNA molecule.
7. Complete the right side of the ladder by adding the complementary bases. You will have
to turn them upside down in order to make them fit.
8. Your finished model should look like a ladder.
9. To show replication, separate the left side from the right side, leaving a space of about
6-8 inches.
10. Use the remaining nucleotides to complete the molecule using the left side as the base.
11. Build a second DNA model by adding new nucleotides to the right half of the original
piece of the molecule.
12. Tape the nucleotides together to form 2 complete DNA ladders.
13. Document your work.

Author Janice Friedman

Scoring Rubric
1. Of the 4 bases, which other base does adenine most closely resemble?
2. List the 4 different nucleotides.
3. Which 2 molecules of a nucleotide form the sides of a DNA ladder?
4. If 30% of a DNA molecule is Adenine, what percent is Cytosine?
5. What does the term replication mean?
6. What is another name for adenine and three phosphate molecules attached to it?
7. Describe tests for the following components of DNA
a. Deoxyribose c. Purine
b. Phosphate d. Pyrimidine
8. Describe tests for the following components of RNA
a. Pentose b. Uracil
9. Describe a way to isolate RNA from a sample. Include tests for the products of
hydrolysis
10. What is the “central dogma”, explain
11. Describe ways by which the cell prevents “mistakes” in DNA replication especially during
mitosis

Week no. 5
Submodule no. 5
enzymology

LESSON: Unit 4: Enzymes
TOPICS:
4.1. Definition, Chemical Nature and Importance of Enzymes
4.2. Nomenclature of Enzymes
4.3. Classification of Enzymes
4.3.1. IUPAC Classification 4.3.2. Enzyme Variants
4.3.3. Enzyme Classification in the Blood
4.4. Factors Affecting Enzyme Action
4.5. Enzyme Inhibition
TIME/SCHEDULE: Week no 5
LECTURE: 6 hours
LABORATORY: 3 hours
LEARNING OBJECTIVES
At the end of the learning session, the student should be able to

1. define correctly the different terms encountered in Biochemistry.
2. Describe accurately the nature of enzymes;
3. Correctly comprehend how enzyme kinetics occur;
4. Distinguish correctly the different factors that will influence an enzymatic reaction
5. Perform confidently the assigned activities in the laboratory

LESSON PROPER
INTRODUCTION
Enzymes are proteins that act as catalysts, compounds that increase the rate of
chemical reactions. Enzyme catalysts bind reactants (substrates), convert them to products and
release the products. Although enzymes may be modified during their participation in this
reaction sequence, they return to their original form at the end. In addition to increasing the
speed of reactions, enzymes provide a means for regulating the rate of metabolic pathways in
the body.
ENZYMOLOGY
- It refers to the science of enzymes to the diagnosis and treatment of diseases
ENZYMES
- Substance that catalyzes a given chemical reaction, cellular catalyst
- They are biologic catalysts that causes reactions in the body to take place
- There are over 1,500 enzymes and many of which catalyzes the same reactions
CHARACTERISTICS OF AN ENZYME
- One of the most complicated type of proteins in terms of both structure and functions
- Proteins in nature; easily denatured with varying molecular weight and masses
- Enzymes are amphoteric
- Enzymes operate in high rates
- Turnover number of enzymes is 5x106. Where at this number, these are capable of
converting substrates (which have been made to react with enzymes) into products
- It refers to the science of enzymes to the diagnosis and treatment of diseases
ENZYMES
- Substance that catalyzes a given chemical reaction, cellular catalyst
- They are biologic catalysts that causes reactions in the body to take place
- There are over 1,500 enzymes and many of which catalyzes the same reactions
ACTIVITY:
DEFINE THE FOLLOWING TERMS

1. Coenzyme 5. Prosthetic group
2. Activators 6. Metaloenzyme
3. Holoenzyme 7. Substrate
4. Apoenzyme 8. Product
ENZYME STRUCTURE
- It refers to the molecular/structural arrangement of enzyme molecule

4 Existing Structures
1. Primary structure –
refers to the sequence
of amino acids joined
by peptide bonds
2. Secondary structure
– conformation of the
steric arrangement or
conformation of the
segments of
polypeptide chain
3. Tertiary structure –
arises from the
interactions among
side chains/groups of
the polymer chain;
folded structure of an
enzyme
https://www.slideshare.net/SabahatAli9/protein-structure-levels
4. Quaternary Structure – refers to the relationship between subunits
- Subunits perform specific functions
o The secondary and tertiary structures are the most important configurations of
the enzyme because these structures are responsible for the characteristics such
as coiling and folding resulting to conformational structure
Active Site
- Refers to the area or portion of an enzyme in which the substrate is attached with the
enzyme molecule
- It is where the transformation of the substrate takes place
https://www.philpoteducation.com/mod/book/view.php?id=782&chapterid=1127#/
Classification of Enzymes
- Based on catalytic activity of an enzyme

- The International Union of Biochemistry (IUB) enzyme commission categorized all
enzymes into 6 classes
o Oxidoreductases
o Transferases
o Hydrolases
o Lyases
o Isomerases
o Ligases
OXIDOREDUCTASES
- Enzymes catalyzing oxidation and reduction reactions

o Reduction – addition of hydrogen to a double bond
o Oxidation – removal of hydrogen to cleave a double bond
- Older names are dehydrogenases and oxidases
- Assayed for investigation of gives information regarding heart attacks and liver problems
- Examples of oxidoreductases
- Oxidases
o Glucose oxidase
o Cytochrome oxidase
- Dehydrogenase
o LDH
o Iditol DH
o MDH
o Isocitrate DH
o Glucose-6-PO4 DH
o Hydroxybutyrate DH
- Oxidoreductases acts on
o Secondary alcohols
o Ketones
o Alkenes
o Primary amines
o Secondary amines
o NADH, NADPH
• Example:
• Pyruvate + NADH+ + LDH = lactate + NAD
TRANSFERASES
- Enzymes that move and interact group of atoms from one molecule to another (amine or
PO4 group)
- Gives important information about liver damage
o Examples of transferases
o Aspartic aminotransferase (AST)
o Serum glutamate oxaloacetate transaminase (SGOT)
o Alanine aminotransferase (ALT)
o Serum glutamate pyruvate transaminase (SGPT)
o Creatine kinase
o Gamma glutamyl transferase
o Ornithine carbamyl transferase

• Example:
• ATP + creatine + CK = ADP + creatine phosphate
HYDROLASES
- Enzymes involved in the splitting of molecules with water as a part of the reaction
process
- 3 groups of hydrolases
Esterases Peptidases Glycosidases

ACP Leucine aminopeptidase Amylases
ALP Trypsin Amylo-6-glucosidase
Cholinesterase Pepsin Glucoside
Lipase Galactosidase
- Example: amylase cleaves the C-O-C bonds of starch
LYASES
- Enzymes responsible for splitting molecules (lysis)

- Bonds broken are
o C-C bond
o C-O bond
o C-N bond
o Others
- Assayed in disorders of skeletal muscles
- Examples
o Aldolase
o Glutamate decarboxylase
o Tryptophan decarboxylase
ISOMERASES
- Enzymes responsible for the conversion of one isomer to another

- It involves the transformation from
o Cis-trans
o L-D forms
o Aldehyde to ketone
- All reactions are reversible
- No clinical importance
- Examples
o Glucose PO4 isomerase
o Ribose PO4 isomerase
LIGASES
- Enzymes causing bond formation between two molecules to form a longer molecule
- Example

o Ligase enzyme
o Amino-acyl-tRNA synthetase – activates amino acids before incorporation to the
polypeptide chain
ENZYME NOMENCLATURE
1. Each enzyme was designated according to the reaction it catalyzes

Example: oxidation of glucose – glucose oxidase
2. It is customary to identify enzymes by adding the suffix “ase” to the group upon which
the enzyme acts
Example: Urea – urease
Uric – uricase
Phosphate esters – phosphatase
Lipid – lipid esters
Lactose – lactase
Starch – amylase
3. Naming of enzymes by the use of their empirical names
Example: trypsin, pepsin
4. Standard system of identifying enzymes
- Formulated by IUB and IUPAC
o Systematic name
▪ Describe the nature of the reactions catalyzed
▪ Unique numerical code designation (consists of 4 members separated by
periods)
▪ Prefixed with letter E.C. (Enzyme Commission)
• E.g. E.C. 3.1.3.1. – ALP
E.C. 3.1.3.2 – ACP
o 1st digit – denotes the class of the enzyme

o 2nd digit – denotes the sub-class of the enzyme
o 3rd digit – denotes the subsub-class
o 4th digit – specific serial number given to each enzyme
within its subsub-class
▪ This approach removes all ambiguity about the
enzyme’s identity
o Trivial names
▪ A.K.A. non-specific , practical name, working name
▪ Identical to systematic name and often a simplification of it
▪ Suitable for everyday use
▪ Uses acronyms and abbreviations
• Orthophosphoric monoester phosphorylase - ALP
ENZYME VARIANT
- These are the several distinct forms of enzymes
- Important in the diagnosis of specificity

A. ISOENZYMES
- The multichained enzymes
- Enzymes of similar activity typically appearing in specific tissue, organs and cell
organelles of organisms and the same species
- Specifically located in tissues
- The site of a specific isoenzyme is found and restricted to only one tissue
o Example: Lactate dehydrogenase: A and M
▪ Contains H and M subunits (polypeptide chains)
▪ The subunits interact together for a reaction to occur
• H4 – LD1 – heart, RBC, and renal tissues
• H3M – LD2 – heart, RBC and renal tissues
• H2M2 – LD3 – lungs, lymphocytes, spleen, pancreas
• HM3 – LD4 – liver, skeletal muscle
• M4 – LD5 – liver, skeletal muscle
- Characteristics
o Charged molecules – shown in electrophoresis
o Mobility in Ion Exchange Resin
o Response to activation and inactivation process
▪ E.g. ACP (RBC) – not inhibited by 2% formaldehyde
ACP (prostate) – inhibited by 2% formaldehyde
o Relative substrate specificity
▪ E.g. ACP (RBC)- less sensitive to α-naphthyl PO4 substrate
o Response to inhibitors
B. HETEROENZYMES
- Same enzymatic activity, which are specie specific for different biological species
e.g. LDH of rabbits and humans
C. ALLOENZYMES
- Genetically transmitted enzymes
- Important in defining the biochemical characteristics of individuals
- Present only in some selected individuals of the same species
- Practical value in forensic medicine and genetics
ORIGIN OF PLASMA ENZYMES: CLASSIFICATION OF ENZYMES IN THE BLOOD

A. Plasma Specific Enzymes
- Generally secreted by liver into the plasma
- Enzymes exert function in plasma
- Examples
o Blood clotting mechanisms
o Complement system and those connected with fibrinolysis
o Thrombin, factor XII (Hageman factor), Factor X (Stuart factor)
o Precursors (plasminogen activators)

o Serine protease procoagulants
B. Non-plasma specific enzymes

- Enzymes that do not have specific function in plasma
- The plasma lacks activators or coenzymes which are necessary for enzyme activity
- 2 classes
o Enzyme of secretion
▪ These are normally secreted enzymes like AMS
▪ Enzymes secreted in plasma at a very high rate but are rapidly disposed
off to normal excretory channels thus concentration in plasma is
maintained at low and constant level
o Enzymes associated with cellular metabolism
▪ Enzymes that carry out their functions within the cells in which they are
formed
• E.g. CK in heart and ACP in prostate gland
Enzymes based on distribution

A. Unilocular Enzymes
- These are enzymes found only in the cell sap
- Found only in one location
B. Bilocular enzyme
- These are enzymes that are found in the mitochondria and cell sap
Enzyme Kinetics
E+S ES P+E
Where:
E = enzyme that represents a true catalyst
= not changed in the reaction
S = substrate upon which the enzyme acts
P = the reacted substrate
= product that represents a changed molecular species of substrate
ES = the postulated enzyme – substrate complex
I. Factors Affecting the Binding
A. Energy – this refers to the activation energy

B. Molecular compatibility – E + S = commonness and sameness between
enzyme and substrate
C. Space availability – refers to the number of enzymes or substrates that can be
reacted
D. Specificity – refers to the particular enzyme catalyzing a specific substrate

II. Factors Affecting E and S Combination
A. Lock and Key – refers to the active site being complementary in shape and size
of the substrate
B. Induced Fit model – refers to the enzyme changing in shape during binding with
substrate. After binding, the shape of the enzyme complements with the binding
site
III. Factors influencing Enzymatic Reaction
A. Substrate concentration
a. direct relationship
b. An increase in the concentration of substrate results to an increase in the
rate of enzyme activity
c. No free binding site
B. Time
a. This refers to the condition when the enzymatic reaction is allowed to take
place
b. If the catalytic activity of an enzyme on the substrate is fast, it results to a
shorter enzymatic reaction time thereby the enzyme is freed and can act
again on the remaining substrate
2 Phases of Enzyme Reaction

A. First Order Kinetics
• Enzyme concentration is fixed and the substrate concentration is
varied
• The rate of reaction is almost directly proportional to substrate
concentration at low values (lower substrate concentration than
enzyme)
• The rate of reaction dependent on substrate concentration and on
enzyme concentration
B. Zero Order Kinetics
• The rate depends on the enzyme concentration and independent
of substrate concentration

• An increase in the enzyme concentration results to an increase in
the catalytic activity of the enzyme
https://biolympiads.com/michaelis-menten-equation/
C. Enzyme Concentration
a. Direct relationship
b. An increase in the enzyme concentration result to an increase in the
catalytic activity of the enzyme
D. Temperature
a. Optimum temperature – the point at which the enzyme molecule is
capable of catalyzing the substrate with regards to temperature
requirement
i. An increase of 10C in temperature results in doubling the rate of
enzyme activity
ii. 30-37C or 37-40C – optimum temperature
iii. 60-70C – enzyme undergoes inactivation and denaturation
iv. Further increase in temperature leads not to further increase in the
rate but loss of enzymatic assay
v. Some are stored at refrigeration temperature (2-8C)
b. Increased rate in
i. Increasing movement of molecule
ii. Increasing rate at which internal cellular collision
c. Q10 value – refers to the increased reaxtion rate for every 10C increase
(rate of reaction is doubled)
E. pH – Hydrogen ion concentration

a. optimum pH – the point at which the reaction rate is greatest
i. at pH 7-8, many enzymes show maximum activity
ii. pepsin is active at 1.5 and ALP at 10.5
F. Activators
a. Required for full enzyme activity

b. These metal activators or inorganic entities help bind the substrate tp the
active site by forming ionic bridges
c. These help substrate in orientation so it is attached to the protein at the
proper point nd in the correct configuration
Examples:
Divalent Cations Monovalent cations Monovalent cations

- Fe++ - Cl- K+
- Ca++ - Br- Na+
- Zn++ - NO3-
G. Inhibitors
a. Substances that decrease the rate of enzyme reaction if these are
present in the reaction system
b. Functions
i. Binds to the active site, blocking the access of the substrate to the
enzyme (competitive inhibition)
ii. Binds elsewhere on the enzyme causing change in shape that
interferes with substrate binding (non-competitive)
iii. Uncompetitive – inhibitors bind with the ES completely; no
product formed
https://www.youtube.com/watch?v=UostSikM888

c. Types of inhibition
i. Reversible inhibition
1. When inhibitors are possible removed from the system and
enzyme is fully restored
2. Physical separation processes that remove inhibitors
a. Dialysis
b. Gel filtration
ii. Irreversible inhibition
1. When inhibitors combine covalently with the enzyme
2. Physical methods are ineffective in separating inhibitors
from enzymes
d. Examples of inhibitors
i. Excess substrate
1. Causes competition between substrate molecules for a
single binding site
ii. Product of Reaction
1. A product of reaction may itself be an inhibitor of the
forward reaction
iii. ES does not break to give products
iv. Chemical substances
1. Fluoride
2. Heparin
3. Oxalate
4. Cupric
5. Tartrate
H. Coenzyme concentration
a. Many enzymes require a coenzyme of some sort for the reaction to
proceed (vitamins, CoE, A or B, NAD)
b. These must be present at the proper concentration for many enzyme
reactions to take place
I. Prosthetic Group
a. These are essentially coenzymes that firmly bind with enzymes
Enzyme Denaturation
- It refers to the disruption of the structure of enzyme molecule (3-dimensional structure)
that leads to the loss of enzyme activity
- Denaturing Conditions
o Elevated temperature
▪ Weakens the stabilizing bonds
▪ Above 60C
o Extremes of pH
▪ Causes unfolding of enzyme molecule
o Radiation
o Frothing

o Strong salt solution
o Mechanical trauma
o Time of exposure
o Chemicals
- Denaturation may be reversed if the reaction process has not gone too far and if the
denaturing agent is removed
Types of Enzyme Specificity
Activity: Define the following and indicate at least two specific examples of enzymes
1. Absolute specificity
2. Group Specificity
3. Stereoisomeric
Units of Measurements – proposed by the IUPAC and IUB
A. International Unit (IU)

- Defined as the amount of enzyme which catalyzes the conversion of 1umol of substrate
in 1 minute
- Reported in IU/liter of sample
B. Katal
- A unit of enzyme activity which converts 1 mol of substrate per second
- Has not yet received universal acceptance
Activity: Define the following units of measurements for enzymes:
A. Bondansky -
B. Gutman-Gutman -
C. BLB -
Enzyme Regulation
1. Feedback control – the concentration of the product influences the rate of reaction.
2. Allosterism – an interaction takes place at a position other than the active site but
affects the active site, either positively or negatively.
3. Some enzymes (proenzymes or zymogens) must be activated by removing a small
portion of the polypeptide chains.
4. Zymogens are enzymes produced as inactive proteins.
5. Enzyme activities are also regulated by isoenzymes (isozymes), enzymes that catalyze
the same reactions but vary slightly in structure.
THE WATER-SOLUBLE VITAMINS
1. Thiamine (B1) – coenzyme: thiamine pyrophosphate

a. Fresh vegetables, husks of cereal grains, liver and other organ meats
b. A vitamin deficiency disease seen particularly among those who eat large quantities of
polished rice. Symptoms include weakness, neuritis, and in severe cases, mental
disturbance and heart failure.
c. It can be cured by giving large doses of vitamin B.

2. Riboflavin (B2) – coenzyme: flavin adenine dinucleotide (FAD) and flavin mononucleotide
a. Milk and meat products
b. Dermatitis; glossitis; cataracts
3. Niacin (nicotinic acid B3) – coenzyme: nicotinamide adenine dinucleotide (NADH)
a. Liver, lean meats, cereals, and legumes
b. A vitamin deficiency disease characterized by a rash on exposed body surfaces,
diarrhea, ulcerations in the mouth and mental disorientation (pellagra).
c. It may be seen among alcoholics who drink rather than eat.
4. Pyridoxine (B6) – coenzyme: pyridoxal phosphate
a. Meats, cereals, lentils (legumes)
b. Dermatitis, fatigue, anemia, irritability
5. Cobalamin (B12) – coenzyme: nicotinamide adenine dinucleotide (NADH)
a. Milk and meat products
b. Pernicious anemia, malformed RBCs, mental disorders
c. The administration of vitamin B12, folic acid, liver and stomach extracts have been
found effective in control of this condition.
6. Ascorbic acid
a. Plants, fruits, and vegetables
b. Scurvy – bleeding gums, slow-healing wounds,mental disorders
7. Biotin – coenzyme: biocytin
a. Yeasts, meats, dairy products, grains, fruits and vegetables, intestinal bacteria
b. Scale skin, fatigue and muscle pains, anemia, nausea, weakness; depression
8. Folic acid – coenzyme: tetrahydrofolic acid
a. Organ meats, fresh green vegetables
b. Abnormal red and white blood cells (anemia); intestinal tract disturbances
9. Pantothenic acid – coenzyme: coenzyme A
a. Most foods, especially liver, meat, cereals, milk, fresh vegetables
b. Vomiting, abdominal distress, insomnia
Most Common Disturbances
1. Kwashiorkor – disease resulting from a deficiency of dietary protein relative to caloric intake
(protein-energy malnutrition).
a. The disorder occurs most commonly in young children in developing countries when
mothers’ breast milk no longer provides enough protein, and other protein-rich foods are no
given in sufficient quantity.
b. The number of calories in the diet of corn meal is sufficient, but corn is deficient in
lysine and tryptophan. As a result, a child fails to grow, becomes lethargic, and has an enlarged
abdomen from hypoprotein edema due to the inability to properly build proteins.
2. Marasmus – results from slow starvation caused by deficiency not only of protein but also of
calories and other nutrients.
3. Anemia – blood condition involving an abnormal reduction in the number of RBCs or in their
hemoglobin content.
4. Gout – caused by faulty metabolism of uric acid produced in the body by breakdown of
proteins.
▪ Factors: diet rich in malt liquors, wines, and certain types of protein and about 95% are
men.

5. Phenylketonuria (PKU) – this is due to the absence of the enzyme phenylalanine hydrolase
which converts phenylalanine to tyrosine. Phenylalanine is instead converted to phenylpyruvic
acid which impairs normal development of the child’s brain leading to mental retardation.
6. Albinism – this is due to the absence of tyrosinase, which is necessary for the formation of
melanin.
7. Alkaptonuria – this is due to the absence of the homogentisic acid oxygenase (homogentisic
acid → acetate and fumarate).
▪ A disturbance of metabolism in which the skin of the face, the whites of the eyes, and
other tissues such as muscle and cartilage become discolored brown. (The urine is also
a dark, brownish color.)
8. Tyrosinosis – this is due to the absence of hydroxyphenylpyruvic acid oxygenase which

converts hyroxyphenylpyuvic acid to homogentisic acid.
a. Infants exhibit diarrhea, vomiting, and a “cabbage-like” odor and fail to thrive.
b. Without treatment, death from liver failure ensues 6-8 months.
c. Treatment: diet low in tyrosine, phenylalanine, and methionine.
1. ____________________________ It refers to the science of enzymes to the diagnosis
and treatment of diseases
2. ____________________________They are biologic catalysts that causes reactions in
the body to take place
3. ____________________________Term for the inactive state of the enzyme
4. ____________________________ A substance often derived from a vitamin, and may
be associated with different enzymes
5. ____________________________ A term that is used to describe inorganic ionic
cofactors
6. ____________________________
7. ____________________________ What are the two components making up
holoenzymes
8. ____________________________ What do you call a coenzyme that cannot be
removed from its attachment with an enzyme using dialysis
9. ____________________________ The type of bond that holds the tertiary structure of
enzymes.
10. ____________________________ Part of the enzyme where inhibitors attach during
non-competitive inhibition
11. ____________________________ Which group of enzymes are assayed for
investigation or gives information regarding heart attacks and liver problems
12. ____________________________Which group of enzymes that move and interact
group of atoms from one molecule to another (amine or PO4 group)
13. ____________________________ A substance needed by hydrolases in order to split
molecules
14. ____________________________ In the systematic enzyme nomenclature what does
the first digit denote?

15. ____________________________ What do you call enzymes of similar activity typically
appearing in specific tissue, organs and cell organelles of organisms and the same
species?
16. ____________________________ Which enzyme variant of lactate dehydrogenase is
found in the lungs, lymphocytes, spleen, pancreas?
17. ____________________________ location of the ACP that is inhibited by 2%
formaldehyde
18. ____________________________ location of the ACP that is less sensitive to α-
19. ____________________________ These are genetically transmitted enzymes,
important in defining the biochemical characteristics of individuals
For numbers 20 – 23, classify the following as being (P) plasma specific and (N) non-
plasma specific enzymes.
20. ____________________________ Hageman Factor
21. ____________________________ Serine protease procoagulant
22. ____________________________ ACP
23. ____________________________ CK
24. ____________________________What do you call enzymes secreted in plasma at a
very high rate but are rapidly disposed of to normal excretory channels thus
concentration in plasma is maintained at low and constant level
25. ____________________________ An enzyme model that refers to the active site being
complementary in shape & size to the substrate
26. ____________________________ a graph that shows the relationship of the reaction
velocity to the substrate concentration
27. ____________________________ These are substances that help orient the substrate
so it is attached to the enzyme in the correct configuration
28. ____________________________ A type of enzyme that acts only on a specific isomer
29. ____________________________ Defined as the quantity of enzyme that will catalyze
the reaction of 1 µmol of substrate per minute
30. ____________________________ A unit of enzyme activity w/c converts 1 mol of
substrate per second
ASSIGNMENT:
1. Using comic strips, describe your understanding on the different functions of inhibitors.
2. What is the role of vitamins in enzyme activity? Cite specific examples.
3. Answer the activities found in this unit
Scoring Rubrik
4 3 2 1
unsuccessful
accuracy

correctly cited
slows readability
Case:
Sam Miguel is a 44-year-old man who has been an alcoholic for the past 5 years, had a
markedly diminished appetite for food. One weekend he became unusually irritable and
confused after drinking two fifths of scotch and eating very little. His landlady convinced him to
visit the doctor. Physical examination indicated a heat rate of 104 beats per minute. His blood
pressure was slightly low and he was in early congestive heart failure. He was poorly oriented
to time, place and person.
After being released from the hospital, he continued to drink. One night he arrived at a
friend’s house at 7:00 pm. Between his arrival and 11:00 pm, he drank four beers and five
martinis (for a total ethanol consumption of 9.5 oz). His friends encouraged him to stay an
additional hour and drink coffee to sober up. Nevertheless, he ran his off the road on his way
home; He was taken to the emergency room of the local hospital and arrested for driving under
the influence of alcohol. His blood alcohol concentration at the time of his arrest was 240 mg/dL,
compared to the legal limit of ethanol for driving of 80 mg/Dl.
Questions:
1. Sam Miguel has developed thiamine deficiency. What is the effect of alcohol in the
transportation of thiamine through the intestinal mucosal cells that eventually caused the
dysfunctions that occurred in his central and peripheral nervous system as well as the
cardiovascular system? Include the enzymes affected by this type of deficiency and the
name of the disease that results from thiamine deficiency.
2. What enzyme in the liver metabolizes ethanol from alcoholic drinks? Show the equation
on how this enzyme acts on ethanol and how the product may cause liver damage.
3. What is the rate at which ethanol is cleared from the blood? Would the one hour being
asked by his friend to stay to sober up suffice? If not, how many hours would it take to
clear the ethanol from Sam’s blood?

LABORATORY:
INTRODUCTION
Enzymes are of fundamental importance in many of the chemical reactions, which takes place
in living organisms. When digestion occurs, enzymes released into the mouth, stomach and
intestines catalyze reactions, which results in the breakdown of large foodstuffs into building
block molecules.
Enzymes are protein molecules and proteins have a characteristic three-dimensional structure
called conformation. Any environmental conditions that destroy the protein structure will also
destroy tis enzyme activity.
The molecules upon which enzymes acts upon are called substrates. The part in the enzyme
too which the substrate attach to is called the active site. The active site is where the
transformation of the substrate takes place to produce the product.
Name: _____________________________ Date: _____________________________

Activity No. 7
ENZYMES AND DIGESTION
OBJECTIVES:
At the end of the laboratory session, you should be able to:
A. demonstrate accurately how lipase digest fats in milk.
B. enumerate correctly the products of the hydrolysis of fats.
C. state appreciatively the importance of enzymes in digestion.
MATERIALS: Use reference materials and our previous lectures to help you arrive at the
correct answer.
Marker pen, test tube rack, syringe without the needle, beakers (100 mL and 250 mL),
thermometer, test tube, glass rod, stop watch, water bath, Bunsen burner
CHEMICALS: Full cream milk, phenolphthalein, 5% lipase, 0.05M sodium carbonate, ice
PROCEDURES AND OBSERVATION
Scoring rubric
For Rationalization
not clear

is not clear
Phenolphthalein is an indicator that is pink in alkaline solutions of about pH10. When the
pH drops below pH 8.3 phenolphthalein goes colorless. A solution of milk and lipase is made
alkaline by the addition of sodium carbonate and phenolphthalein will change from pink to
colorless as the fat in milk is broken down.
PROCEDURE:
You will not perform the following procedure but will use it to answer the questions
found below.
Preparation
1. Prepare a water bath with the following temperatures: 0C (use ice bath), 37C and 60C.
2. Place 2.5 mL of lipase in 3 separate test tube and put this in the three water baths
prepared in procedure 1
Investigation
1. Label three test tubes with the temperature to be investigated.

2. Add five drops of phenolphthalein to the test tubes
3. Place 5mL of milk in each of the test tubes plus 7 mL of sodium carbonate solution
4. Place a thermometer in each of the test tube and place them in the corresponding water
bath
• You may remove the thermometer once the contents of the test tube reaches the
same temperature as the water bath.
• At this point, there are two test tubes in your water bath, one containing the
lipase, the other containing the milk solution.
5. Using a clean pipet or syringe, measure 1 mL of the lipase from the 0C water bath and
add this to the test tube containing milk in the same water bath. Start the timer.
6. Do the same for the 37C and 60C
7. Using applicator sticks, stir the milk solution without removing them from the water bath
until the pink color is discharged.
8. Note the time it takes for the three test tubes to change in color.
Illustration:
- Draw the procedures above in a short bond paper with one-inch margin in all sides and
label all the steps properly.
- Scan your illustration and pass together with this manual.
Using the procedure above, answer the following questions using your reference
materials.
1. Write the complete equation for the hydrolysis of fats.

2. When fats are hydrolyzed, what happens to the pH of the resulting solution?
3. After the hydrolysis of fats, which product will change the color of phenolphthalein from
pink to colorless? Explain your answer.
4. In which temperature do you think will yield the fastest result? Reason out your answer.
5. What is the role of bile acids in the digestion of fats?
6. If starch were to be hydrolyzed using amylase, what are the products that will be
produced? Answer by writing the complete word equation.
7. What simple test can you use to detect the presence of the end product of the hydrolysis
of starch using amylase? Include the positive result.
8. Starch is actually hydrolyzed in stages; enumerate the stages in chronological order.

Scoring Rubric
1. How is temperature related to enzyme activity?

2. What is Q10 value in relation to enzyme activity?
3. What do you think is the influence of temperature on the protein structure of the enzyme
to affect its activity?
4. How is pH related to enzyme activity?
5. What do you think is the influence of pH on the protein structure of the enzyme to affect
its activity?
6. Explain the importance of enzymes in digestion.

WEEK NO. 6
SUBMODULE NO. 6
THE
BIOCHEMISTRY OF
DIGESTION

LESSON: Biochemistry of Digestion
TOPICS:
5.1 Definition
5.2. Factors Affecting Digestion
5.3. Phases of Digestion
5.3.1 Salivary Digestion
5.3.2 Gastric Digestion
5.3.3. Intestinal Digestion
5.3.4. Pancreatic Juice
5.3.5. Intestinal Juice
5.3.6. Bile
5.4. Chemical changes in the large intestines and feces
5.4.1. Overview
5.4.2. Fermentation
5.4.3. Putrefaction
5.4.4. Deamination
5.4.5. Decarboxylation
5.5. Detoxification
5.6 Chemical Composition of Feces
LECTURE: 5 hours
LABORATORY: 4 hours
At the end of the learning session, the student will be able to:
1. Define correctly the different terms encountered in this unit.
2. Discuss completely the digestion of complex biomolecules in the body.

LESSON PROPER:
Introduction:
Digestion is an activity that is done as part of daily living. All living organisms, primarily
animals and humans, entail digestion to sustain daily activities and essential needs. Digestion, in
the context of biochemistry, requires the presence of enzymes, co-enzymes and cofactors. This
process when successful will result in the production of building blocks of the different
biomolecules which are necessary in performing body building, regulatory, and energy-giving
functions. A disruption in this process will impact the health status of an individual.
Lesson 1. Definition and Form of Digestion
DIGESTION is referred to as the disintegration of the naturally occurring foodstuff into

absorbable forms. It includes the hydrolysis of starch into monosaccharides, proteins to amino
acids, and fats into fatty acids and glycerol.
• FACTORS IN DIGESTION
There are two factors in digestion, primary and secondary factors. Primary factor includes
the different enzymes found in the juices of the different portions of the digestive tract. These are
necessary in the conversion of complex molecules to simple and absorbable forms. Secondary
factors, on the other hand, include the following:
- Cooking ruptures walls of starch granules, softening of connective tissue (make
them accessible for e action). It also improves the taste and appearance of food
which in turn stimulate secretion of digestive enzymes
- Mastication breaks down the food and increases surface area for enzymes to act.
It is a mechanical stimulus for the flow of saliva and digestive juices.
- movements of the stomach and intestines or what is known as peristalsis.
- absorption from the intestines,
- autodigestion such as in the ripening of fruits.
The digestion of food undergoes several processes in different parts of the digestive
system. This is referred to as the physical movement and breakdown of food which includes:
1. Swallowing
a. Buccal stage
i. Bolus is pushed toward the pharynx
ii. Voluntary movement
b. Pharyngeal and Esophageal stage
i. Bolus (masticated food with saliva) to pharynx to esophagus to stomach
ii. Involuntary movement
2. Peristalsis in the stomach
a. Mixes contents and forces chime thru the pylorus
b. Three waves travel down the stomach one at a time
i. Each beginning every 20 seconds near the midpoint of the stomach,
lasting about one minute
c. The rate of emptying of the stomach is determined largely by the strength of
contraction
d. Feedback from the duodenum regulates gastric emptying
e. 2 mechanisms that inhibit gastric motility
i. Enterogastric reflex – neuronal
ii. Enterogastrone – hormonal

• Chyme – partially digested food with gastric juices
3. Contraction of the small intestines
a. Segmenting – rhythmic contractions along a section dividing it into segments
i. Primarily a mixing action
b. Gastroileal reflex – the increase in ileal peristalsis and frequency of opening of
the ileocecal valve due to the ingestion of food
4. Contractions of the large intestines
a. Haustra – simultaneous contraction of circular and longitudinal muscle
b. Gastrocolic reflex – infrequent (twice a day) mass movement transferring
contents from proximal to distal colon and into the rectum
i. Occurs shortly after meal
5. Defecation Reflex
a. Distention of rectum triggers intense peristaltic contractions of colon and rectum
and relaxation or internal anal sphincter
b. Reflex preceded by voluntary relaxation of external sphincter and compression of
abdominal contents
1. Compare the following using Venn diagram.
Primary factor Secondary factor
2. Discuss how primary and secondary factors affect digestion?

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
Lesson 2. Different Stages of Digestion
CHEMICAL BREAKDOWN OF FOOD

A. MOUTH
Mechanical digestion in the mouth

• Chewing or mastication
• Food manipulated by tongue, ground by
teeth, and mixed with saliva
• Forms bolus
Chemical digestion in the mouth
• hydrolysis of starch by salivary amylase
(ptyalin)
• takes place in the buccal cavity and to a
certain extent in the fundic end of the
stomach
• Starch & glycogen → maltose
Saliva
o Colorless slightly viscid, opalescent
fluid which is a mixture of secretions
of the three pairs of salivary glands
▪ Parotid gland – section is
watery and rich in ptyalin
▪ Sublingual gland – secretion is
more viscid containing mucin
and poorer in ptyalin
o Average secretion per day is 1,500 ml
o 99.42 % water
o 0.58 % solid
o 2/3 = org. matter (mucin, ptyalin,
urea, glucose, lactic acid…)
o 1/3 = inorg. Salts (Cl-, HCO3- of Na, K,
Ca, SO4- and PO4- )
o pH: 7.0 – 7.3 (active stimulation)
o 6.4 – 6.9 (resting saliva)
o Function: moisten and reduce the
foods into a consistency suitable for
swallowing
Factors influencing the secretion of saliva

1. Psychic
2. Chemical sight, smell and taste of food
3. Mechanical
Salivary amylase
- Also known as ptyalin
- Endoamylase which acts only on the alpha 1,4-glycosidic linkages
- Splits starch into the disaccharide maltose

- activators: Cl- & Br -; I- & NO3
- Unstable below pH 4 – 5
- Inactivated by pepsin
B. STOMACH
Gastric Digestion
- Mainly concerned with the digestion of proteins thru the action of the enzyme pepsin and
HCl
- Pepsin – initiates protein digestion producing proteoses, peptones and polypeptides
Pepsinogen HCl (acid) Pepsin

(Chief cell) (Parietal cell)
3 Phases in Gastric Digestions

1. Psychic phase
a. Stimulation of gastric secretion due to the smell, sight, taste and thought of food
b. Gastrin – secreted by G cells in the pyloric region of the stomach, it stimulates
the secretion of an acid rich gastric juice
2. Gastric Phase
a. Initiated by food in the stomach
b. Secretagogue – present in certain food which stimulates the secretion of gastrin
(hormone from the pyloric mucosa)
3. Intestinal phase
a. Initiated by digestive products in the upper small intestine
b. Secretin – a hormone released by the duodenum acting on the stomach, it
stimulates the secretion of gastric juice, pancreatic juice, intestinal juice and bile
Gastric Juice
- Average secretion per day is 2-3 liters
- Contributed by three types of cells from the gastric glands
o Parietal cells – secrete HCl (0.17N), pH 0.87
o Chief cells – secrete pepsin
o Mucosal cells – secrete mucin
Composition
• 99.4 % water
• 0.6 % solid
• organic constituents: mucin, pepsin, & small amount of lipase
•
Inorganic Constituents: HCl, KCl & PO4-
Pepsin
• principal digestive constituent of the gastric juice
• pepsinogen – activated by HCl; Inactivated at neutral pH or alkaline pH
• Peptide bonds pepsin di or tripeptide/shorter)
• autocatalytic
• Optimum activity: pH 1.5 – 2.5
• An Endopeptidase that acts on peptide bonds
Mucin
• not digested by pepsin
• Protects the mucous membrane of the stomach
• Buffers the HCl

• Promotes absorption of Vit. B12
Renin – milk –curdling enzyme found in the gastric juice of newborn infants
- The gastric juice on infants contain little or no pepsin but high concentration of HCl
Inhibitors of Gastric Secretion

Enterogastrone – produced from the presence of fats in the intestines which inhibits gastric
secretion and motility
Urogastrone – found in urine: similar to entrogastrone
Acids inhibitors
• 0.03 N HCl – slows secretion
• 0.1 N HCl – inhibits parietal cells
Digestion
Functions of HCl
1. Provide suitable pH for protein digestion by pepsin
2. Exerts preliminary swelling, denaturing and hydrolyzing effect on proteins
3. Activates pepsinogen
4. Facilitates absorption of iron
5. Causes hydrolysis of disaccharides
6. Stimulates secretion of secretin
7. Exerts germicidal action
Pathological Variation of HCl

▪ Achloridia:
- absence of free HCl
- seen in Pernicious anemia
▪ Achylia gastrica
- partial or complete absence of gastric juice
- indicates degeneration of the gastric glands
▪ Hypochloridia:
- suggestive of chronic gastritis or gastric carcinoma
▪ Hyperchloridia:
- suggestive of peptic ulcer, cholecystitis.
Lesson 3. Intestinal Digestion
C. INTESTINES
The last site of digestion and the site of absorption is the small intestines. Most enzymes
are present in this particular site thus, chemical digestion is at the most. Take note however,
that in intestinal digestion, it does not focus on the small intestines alone but also includes the
pancreas and the bile.

INTESTINAL DIGESTION
Pancreatic Juice
- Secreted by the acinar cells of the pancreas (500ml/day)
- Clear, watery solution made up of 98.7% water and 1.3% solids
- It is the most alkaline of the normal body fluids (pH 7.5 – 8.2) due to its bicarbonate content
o Organic constituents (1/3)
▪ Trypsin, chymotrypsin (proteolytic enzymes)
▪ Steapsin (lipolytic)
▪ Diastase (amylolytic)
▪ Nucleolytic enzymes
o Inorganic constituents (2/3)

▪ Sodium
▪ Potassium
▪ Calcium

▪ Bicarbonate
▪ Chlorides
▪ Phosphates
Regulation of Pancreatic Secretion

1. Vagal Stimulation
a. Secretion is rich in enzymes but less volume
b. Occurs during cephalic and gastric secretion
2. Hormonal stimulation
a. Secretin – stimulates secretion of a fluid of high volume and bicarbonate content
b. Pancreozymin – stimulates enzyme secretion
Enzymes in the Pancreatic Juice

1. Trypsin
- Protein splitting enzyme and an endopeptidase
- Secreted as trypsinogen and initially activated as trypsin by enterokinase from succus
entericus
- Acts upon peptide linkages
- Splits proteins and products of pepsin digestion into peptides
2. Chymotrypsin
- Endopeptidase acting on peptide bonds involving phenylalanine, tyrosine and tryptophan
- Activated by trypsin
- Powerful milk-curdling effect
o Peptidases
▪ All intestinal enzymes except two
▪ Split peptides into amino acids
3. Carboxypeptidase
- Zinc-containing enzyme
- Exopeptidase attacking the peptide linkage nearest the free hydroxyl group
4. Pancreatic amylase
- Also known as diastase and amylopepsin
- Converts starch and glycogen into maltose which is immediately acted upon by the
enzyme maltase
5. Pancreatic Lipase
- Also known as steapsin
- It exhibits weak lipolytic effect
- Action is potentiated by bile acids, calcium and certain amino acids
- It splits fats into monoglycerides, fatty acids and glycerol
6. Nucleodepolymerase
- Ribonucleases and deoxyribonucleases
- They hydrolyze the corresponding nucleic acid into mononucleotides
Intestinal Juice (Succus Intericus)

- Combined secretion of the intestinal glands at different portions: duodenum, jejunum,
ileum
- Viscous and turbid fluid due to mucin and desquamated epithelial cells
- Enterocrinin – hormone secreted by the intestinal mucosa and stimulates the mucosal
glands to increase volume of fluid and the content of enzyme

Enzymes in Intestinal Juice
1. Carbohydrases
a. Maltase
b. Lactase
c. Sucrose (invertase)
2. Peptidases
a. Aminopeptidase – exopeptidase, attacks the peptide linkage of the terminal
amino acid containing free amine group which requires Mg ++ or Mn++
b. Tripeptidases and Dipeptidases – split tripeptides and dipeptides into their
constituent amino acids which require Co++ and Mn++
If you try to look at this figure,
it shows the function of both
the aminopeptidase and
carboxypeptidase. Since both
are exopeptidases, they act
on the peptide linkage of the
terminal amino acid (the one
in circle) which is the amine
group for aminopeptidase and
carboxyl group for
carboxypeptidase.
3. Nucleotidases (Nucleophosphatases) – split the nucleotides into phosphoric acid

nucleosides
Nucleotides Nucleotidase Nucleoside + Phosphoric acid
4. Nucleosidases – purine nucleosidase; pyrimidine nucleosidase
Nucleoside Nucleosidase sugar + base
Lesson 4. Bile
Bile
- Clear, golden yellow, slightly viscid fluid, secreted by the liver cells
- Average secretion of 500 – 700 ml/day
- Stored in the gall bladder where it undergoes concentration
- Gall bladder bile is more concentrated, more viscous and dark yellow-green in color
- Cholecystokinin – hormone which stimulates the discharge of bile into the bowel
(relaxation of the sphincter Oddi and Contraction of the gall bladder)
- Cholagogue – substance that stimulates the flow of bile
➢ Solid content (1 - 4%)
*Organic components (3/4) ▪ Mucin, Urea, ALP
▪ Bile acids *Inorganic
▪ Bile pigments ▪ HCO3-
▪ Bilirubin ▪ Cl
▪ Cholesterol ▪ Na
▪ Small amount of ▪ K
▪ phospholipid (Lecithin)

Liver Bile Gall Bladder Bile
▪ 1-4% ▪ Solids: 4 - 17%
▪ Less viscous ▪ more viscous
▪ Golden yellow ▪ dark yellow-green
▪ pH 7 - 8.5 ▪ pH 5.5
Bile acids
1. Cholic acid – 26 – 60% of the total
2. Chenodeoxycholic acid – 30 – 50%
3. Deoxycholic acid – 5 – 25%
o These bile acids are conjugated with glycine or taurine to form bile salts
Functions of bile
1. Activate steapsin
2. Aid in absorption of fat and fat-soluble substances by forming water-soluble complex
(hydrotropic effect)
3. Enhance digestion of fat (emulsification)
- lowers surface tension, and disperse in an emulsion which enhances digestion of fat
4. Choleretics (stimulate secretion of bile)
5. Maintain cholesterol in solution
Bile Pigments
▪ From the degradation of hemoglobin
➢ Biliverdin
➢ Bilirubin (formed from the reduction of biliverdin)
Activity 3. Given the following diagram, discuss your understanding of it by relating it to

previous discussion on bile pigments. Make your discussion concise but complete.
____________________________________
____________________________________
____________________________________
____________________________________
____________________________________
____________________________________
____________________________________
____________________________________
____________________________________
____________________________________
________________________
Absorption
- Occurs almost exclusively in the small intestine
- Simple sugars, amino acids, short chain fatty acids, and glycerol are absorbed into blood
stream via capillary network of villi. Products of lipid digestion are absorbed as chylomicrons
into intestinal lymphatics via central lacteal villi

Activity 3. Given the following food items, identify where the site of chemical digestion starts:
(mouth/stomach/intestines/none)
1. pancake _____________ 4. toasted bread _____________

2. bacon _____________ 5. siomai _____________
3. pan de sal _____________
Lesson 5. Chemical Changes in the Large Intestine
D. LARGE INTESTINES
While its true that the large intestines produce no digestive enzymes, it is still a part of
the digestive system. It functions primarily on the absorption of water and electrolytes and the
elimination of feces. Note however, that there are chemical changes that occur in the large
intestines.
Chemical Changes in the large Intestines

o The semi-liquid residue which has escaped digestion and absorption are passed into
the large intestines. The presence of intestinal microorganisms disintegrates these
organic residues into simpler fragments
1. Fermentation
- Bacterial degradation of carbohydrates under anaerobic conditions
- Products are
o Organic acids
▪ Lactic acid
▪ Formic acid
▪ Propionic acid
▪ Succinic acid
o Gases
▪ Methane
▪ Carbon dioxide
▪ Hydrogen gas
2. Putrefaction
- Bacterial decomposition of protein under anaerobic conditions
Tryptophan Indole + skatole
3. Deamination
- Removal of amine group from simple amino acids to form short-chain organic acids
- R - CH - COOH → R - CH2 - COOH
NH2
4. Decarboxylation
- Removal of carboxyl group forming amines
R - CH - COOH → R - CH - NH2
NH2

Detoxification
- Includes all processes by which noxious substances are rendered less harmful or more
easily excreted
Reactions Involved
1. Oxidation
- One of the most important means of detoxification
- Example:
Ethyl alcohol + Oxygen = carbon dioxide + water
2. Reduction – has minor role in detoxification
3. Hydrolysis – some drugs used for therapy are hydrolyzed in the body
Acetylsalicylic acid = salicylic acid + acetic acid
4. Conjugation
- Process called upon whenever oxidation becomes ineffective
- Brought about by the combination of the toxic substances or one of its metabolites with a
compound occurring normally in the kidney
- Occurs chiefly in the liver
- Conjugating agents
o Acetic acid
o Cysteine
o Glucoronic acid
o Glutamine
o Glycine
o Methyl group
o Sulfuric acid
o Thiosulfate
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Summary:
Digestion:
• Begins in the oral cavity
- Salivary amylase – starch
- (lingual lipase)
• CHON digestion begins in the stomach
- HCl and Pepsin
- Gastric lipase
• Digestion continues in the Intestine
• Intestines:
- BILE emulsifies, neutralizes and excretes cholesterol & bile pigments
- PANCREATIC SECRETION contains enzymes for attacking all the major foodstuffs
- INTESTINAL SECRETIONS complete the digestive process
- THE MAJOR PRODUCTS OF DIGESTION ARE ASSIMILATED
- BACTERIA in the LI cause putrefaction & fermentation
Assessment:
Given the following descriptions or definitions, identify what is asked. Write your answers
before the number. ERASURES and WRONGLY SPELLED WORDS will not be corrected.
_____________________________________ 1. What enzyme activates trypsinogen?

_____________________________________ 2. This enzyme is capable of curdling milk in
adults.
_____________________________________ 3. Comparing saliva, gastric and pancreatic
juice, which will have the highest percentage
of water present?
_____________________________________ 4. Which in number 3 has the highest
percentage of solids present?
_____________________________________ 5. What specific juice or substance has the
most alkaline pH among all body fluids?
_____________________________________ 6. What enzyme works against the carboxyl
group of the C-terminal?
_____________________________________ 7. Salivary glands that produce mucin.
_____________________________________ 8. The thought of food influences saliva
secretion, what is this factor called?
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_____________________________________ 9. What pathological variation of HCl is
brought about by low production of HCl?
_____________________________________ 10. Which cell in the gastric glands produces
pepsin?
_____________________________________ 11. TRUE or FALSE. Ptyalin is inactivated by
HCl.
_____________________________________ 12. What enzyme hydrolysis RNA?
_____________________________________ 13.What is the active form of
chymotrypsinogen?
_____________________________________ 14. Pancreatic duct joins common bile duct
and enters duodenum at _____.
_____________________________________ 15. TRUE or FALSE. The hydrolysis of starch
in the mouth produces maltose.
_____________________________________ 16.Which among bread, oil and egg will
undergo physical digestion in the mouth?
_____________________________________ 17. The bile pigments are derived from the
degradation of ____.
_____________________________________ 18. This is the main function of bile.
_____________________________________ 19. What is the product of decarboxylation?
_____________________________________ 20. TRUE or FALSE. Large intestines take
part in digestion.
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LABORATORY:
INTRODUCTION:
Saliva is a mixture of the secretions of the numerous buccal glands. It is a colorless slightly
opalescent fluid. The composition varies with the relative proportion of the secretions of the
different glands. The secretion of the parotid is more watery, rich in ptyalin, while those of the
submandibular and the sublingual are more viscid containing more of mucin, a glycoprotein.
Name: _____________________________ Date: _____________________________

ACTIVITY NO. 8
DIGESTION: SALIVA
OBJECTIVES:

1. identify correctly the inorganic and organic constituents of saliva
2. Research on the positive results of tests to detect the presence of organic and
inorganic constituents of saliva using qualitative procedures
3. Appreciate wholly the importance of saliva in digestion
MATERIALS:
Reference materials Pen/Pencil

Scoring rubric
For Procedures:
o 2 points for correct downloaded picture with label
- 1 point is given for items that has answers only
Collection of Specimen. The following is the supposed to be step in collecting saliva to test for
the presence of organic and inorganic matter. You are advised to read through it to be able to
appreciate the concept of the activity. You are not required though to collect the said specimen
since the activity will focus more on the expected result.
1. Collect 20 mL of saliva in a wide mouthed plastic vial and then cover until ready for use
Preparation of Mucin. The following is the supposed to be step in preparing the mucin which will
be used to test for the presence of organic matter in the saliva. Remember that in the lecture, it
was mentioned that there are different components of the digestive juices. The purpose of this
activity is to check whether or not, the different organic and inorganic matters are really present
in the saliva. While the activity cannot be performed in your house, it’s still best to be familiar with
the procedure.
➢ Get 10 mL of saliva from your sample and place this in a 50 mL beaker
➢ Add 30 mL of cold 95% ethanol (you may place the reagent bottle of ethanol in a trough
filled with ice). Stir
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➢ Cover with parafilm and let stand until a distinct whitish mucoid precipitate is formed
➢ You may fish out the precipitate using an applicator stick or a Pasteur pipette or you may
pour off the supernatant liquid
➢ Place the precipitate on a watch glass and divide it into 4 parts
1. Biuret test. Should a test tube with mucin is added with NaOH and CuSO 4, what would
be the possible result. Download a photo of the positive result and attach it to your paper.
2. Millon’s test. Should a test tube with mucin is added with Millon’s reagent and heated in
a water bath for several minutes, what would be the possible result. Download a photo of
the positive result and attach it to your paper.
3. Molisch’s test. Should another test tube with mucin id added with Molisch’s reagent and
concentrated sulfuric acid, what would be the possible result. Download a photo of the
positive result and attach it to your paper.
4. Benedict’s test. If a test tube with mucin is added with Benedict’s reagent and heated in
a boiling water bath for 3-5 minutes, what would be the possible result. Download a photo
of the positive result and attach it to your paper.
Test for the Inorganic matters. The following procedure is done to prepare the saliva for tests
of the presence of inorganic matters.
➢ Place 10 mL of the remaining saliva in a 50 mL beaker
➢ Acidify with nitric acid and heat to boiling. Filter and save the filtrate.
1. Test for Chloride. Should a test tube with filtrate is added with nitric acid and silver nitrate,
what would the possible result be? Write your answer in the table below and download a
photo of the positive result and attach it to your paper.
2. Test for Sulfate. Should a test tube with filtrate is added with conc. HCl and barium
chloride solution, what would the possible result be? Write your answer in the table below
and download a photo of the positive result and attach it to your paper.
3. Test for calcium ions. Should a test tube with filtrate is added with ammonium oxalate,
4. Test for Nitrite. Should a test tube with filtrate is added with conc. Sulfuric acid, KI solution
and starch solution, what would the possible result be? Write your answer in the table
below and download a photo of the positive result and attach it to your paper.
5. Test for Thiocyanates. Should a test tube with filtrate is added with ferric chloride and
conc. HCl, what would the possible result be? Write your answer in the table below and
download a photo of the positive result and attach it to your paper.
6. Test for Phosphates. Should a test tube with filtrate is added with ammonium molybdate,
OBSERVATIONS
Ion or Name of test or Reagent Result/ Conclusion
component used interpretation
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Scoring Rubric
1. What is the importance of saliva in digestion?
2. Name the glands that produce saliva and describe the characteristics of the saliva they
secrete
3. Give the importance of each of the following constituents of saliva:
a. Chloride Ion: Cl – 1
b. Thiocyanate Ion? SCN – 1
c. Mucin
d. Ptyalin or Amylase
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LABORATORY:
INTRODUCTION:
Bile is the yellowish-brown fluid secreted by the liver and discharged into the duodenum
where it aids in the emulsification of fats. It is a viscid fluid with an alkaline reaction. Its important
constituents are bile acids, bile pigments, inorganic salts and cholesterol.
The first step in fat digestion is to break the fat globules into small sizes so that the water –
soluble digestive enzymes can act on the globule surfaces. This process is called emulsification
of the fat and it is done under the influence of bile acids or salts. The carboxyl group of the bile
salt is water soluble, whereas the sterol portion is highly soluble in fat. Bile salts thus allow the
formation of colloidal dispersions between fat and water – soluble enzymes. This greatly
decreases the interfacial tension of the fat breaking them into minute particles thereby
increasing the total surface area of the fat particle to be acted upon by digestive enzymes.
Name: _____________________________ Date: _____________________________

ACTIVITY NO. 9
DIGESTION: BILE
OBJECTIVES:
1. describe correctly the physical and chemical properties of bile
2. research on the specific tests for the components of bile
3. trace properly how bile pigments are formed
MATERIALS
Refence material Pen/pencil
Scoring rubric
For Procedures:
o 2 points for correct downloaded picture with label
- 1 point is given for items that has answers only
Physical properties
1. Research on the following physical properties of bile:
a. Color: __________________
b. Odor: __________________
c. pH: ____________________
Test for the organic constituents. The following are tests used to determine the presence of
organic materials in bile. Read the instruction for each part and do what is asked.
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Test for bile acids:
1. Should a test tube with bile is diluted with 0.5 mL of water, added with 5% sucrose
solution and eventually conc. Sulfuric acid, what is the possible result. Write it in the
observation. Download a photo of the positive result and attached it to your paper.
Observation
___________________________________________________________________
What is the name of this test?
___________________________________________________________________
What is the principle behind this test?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Test for Bile Pigments

1. Should a test tube with bile is diluted with 0.5 mL of water, placed in a filter paper in the
middle of a watch glass and then added with conc. Nitric acid, what is the possible
result? Download a photo of the possible result and attach it to your paper.
What are the colors produced?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Name the bile pigments responsible for the colors produced:

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Test for bilirubin

1. Should a test tube with bile is diluted with 1 mL of water and added with barium chloride
solution, what is the possible result. Write your answer in the observation.
Observation:
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
What is the rationalization behind this result?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
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2. Should you filter the product obtained in procedure 1 and add Fouchet’s reagent to the
filter paper with precipitate, what is the possible result? Write it in the observation below
and download a photo of the positive result and attached it to your paper.
Observation:
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Test for Cholesterol
Procedure A
1. Should a test tube with bile is dissolved with chloroform and added with conc. Sulfuric
acid, what is the possible result. Write it in the observation below and download a photo
of the positive result and attach it to your paper.
Observations:
___________________________________________________________________
___________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Procedure B
1. Should a test tube with bile is dissolved with chloroform and then added with acetic
anhydride and eventually sulfuric acid, what is the possible result. Write it in the
observation below. Download a photo of the positive result and attach it to your paper.
Observations:
___________________________________________________________________

___________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Test for Inorganic constituents. The following is the step to make a fusion mixture to test the
presence of inorganic material in bile. Read through it to understand the concept.
1. In an evaporating dish, mix 2 g of Na2CO3 and 1 g of KNO3, this is your fusion mixture
2. Add 15 mL of bile to the fusion mixture
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3. Heat strongly until the mixture is charred or burnt black.
➢ If the laboratory is equipped with a hood, you might consider doing this under it
since the odor could become obnoxious, otherwise you can use masks while
heating
4. Cool it enough to be handled and add 10 mL of water
5. Filter and divide the filtrate into 3 equal parts and place separately in test tubes
6. Test for chlorides, phosphate ions and sulfate ions (refer to the experiment in saliva for
the procedures.) Write the possible result in the table below and download photo for
each procedure and attach it to your paper.
OBSERVATIONS
IONS TEST REAGENTS RESULTS

-
Cl
PO4 -3
SO4 -2
Aside from the ions tested, name 3 other ions found in bile.
____________________________________________________________________________
____________________________________________________________________________
Scoring Rubric
1. How is bilirubin metabolized by the body? Use a flowchart to start your discussion.
2. What is the most important constituent of bile from the physiological point of view? Give
at least four (4) important functions of it.
3. Name the three (3) most abundant Bile acids in human. What is their importance in
digestion?
4. What is the danger of excessive amount of cholesterol in the bile?
5. What possible disease/s are associated with bile pigment abnormalities? Discuss one
only.
Digestion
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WEEK NO. 7
SUBMODULE NO. 7
INTRODUCTION TO
CARBOHYDRATES
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LESSON: Unit 7: Carbohydrates
TOPICS:
6.1. Definition and General structure of CHO
6.2. Importance and Functions of Carbohydrates
6.3. Classification of Carbohydrates
6.3.1. Monosaccharides
6.3.2. Disaccharides
6.3.3. Polysaccharides
LECTURE: 5 hours
LABORATORY: 4 hours
1. define correctly the terms associated with carbohydrates
2. Describe completely CHO according to composition, classification and chemical
properties
3. Demonstrate accurately laboratory safety practices and proper waste disposal;
4. Precisely perform laboratory activities
5. Explain accurately the chemical reactions, which underlie the results, obtained in the
biochemistry laboratory and relate its importance in the clinical aspect of the course.
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LESSON PROPER
INTRODUCTION:
The name carbohydrate originally meant a compound with the empirical formula CH2O,
literally, a hydrate of carbon. This definition has been broadened to include aldehydes and
ketones having two or more hydroxyl groups. Among the compounds that fall into the class of
carbohydrates are the starches, cellulose, glycogen and the sugars. The carbohydrates are
important source of energy for all organisms and form the supporting tissue of plants and some
animals. There are three important classes of carbohydrates: the monosaccharides,
disaccharides and the polysaccharides. (Holtzclaw and Robinson, 1988)
CARBOHYDRATES
- Commonly known as sugars

- They are defined as polyhydroxy aldehydes or polyhydroxy ketones or substances that
yield these compounds when hydrolyzed
IMPORTANCE AND FUNCTIONS OF CARBOHYDRATES
1. Energy Yielding Nutrients – they serve as primary source of metabolic fuel needed by
a living body and as means to store and reserve energy in the form of glycogen
2. Serve as structural components of the living cells - as carbohydrates combine with
proteins, they from glycoproteins and proteoglycans that are very important components
of the cell membrane
3. Serve as structural framework – for DNA and RNA as part of nucleotides
4. Mediate Interaction between cells – blood types are determined by specific membrane
bound CHO
5. Building materials – cellulose is found in wood, cotton, paper
CLASSIFICATION OF CARBOHYDRATES
- The simplest structural unit of carbohydrate is known as saccharide

- In Latin, Saccharine means sugar
A. According to number of saccharide units

- Monosaccharides
o The simplest carbohydrate – has only one saccharide unit
o Cannot be broken into smaller CHO molecules by hydrolysis
- Disaccharides
o Contains two monosaccharide units
o It can be hydrolyzed to give two monosaccharide units
- Oligosaccharide
o Polymers made up of two to 10 (2-10) monosaccharide units
- Polysaccharides
o Giant polymers made of many monosaccharide units
o The most complex CHO because they contain more than 10 saccharide units
B. According to number of carbon atoms
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- Trioses – with three carbon atoms
- Tetroses – with four carbon atoms
- Pentoses – with five carbon atoms
- Hexoses – with six carbon atoms
- Heptoses – with seven carbon atoms
Theoretically, a monosaccharide can have any number of carbons

greater than three, but only monosaccharide of three to seven carbons
are commonly found in the biosphere
C. According to its functional group
o The functional group of carbohydrates

is the carbonyl group (-C=O)
- Aldose - polyhydroxy aldehyde and the
carbonyl group is found at one of the terminal
sides of the hydrocarbon chain
- Ketose - polyhydroxy ketone and the
carbonyl group is found in between 2
hydrocarbon groups
https://www.mikeblaber.org/oldwine/BCH4053/Lecture12/Lecture12.htm
http://www.hammiverse.com/lectures/5/1.html
D. As a furanose or pyranose
- Depends on whether the cyclic structure of the carbohydrate is related to that of the five
or six-membered ring compound furan or pyran respectively
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https://www.quora.com/What-are-the-differences-between-furanose-and-pyranose
E. As Alpha or beta configuration

- Based on the –H and –OH groups about a specific chiral carbon in the cyclic form of the
monosaccharide
Epimers:
- two sugars which differ only in the configuration around a single C atom
https://socratic.org/questions/can-anyone-explain-to-me-how-to-identify-epimers-and-anomers-in-carbohydrates-su
Asymmetric Carbon
- refers to the C atom in the structure of a sugar to which 4 different radicals are attached.
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https://www.quora.com/What-is-the-difference-between-symmetric-asymmetric-carbon-atom-in-carbohydrates
• the asymmetric carbon gives rise to the following:
Optical activity/ Mutarotation
- the ability of a sugar solution to bend or deflect plane of polarized light.

- (d or + ) DEXTROROTATORY (to the right)
- ( l or - ) LEVOROTATORY (to the left)
Enantiomers (mirror-image isomers)

- Stereoisomerism/ enantiomerism
o Depends on the spatial
orientation of the –H and
–OH group attached to
the carbon atom adjacent
to the terminal primary
alcohol group
o the ability to form two
sugars which are mirror
images to each other (D-
isomer or L-isomer).
o The configuration is
based on the location of
the –OH linked to the C
atom which is the next to
the terminal.
https://slideplayer.com/slide/12282699/
• D-isomer: -OH is on the RIGHT of the C atom adjacent to the terminal
primary alcohol group
• L-isomer: -OH is on the LEFT of the C atom adjacent to the terminal
primary alcohol group.
MONOSACCHARIDES
- Simplest carbohydrates which cannot be broken down into smaller carbohydrate
molecules
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1. Trioses (C3H6O3) – glyceraldehyde, dihydroxyacetone
2. Tetroses (C4H8O4) – erthyrose, erythrulose
3. Pentoses (C5H10O5) – xylose, ribose, deoxyribose, arabinose, rhamnose
4. Hexoses (C6H12O6) – glucose, galactose, mannose, fructose
5. Heptoses (C7H14O7) – mannoheptose, mannoheptulose
Chemical structure of monosaccharides
1. The carbon skeleton is unbranched

2. All the Carbon atom except one has a hydroxyl (-OH) group
3. One has a carbonyl oxygen which may be in terminal position to give rise to an aldehyde
(aldose), or may be located on the second carbon atom giving rise to an ketone (Ketose)
4. The terms aldose for hydroxyaldehydes and ketose for hydroxyketones specify the
functional group of a monosaccharide
5. The general formula of carbohydrates: Cn(H2O)n
Where n = number of carbon atoms
6. It includes both the aldehyde and the ketone groups (functional groups)
7. Hydroxyl groups are attached to each C atoms, except that of the carbonyl group
IMPORTANT MONOSACCHARIDES
1. Diose – a monosaccharide with 2 carbon sugars

- They are the simplest carbohydrates
o Example: Glycoaldehyde
2. Triose – a monosaccharide with three carbon atoms
- Glyceraldehyde
o Produced from the breakdown of larger monosaccharides
o D-glyceraldehyde with D-glyceraldehyde – 3 – phosphate is an intermediate in
the metabolic pathway of glycolysis (conversion of glucose to lactic acid)
3. Tetroses – carbohydrates containing 4 carbon atoms
- Examples
o Erythrose – aldehyde tetrose/aldotetrose
o Erythrulose – ketone tetrose/ketotetrose
o Threose – aldehyde tetrose/aldotetrose
4. Pentose – monosaccharides with 5 carbon atoms, the most widely distributed sugars in
the living systems
- Ribose
o Component of ribonucleic acid (RNA), RNA sugar
o RNA – is a polymeric compound that plays a major role in the synthesis of
protein molecules
- Deoxyribose
o DNA sugar
o DNA – is a compound which transmits genetic information from parent to
offspring by directing the synthesis of protein molecules
- Xylose
o Found in nuts
o Also known as sugar alcohol
o It is a sugar that is not normally metabolized
o It is used to detect CHO malabsorption
• Xylose excretion test
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• After a known amount of D-xylose has been given to a patient, its
appearance in the urine is observed
• In normal individuals, 65% of the xylose should be absorbed intact
and excreted in the urine in 5 hours
• A decreased amount of D-xylose in the urine indicates
malabsorption due to intestinal disease
- Arabinose
o Found in nuts
o Known as gum sugar
5. Hexoses – monosaccharide with 6 carbon atoms

o The aldohexoses, ketohexoses and aldopentoses will receive special attention
since they include the most important monosaccharides in living systems
- Glucose (grape sugar/blood sugar/plasma sugar/dextrose)
o Most common source of cellular ATP and from which most complex
carbohydrates are made from
o Natural sugar mostly in the D form
Blood concn: 80 – 100 mg/dL
Glucose - exists in D-glucose: aka Dextrose because it is

dextrorotatory;
- does NOT require further digestion and may be
given IV to patients who cannot take in food by mouth
- D-Galactose
o brain sugar – D-glucose is found in the brain and nervous tissue in the form of
glycolipids
o An aldohexose; Less than half as sweet as glucose
o Not a natural sugar
o Has the fastest rate of absorption in the intestines
- D-Fructose (levulose/fruit sugar)

o Sweetest sugar
o A natural ketohexose; occurs in honey
o Serves as a source of energy for the sperm cells
o Liver can convert fructose to glucose
Fructose: Present in honey in equal amount with glucose. It is

sometimes used as dietary sugar because less is needed with
the same sweetness
- Levulose is another name for it because it is
levorotatory
- Mannose
o D-Mannose is the 2-epimer of glucose
• exists primarily as sweet-tasting α- (67%) or as a bitter-tasting β- (33%)
anomer of the pyranose
o Naturally-occuring aldohexose
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o Not found free in nature but is widely distributed in the form of polysaccharide.
o L-Mannose is not normally used in biological systems
DISACCHARIDES
- Composed of two saccharide groups
- Formed when two monosaccharide molecules combine by splitting out a molecule of
water
- The glycosidic linkage that joins the two sugars always involves the hemiacetal –OH
group of one sugar and one of the OH groups of the other monosaccharide
https://ib.bioninja.com.au/standard-level/topic-2-molecular-biology/23-carbohydrates-and-lipids/sugar-subunits.html
Important Disaccharides
1. Maltose
- Malt sugar
- Broken down into 2 molecules of glucose
- Linkage: alpha - 1,4
- Found in combined form as starch and is one of the intermediate products in the
digestion of starch
- Derived from “malting” (soaking, germinating and drying of grains such as barley)
- The malt produced includes malt starch and enzymes that are able to convert malt
starch into maltose, which is then fermented to produce alcohol
https://chem.libretexts.org/Courses/Sacramento_City_College/SCC%3A_Chem_309_-
_General_Organic_and_Biochemistry_(Bennett)/Text/14%3A_Carbohydrates/14.6%3A_Disaccharides
2. Cellobiose
- Composed of 2 glucose molecules
- Linkage: beta – 1,4
- Produced from the breakdown of the polysaccharide cellulose
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o Mammalian digestive tracts do not secrete enzymes that can break the beta –
1,4 linkages between 2 glucose molecules
https://commons.wikimedia.org/wiki/File:Formation_du_cellobiose.PNG
3. Lactose
- Also known as milk sugar
- Made by combining B-D-Galactose and D-glucose
- Linkage: Beta-1,4
- In the body, it is broken down by the action of the enzyme known as LACTASE
- Deficiency of the enzyme leads to inability to absorb lactose and the condition called
LACTASE DEFFICIENCY SYNDROME or LACTOSE INTOLERANCE
- This condition is controlled by eliminating all dietary sources of lactose (e.g. dairy
products, some medications)
https://commons.wikimedia.org/wiki/File:Formation_du_lactose.PNG
4. Sucrose
- Also known as table sugar
- Made by combining alpha-D-glucose and beta-D-fructose
- Linkage: alpha,beta - linkage
- Considered as a non-reducing sugar, a sugar which does not react with Tollen’s or
Benedict’s reagent
- Main sources: sugar and sugar beets
- When sucrose undergoes hydrolysis, it produces an equimolar mixture of glucose and
fructose called INVERT SUGAR through the action of the enzyme SUCRASE or
INVERTASE
https://bio.libretexts.org/Courses/Lumen_Learning/Book%3A_Biology_for_Non-
Majors_I_(Lumen)/03%3A_Important_Biological_Macromolecules/3.03%3A_Carbohydrates
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POLYSACCHARIDES
- General formula: (C6H10O5)x
- Made up of several saccharide groups
- they do not exhibit reducing properties
- Made up of several hexose molecules minus the corresponding molecules of water
Properties of polysaccharides
- White, tasteless, amorphous compunds
- X-ray analysis: crystalline
- No reducing property; do not form osazone crystals
- High MW; mostly are insoluble
- Non-fermentable by yeast
- Upon hydrolysis, yield simple sugars and sugar derivatives
- Polysaccharides with large MW are antigenic
Homopolysaccharides
- These are polysaccharides which on hydrolysis yield only one kind of monosaccharide
1. Starch – found abundantly in the plant kingdom particularly in fruits, cereals, seeds,
bulbs and tubers
- Most important constituent of the human diet
- Occurs in the form of granules
https://www.slideshare.net/thelawofscience/carbohydrates-intro-11699082
2. Inulin
- Found in the bulb of onions and garlic
- White, odorless, tasteless powder soluble in hot water
- Upon hydrolysis (by the action of either acid or the enzyme INULASE) yields fructose
- Clinical significance: study of renal function; inulin is easily excreted through the
kidneys when injected intravenously
3. Glycogen
- Also called animal starch

- Found in the liver as storage material and in the muscles as source of energy
- Occurs in yeast, algae and fungi; abundant oysters
- Structure is similar to that of amylopectin except that branching (via alpha-1,6 linkages)
occurs about 8-12 glucose units (24 – 30 glucose units for amylopectin)
• Glycogen amylase maltose

• Glycogen acid glucose
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- Monosaccharides like glucose are synthesized and stored in the muscles and liver as
glycogen
- When the body needs them for heat and energy purposes, they are broken down to D-
glucose to supply the physiological requirement
4. Dextrin
- Intermediary products of starch hydrolysis
- Amorphous, white powder
• Dextrin amylase maltose
• Dextrin acid glucose
- Uses:
o Used as mucilage
o Used in infant feeding; prevents the formation of large, heavy curds or milk in the
baby’s stomach, thus facilitating digestion
o Found in some breakfast foods and malt preparation
5. Cellulose
- Most abundant organic compound
- Forms the supporting structure of plants
- Purest source: cotton
- Negative to the iodine test
- Structure: the monosaccharide units of cellulose are beta-glucose molecules connected
by beta-1,4 linkages into cellobiose units
o Humans cannot digest cellulose due to the absence of the enzyme capable of
cleaving the beta-1,4 linkage
https://www.slideshare.net/thelawofscience/carbohydrates-intro-11699082
- Uses of Cellulose
o Rayon – fiber made from cellulose
o Cellophane – transparent cellulose sheeting
o Cellulose + HNO3 + H2SO4 = nitrocellulose
(cellulose nitrate)
• With 12.5 – 13.4% N = guncotton
• With 11 – 12% N = celluloid (used in plastics and photographic films)
o Cellulose + acetic acid (or acetic anhydride) = cellulose acetate
• Used as a substitute for celluloid
• Fabric, upholstery and cigarette filters
o Cellulose plays an important role in human physiology by furnishing bulk or
roughage which stimulates peristalsis, thus promoting the evacuation of the
bowel
6. Hemicellulose
o Hydrolyzed upon boiling with mineral acids
o Products: pentoses and hexoses
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a. Pentosans (C5H8O4)x: widely distributed in the plant kingdom
• E.g gum Arabic
• Liberates pentose upon hydrolysis
b. Hexosans (C6H10O5)x: represented by galactans (widely distributed in plants)
and give rise to galactose upon hydrolysis
• E.g. agar-agar has very little food value but has been found to be very
efficient in relieving chronic constipation; also used in making culture
media
c. Hexo-pentosans: e.g. pectin – colloidal CHO responsible for the jellying
properties of fruits
Heteropolysaccharides
- Yield mixtures of monosaccharides and derived products
- Nitrogen-containing mucopolysaccharides
o Neutral mucopolysaccharide
• Made up of N-acetyl-hexosamine and hexose
• E.g. those occurring in bacteria and the so-called “mucoids” including
important immunological specific blood group substances
o Acid mucopolysaccharide
• Contains acetylhexosamine, hexuronic acid, sulfate or phosphate
• examples of mucopolysaccharide:
• Those containing sulfate
o Chondroitin sulfate of the cartilage tissue - contains N-
acetylgalactosamine, glucuronic acid and sulfate
o Chondroitin sulfate of the skin – has 1-iduronic acid
instead of glucuronic acid
o Heparin – glucosamine N-sulfate, glucuronic acid and
sulfate
o Hyaluronic acid
• Main constituent of the ground substance of connective tissues
• Found in synovial fluid, pleural fluid, vitreous humor and Wharton’s jelly
• Made of d-glucuronic acid, d-glucosamine and acetic acid
• Function:
• Lubricant
• Cementing substance which allows passage of metabolites but
not of the infective organisms
• It is fragmented by hyaluronidase (spreading factor), an enzyme found in
bacteria, sperm and in the poisonous secretions of reptiles and other
animals
o Heparin
• Generated by certain types of cells lining arterial blood vessels and by the
lung tissues
• Powerful inhibitor of blood clotting thus preventing intravascular
coagulation
• In practice, it is used to prevent clotting of blood specimens
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FORMATIVE ASSESSMENT: None
ASSIGNMENT:
Activity:
Materials: puncher/scissors, colored papers, bond papers, glue
- Using your puncher/scissors, punch/cut out holes in your colored papers. The circles will
represent glucose units.
- Using the circles, create the structure of the following showing the branching accurately
o Amylose
o Amylopectin
o Glycogen
Carbohydrates are also known as_______________________.

Carbohydrates are an abundant biomolecule.
• More than 50% of the carbon in organic compounds is found in carbohydrates
• Plants use photosynthesis to store energy in__________________, a simple sugar
6 CO2 + 6 H2O + Energy à C6H12O6 + 6 O2
Carbohydrates are a large class of naturally occurring polyhydroxy ___________________and
________________________.
• ________________________(also known as simple sugars) are the simplest carbohydrates

containing 3-7 carbon atoms. A sugar containing:
– an aldehyde is known as an ______________________
– a ketone is known as a _________________________
Classification of Carbohydrates
Carbohydrates are grouped into 3 classes:

• ___________________________ are the simplest sugars and serve as the building blocks of
larger molecules – Example: Glucose
• __________________________ contain 2-10 monosaccharides bonded together
(building block = residue) – Example: Sucrose
• _____________________________contain more than 10 residues – Example: Complex
Carbohydrates
Naming Monosaccharides
Carbohydrate nomenclature is unique to “sugar chemistry” — we do not name

monosaccharides using the IUPAC rules.
• Monosaccharide names end in “ose."
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• Monosaccharides can be classified by:
a. Carbonyl group: ______________________ or _______________________
b. ______________________ of carbons: triose, tetrose, etc.
• Both: aldotriose, ketotriose, and so on…
Stereoisomerism
Carbohydrates are ________________________molecules since they have carbon atoms

carrying four different groups.
• Enantiomers have the same _______________ except they behave differently in the way they
rotate polarized light and the way they are affected by catalysts.
D form and L form sugars.
• D: the –OH group on the chiral C furthest from the C=O comes out of the plane of paper
and points to the_______________________.
• L: the –OH group on the chiral C furthest from the C=O comes out of the plane of paper
and points to the ______________________.
Monosaccharides:
D-glucose, also called ___________________or_________________, ___________________
is the most important monosaccharide in human metabolism.
Disaccharides:
The glycosidic bond in maltose is referred to as an __________________ bond since the
monosaccharide on the left reacts it’s α-anomer hemiacetal at C-1 with a hydroxyl at C-4 on the
second monosaccharide.
Polysaccharides:
___________________________ contain 10 or more residues
o In a_________________________, all the residues are the same monosaccharide
o In a_________________________, the residues are built from more than one type of
monosaccharide
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Starch is a homopolysaccharide used by some plants to store energy; there are 2 components
of starch:
• 1) _____________________________
• 2) _____________________________
• In amylopectin (right), branches occur every __________ residues

• In glycogen (left), branches occur every ___________ residues
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LABORATORY
INTRODUCTION:
Carbohydrates are the most abundant class of organic compounds found in living organisms.
They originate as products of photosynthesis. Its primary function is to provide the body with
energy most especially our brain and nervous tissue.
Carbohydrates (also called saccharides) are molecular compounds made from just three
elements: carbon, hydrogen and oxygen. Monosaccharides (e.g. glucose) and disaccharides
(e.g. sucrose) are relatively small molecules. They are often called sugars. Other carbohydrate
molecules are very large (polysaccharides such as starch and cellulose.
Name: _____________________________ Date: _____________________________

Activity No. 10
CARBOHDRATES: PHYSICAL PROPERTIES AND GENERAL TESTS
OBJECTIVES
1. Differentiate accurately the physical appearance of carbohydrates

2. Discern correctly the solubility properties of carbohydrates using ordinary
solvents
3. Perform accurately the general tests for carbohydrates
4. Understand completely the principles behind these tests
MATERIALS: reference materials, tablespoons, teaspoons, clear containers
CHEMICALS: table sugar, cotton thread, cornstarch or gawgaw, water
Scoring rubric
For Procedures:
not clear
is not clear
o 0 – no hypothesis generalization or conclusion was made
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The following online reading materials may help:
1.https://chem.libretexts.org/Bookshelves/Introductory_Chemistry/Book%3A_The_Basics_of_G
OB_Chemistry_(Ball_et_al.)/16%3A_Carbohydrates/16.5%3A_Properties_of_Monosaccharides
2. https://www.angelo.edu/faculty/kboudrea/index_2353/Chapter_07_2SPP.pdf
3. http://dept.harpercollege.edu/chemistry/chm/100/dgodambe/thedisk/carbo/yback9.htm
4. https://byjus.com/chemistry/tests-of-carbohydrates/
Physical properties of Carbohydrates
Physical appearance and solubility

1. Using reference materials, note the physical appearance and the solubility of the
following carbohydrates in polar and non-polar solvents: Glucose, Fructose, ribose,
sucrose, lactose, starch, glycogen, cellulose
2. Write your answers on the table provided below.
3. You might also want to try dissolving a pinch of sugar in 2 tablespoons of water as well
as a pinch of starch (gawgaw) in 2 tablespoons of water to help you with some of the
answers. You can also use cotton threads for cellulose.
Observations:
Physical Appearance
Glucose
Fructose
Ribose
Sucrose
Lactose
Starch
Glycogen
Cellulose (cotton)
Solubility
Carbohydrates Solvents
Polar/water Non-polar
Glucose
Fructose
Ribose
Sucrose
Lactose
Starch
Glycogen
Cellulose
➢ Make a generalization about the solubility of carbohydrates in water/polar solvents

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
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➢ Make a generalization about the solubility of carbohydrates in non-polar solvents.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
General Tests for Carbohydrates
Molisch test
1. Place 0.5 mL of 5% glucose, fructose, ribose, sucrose, lactose, starch, cellulose, and
glycogen solution in appropriately labelled test tubes.
2. Add 2 drops of Molisch reagent to each of the test tubes and mix thoroughly
3. Incline the tube with glucose and let 0.5 mL of conc. sulfuric acid slide through the side
of the test tube so that two layers are formed, the lower layer being sulfuric acid
➢ Caution: sulfuric acid is corrosive
4. Repeat the preceding procedure with the rest of the test tubes
5. Using reference materials, what would be the color that will be produced at the junction
of the two liquids
6. Download the positive result for Molisch test and attach below this procedure
Anthrone test
1. Place 0.5 mL of 5% glucose, fructose, ribose, sucrose, lactose, starch, cellulose, and
glycogen solution in appropriately labelled test tubes.
2. Place 1 mL of Anthrone reagent into each of the test tube and mix
3. Let it stand until a color change is observed in the test tubes
4. Using reference materials, what would be the color that will be produced at the junction
of
5. Write your result on the data sheet provided for you
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6. Download the positive result of Anthrone test and attach below this procedure
Iodine test
1. Place 0.5 mL of 5% starch, cellulose, and glycogen solution in the appropriately labelled
test tube.
2. Add 1 – 2 drops of Lugol’s reagent into each of the test tube
3. Observe for any color changes
4. Using reference materials, what would be the color changes for the following
carbohydrates with iodine solution?
5. Record your result on the data sheet provided for you.
6. Download the result for iodine test on starch, cellulose and glycogen and attach it below
this procedure.
Observations
General tests for carbohydrates

Carbohydrate Molisch test Anthrone test Iodine test
Glucose
Fructose
Ribose
Sucrose
Lactose
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Starch
Glycogen
Cellulose (cotton)
- The sugar and starch solutions may be thrown safely down the drain, however the cotton
threads should be thrown in the garbage as it may clog your drain.
What conclusion can you derive from this activity?

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
________________________________________________
Scoring Rubric
1. What is the principle behind Molisch and Anthrone test?

2. How will you differentiate starch from glycogen structurally?
3. Write the structural formula of the monosaccharides in this experiment using Fischer
projection while for the disaccharides, use Haworth formula
4. How are glycosidic linkages formed?
5. Differentiate the glycosidic linkages
a. between glucose and fructose in sucrose
b. of glucose units in amylose, amylopectin, glycogen, starch.
c. Between starch and cellulose
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WEEK NO. 8
SUBMODULE NO. 8
PROPERTIES OF
CARBOHYDRATES
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LESSON: Properties of Carbohydrates
TOPICS:
6.4. General Properties of Carbohydrates
6.4.1. Physical properties
6.4.2. Chemical properties
6.5. Digestion and Absorption of Carbohydrates
TIME/SCHEDULE: Week no. 9
LECTURE: 4 hours
LABORATORY: 5 hours

1. Differentiate completely the properties of carbohydrates
2. Write correctly simple equations related to the chemical properties of carbohydrates
3. Discern properly the importance of the knowledge of metabolism in keeping oneself
healthy;
4. Appreciate thoroughly the significance of the metabolism of different biomolecules in the
maintenance of normal life processes;
5. Precisely perform laboratory activities
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LESSON PROPER
LABORATORY
INTRODUCTION:
Carbohydrates are polyhydroxy aldehydes or polyhydroxy ketones or any other

substances that yields these substances upon hydrolysis. They are formed in plants by the
process of photosynthesis.
Different carbohydrates exist in nature and they do not often respond in similar manner
to some reagents because of slight differences in their molecular structures. This difference in
reactions makes it possible to identify the specific sugar in a given sample.
The following activity will help you detect the presence of carbohydrates and determine
some of their properties
Name: ____________________________ Date: _____________________________

Date Assigned: ____________________ Date submitted: _____________________
Activity No. 11
QUALITATIVE SPECIFIC TESTS FOR CARBOHYDRATES
OBJECTIVES
At the end of the laboratory period, you should be able to
1. Distinguish correctly the chemical properties of carbohydrates

2. identify correctly the simple tests for their identification
3. Differentiate properly the sugars using the tests performed
Scoring rubric
For Procedures:
For Rationalization
not clear
not clear
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The procedures in this activity need not be performed. Use your reference
materials to help you with your answers.
Seliwanoff’s test:
*Watch: https://www.youtube.com/watch?v=88hWF1-RrTQ
1. Place 10 drops of glucose fructose, sucrose and lactose in appropriately labelled test
tubes
2. Add 1 mL of Seliwanoff’s reagent to the tubes and mix
3. Immediately place them in a boiling water bath
4. Remove from the water bath and place them in a beaker containing cold water. This is to
stop further reaction from the residual heat.
5. Using your reference materials and a guide from the video, what are the possible results
for the following sugars? Your observations should be the color produced while the
interpretation is either positive or negative.
6. Download a photo of the positive result for Seliwanoff’s test and attach it below this
procedure.
Observations
Carbohydrates Observations Interpretation

Glucose
Fructose
Sucrose
Lactose
Based on your observations above answer the following questions:
➢ What is the function of resorcinol?

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
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➢ What is the aim of using a strong acid?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
1. Which carbohydrate gave a positive result to Seliwanoff’s test (or the first one to react)?
___________________________________________________________________
2. To what classification of carbohydrates based on functional group does your answer
above belong to?
___________________________________________________________________
3. What generalization regarding Seliwanoff’s test can you derive from this?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
4. What is the result of testing sucrose with Seliwanoff’s reagent? Explain your answers by
giving reasons and possibly structures.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Mucic acid test
1. What is Mucic acid test? What group of sugars will give positive result to this test?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
____________________________________________________________
2. The procedure of mucic acid test involve heating the appropriate sugar with
concentrated nitric acid for an hour and leaving the solution to stand overnight or so.
Then the following laboratory period, the test tubes will be noted for the formation sandy
crystals.
3. Suppose 3 mL of glucose, galactose, lactose and sucrose is added with 1 mL of conc.
HNO3, and heated for an hour, and the solutions stood overnight. What would be the
expected results? Use your reference materials as guide
Observations:
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
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5. What generalization can you derive from your observations?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
6. What is the result of testing lactose with mucic acid formation? Explain your answers by
giving reasons and possible structures.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
7. What simple test can you use to determine if the crystals formed is truly mucic acid?
______________________________________________________________________
______________________________________________________________________
Download the positive result for mucic acid test and attach it below this procedure.
Bial’s test:
Watch: https://www.youtube.com/watch?v=JboA8Ghyz5A
1. Use your reference materials and a guide from the video to derive your answers for the
following procedure.
2. Get 4 test tubes and label them as follows: Glucose, Fructose, Arabinose and ribose
3. Add 2 mL of Bial’s reagent to all test tubes
4. Add 5 drops of the corresponding sugar
5. Heat in a water bath for 5 – 10 minutes
6. Note the color that is produced.
Observations:
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
Name the sugars that gave a blue to green color

___________________________________________________________________
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What is the classification of the sugars that gave positive result to Bial’s test based on
the number of carbon atoms?
___________________________________________________________________
What generalization can you derive from your observations?
______________________________________________________________________
______________________________________________________________________
_____________________________________________________________
Is it possible to distinguish DNA and RNA structures using Bial’s test? Explain your
answer.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
____________________________________
Download the positive result of Bial’s test and attach it below this procedure
Scoring Rubric
1. Differentiate a monosaccharide from a disaccharide?

2. State the principle of the tests used in this activity.
3. How is Bial’s reagent prepared?
4. Glucose and galactose are epimers. What test can be used to differentiate these two
sugars? Explain your answer
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GENERAL PROPERTIES OF CARBOHYDRATES
Physical properties
1. Appearance
- Mono and disaccharides – white and crystalline
- Starches – amorphous powder
- Cellulose - fibrous
2. Solubility (in ordinary solvents)
- Inversely proportional to the complexity of their structures
o Mono and di – more soluble in water
o Higher CHO (e.g. starch) – insoluble, form colloidal solutions
o Cellulose – practically insoluble
3. Relative Sweetness of Sugars
sugars Relative Sugar substitutes (common Relative
sweetness brand names) sweetness
4
Fructose 100 Sucralose (Splenda®) 3.5 x 10
4
Invert sugar 75 Saccharin (sweet n low®)
1.7 x 10
4
Sucrose 58 Acesulfame potassium
1.2 x 10
(Sunette®, Sweet One®)
4
Glucose 43 Aspartame (Equal®,
1.2 x 10
Nutrasweet®)
Maltose 19
Galactose 19
Lactose 9.2
Chemical Properties
1. Reducing power
- All mono and di containing the potentially free aldehyde or ketone group possesses
reducing properties; reduce alkaline metals and are transformed into organic acids
Tests Reagents Positive result
1. Benedict’s test CuSO in NaOH Reddish brown to red

4
Na-citrate precipitate
CuSO + NaOH Cu(OH) + Na SO

4 2 2 4
Cu(OH) + reducing sugar + heat Cu O + H O + O
2 2 2
(reddish brown to red ppt)
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2. Fehling’s test CuSO4 in NaOH Reddish brown to red
Na-tartrate precipitate
3. Nylander’s test Bismuth subnitrate Black-colored solution
Bi(OH) NO + KOH Bi(OH) + KNO

2 3 3 3
2Bi(OH) + reducing sugar + heat 2Bi + O + 3H O
3 2
(black)
4. Barfoed’s test Cupric acetate in weak Red precipitate

acetic acid
2. Osazone formation
- Reducing sugars form characteristic osazone crystals when heated with an excess of
phenylhydrazine (C6H5NHNH2)
- Attributed to the presence of aldehyde or ketone group in their molecules
- Due to this property, sugars can be identified from a mixed sample
3. Action of alkalies
- Moore’s test – when a solution of reducing sugar is heated with an alkali (NaOH), it
turns yellow to orange, and finally dark brown liberating the odor or caramel, which
becomes more marked upon acidification
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4. Action of Acids
- monosaccharides when acted upon by concentrated acids boiling temperature are

dehydrated forming Furfural if the sugar is pentose; the sugar is a hexose,
Hydroxymethyl furfural (oxonium salts) is formed
A. Molisch test
Reagent: Alpha-naphthol and 95% ethyl alcohol and Conc. H2SO4
Positive Result: Violet ring at the junction of the two liquids
Use: General test for carbohydrates
B. Anthrone test
Reagent: Anthrone (Keto form of 9-hydroxyanthracene or 10-hydroanthracene-9-one) &
Conc. H2SO4
Positive Result: Blue or green color
Use: for rapid detection of carbohydrates in samples like body fluids and can be used for
cellulose assays
C. Seliwanoff’s Test
Reagent: Resorcinol and HCl
Positive Result: red color (specific for ketoses)
Use: used to differentiate aldoses from ketoses
D. Tollen’s Phloroglucinol test

Reagent: Phloroglucinol
Positive Result: Red color (for galactose)
Test for Pentoses
A. Bial’s Orcinol-HCl test

Reagent: Bial’s reagent: Orcinol in 95% Ethanol
10% FeCl3.6H2O
2 ml Orcinol-HCl (+ 1 ml sugar solution)
Positive Result: green solution to deep blue product
Use: To distinguish a pentose from a hexose
B. Tauber’s Benzidine Test

Reagent: 4% solution of benzedine in glacial acetic acid (tauber’s reagent)
1 ml Benzidine (+ 2 drops of sugar solution)
Positive result: Violet color
Use: test for the presence of pentoses
5. Fermentation
- decomposition of carbohydrates brought about by the action of microorganisms 9yeast,

molds, bacteria)). When yeast is added to sugar solution, ethyl alcohol and carbon
dioxide are produced
C6H12O6 = C2H5OH + CO2
- Zymase – enzyme in the yeast which produces fragmentation of the sugar molecule
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o This process in utilized in the manufacture of beverages and other industrial
products
6. Oxidation
- except sucrose, disaccharides and monosaccharides especially Aldoses are more

readily oxidized forming sugar acids
Oxidation of aldehyde group – forms “Aldonic acids”

Oxidation of Primary ROH – forms “Uronic acids”
Oxidation of both – forms “Aldaric acids”
7. Reduction
- all sugars except sucrose undergo reduction with the absorption of energy and the
formation of products convertible into fats
Reduction of an aldose produces one alcohol

Glucose + 2H = Sorbitol
Galactose + 2H = Dulcitol
Reduction of a ketose sugar yields two alcohols
Fructose + 2H = mannitol + Sorbitol
- these alcohols are used in culture media for the identification and differentiation of
bacteria
8. Esterification
- the presence of the primary alcohol group (-CH2OH) in the sugar molecule make it
reactive with an acid
DIGESTION AND ABSORPTION
- For complex carbohydrates to be of value, these should undergo digestion
60% of food ingested is complex carbohydrates

- Starch and glycogen – fragmented into simple monosaccharides
- Cellulose and pentosans – no digestive enzymes for humans
- Cellulose – cellulase – bacterial enzyme that acts on cellulose, however, nutritive value
is insignificant
- Pentosans – acted upon by bacteria to produce organic acids, alcohols and carbon
dioxide
Activity: name the sources of the following

Dietary carbohydrates:
- Starch: ________________________________________________________________
- Sucrose: _______________________________________________________________
- Cellulose: ______________________________________________________________
- Lactose: _______________________________________________________________
- Glycogen: ______________________________________________________________
- Pentosan: ______________________________________________________________
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Starch – the most common dietary form of carbohydrate
- Composed of:
o Amylose – linear pattern of glucose linked by α-1,4 glycosidic bonds
o Amylopectin – branched polymer; glucose linked by α-1,4 glycosidic bonds with
branching via α-1,6 - linkages
https://www.researchgate.net/figure/Chemical-structure-of-the-corn-starch-constituting-natural-polymers-amylopectin-
and_fig1_333944060
A. MOUTH/BUCCAL CAVITY
- First area where carbohydrates undergo digestion
- Contains Ptyalin or Salivary Amylase
Starch hydrolysis
- Hydrolysis of starch either by the action of amylolytic enzyme or by acid gives

rise to split fragments with simultaneous production of maltose
- In the presence of maltase, maltose is converted to glucose. In acid, however,
the final product is glucose
- Starch amylase maltose maltase glucose
- Starch acid Glucose
2 classes of amylases
- Alpha – amylases (alpha-1,4 glucan 4 glucanohydrolase)
• Found in pancreatic and salivary juices
• Saccharogenic amylase
• Found in human GIT
• Split alpha 1,4 glycosidic bonds (except those of maltose) in random
fashion
- Beta-amylases (alpha-1,4 glucan maltohydrolase)
• E.g barley malt
• Dextrinogenic amylase
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• Breaks β-1,4 glycosidic linkage
• Not found in human GIT
• Equivalent enzyme is CELLULASE ; found among ruminants or grass-
grazing animals with large cecum (e.g. cows, carabaos, horses, goats)
• Effects the successive removal of disaccharides (maltose units) from the
non-reducing ends
o The enzyme cannot hydrolyze the alpha-1,6 bonds and the
reaction stops at this point.
o The fragment which remains is called DEXTRIN or LIMIT
DEXTRIN
B. STOMACH
- Little digestion of polysaccharide
- Gastric juice has no carbohydrate-splitting enzyme
- Salivary amylase is inactivated by pepsin
- Fructosans are broken down by HCl
C. SMALL INTESTINES
- Digestion of polysaccharides & disaccharides is complete
- pancreatic amylase and disaccharidases hydrolyze CHO into monosaccharides
- CHO absorbed in the jejunum in the form of monosaccharides
- Undigestible CHO due to β-1,4 glycosidic bonds
- Examples: high-fiber fruits and vegetables which contain Cellulose, hemicelluloses and
Pectin https://www.slideshare.net/soniherat/digestion-and-absorption-of-carbohydrate
- In the L.I., these form bulk or
roughage which swells with water,
promoting peristalsis and easier
evacuation of stool.
ABSORPTION
- Pores of the mucosa through which
diffusion occurs are impermeable to
water soluble solutes with MW > than
100.
- Simple Diffusion: pentose
- Facilitated Diffusion: involves a
carrier protein or lipoprotein (fructose
and mannose)
- Active Transport: glucose and
galactose
- Secondary active transport - A
carrier transport is present in the
brush border of the epithelial cell.
o The carrier has a receptor site
for both glucose and sodium.
▪ It will not transport
glucose to the inside of
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the cell if the receptor sites for both glucose and sodium ion are not
simultaneously filled.
▪ The energy to cause the movement of the carrier from the exterior of the
membrane to the interior is derived from the difference in sodium
concentration between the outside and the inside. As sodium diffuses to
the inside of the cell, it drags the carrier and glucose along with it, thus
providing the energy for the transport of glucose.
Relative rate of transport of monosaccharides (glucose as standard)

- Galactose 110%
- Glucose 100%
- Fructose 43%
- Mannose 19%
- Pentoses 9%
* The absorbed monosaccharides are brought to the 3 big veins of the GIT. 1 fuses with 2, 2
fuses with 3 to form a big portal vein to transport monosaccharides into the LIVER
VEINS of the GIT
1. Superior mesenteric veins

2. Splenic veins
3. Inferior mesenteric veins
PORTAL VEIN Liver
Factors affecting absorption of CHO

1. Glucose decreases the absorption rate of galactose and xylose
2. Galactose decreases the absorption rate of glucose but no effect on xylose
3. Phlorhizin (structurally similar to glucose) inhibits glucose transport
a. Causes phlorhizin glycosuria
4. Vitamins: thyroxine and the B complex vitamins (thiamine, pyridoxine, pantothenic acid)
promote absorption of hexoses
5. Hyperglycemia – increased sugar levels in the blood
a. Results when the liver cannot completely clear the CHO from the blood and it
goes to the systemic circulation
b. Happens after 1 hour of ingesting large amounts of CHO
c. Goes back to normal after 2 hours or even to fasting level
d. This is the basis for the glucose tolerance test
6. Sprue – passage of carbohydrates (and lipids) in intestinal mucosal cells is inhibited
a. Oral administration of CHO do not follow the normal GTT but IV administration
follow normal GTT curve
b. Poisoning the GIT with iodoacetate and phlorhizin produces the same result
7. Liver and muscle converts glucose to glycogen
_________________________1. Substance responsible for the brick-red precipitate for
Benedict’s test and Fehling’s test.
_________________________2. Structure in carbohydrates responsible for the reducing
properties of carbohydrates.
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_________________________3. Which of the disaccharides do not have reducing properties?
_________________________4. Which group of carbohydrates is absorbed using simple
diffusion?
_________________________5. Which of the carbohydrates form bulk or roughage in the large
intestines?
Scoring Rubrik for Short Response Assignment and Case Analysis
4 3 2 1
unsuccessful
accuracy
correctly cited
slows readability
ASSIGNMENT:
1. Explain how the glucose tolerance test is performed and include the principle behind it.
2. Why would a person with sprue not follow the normal GTT curve when administered
orally as opposed to IV administration?
3. Accomplish all activities in this unit.
1. Starch blockers had been marketed many years ago as a means of losing weight without
having to exercise or reduce your daily caloric intake. Starch blockers were based on a
protein found in beans
a. What specific enzyme is blocked by starch blockers?
Thus, as the advertisements proclaimed, one could eat a large amount of starch during a
meal, and as long as you took the starch blocker, the starch would pass through the
digestive tract without being metabolized. Unfortunately, this was too good to be true,
and starch blockers were never shown to be effective in aiding weight loss.
b. Name some factors that make starch blockers too good to be true for aiding in
weight loss
2. Beans, peas, soybeans and other leguminous plants contain oligosaccharides with (1,6)-
linked galactose residues that cannot be hydrolyzed for absorption, including sucrose
with 1,2, or 3 galactose residues attached. What is the fate of these polysaccharides in
the intestines?
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LABORATORY:
INTRODUCTION
Many monosaccharides such as glucose and fructose are reducing sugars, meaning that they
possess free aldehyde (-CHO) or ketone (-C=0) groups that reduce weak oxidizing agents such
as the copper in Benedict's reagent.
Benedict's reagent contains cupric (copper) ion complexes with Figure 2. Glucose, a
monosaccharide and Sucrose, a disaccharide.
Benedict's test identifies reducing sugars based on their ability to reduce the cupric (Cu2+) ions
to cuprous oxide at basic (high) pH. Cuprous oxide is green to reddish orange.
Name: ____________________________ Date: _____________________________

ACTIVITY NO. 12
THE REDUCING PROPERTIES OF CARBOHYDRATES
OBJECTIVES:
1. determine correctly which sugars have reducing property
2. correlate properly the reducing properties of sugars to certain reactions of
carbohydrates in the body
3. master completely the principles behind the tests and perform them with confidence
MATERIALS: Reference materials
Scoring rubric
For Procedures:
For Rationalization
not clear
not clear
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Fehling’s test: Watch: https://www.youtube.com/watch?v=yjLB_ntM324
1. Prepare 7 test tubes and label them appropriately with the sugars to be tested: glucose,
galactose, fructose, sucrose, lactose, starch and cellulose (cotton threads)
2. Meanwhile in another test tube, mix 3.5 mL of Fehling’s A and 3.5 mL of Fehling’s B
solution. Mix thoroughly. This should result to a royal blue colored solution. If not,
discard and ask for another set from the CSR.
3. Divide your resulting Fehling’s reagent and distribute them into 7 test tubes that were
previously labelled. 1mL for each tube.
4. Add 0.5 mL of the appropriate carbohydrate to the corresponding tubes and mix
thoroughly
5. Place them in the boiling water bath for 3 minutes (or until a color change is observed)
6. Write your observations in the table below
Benedict’s test: Watch: https://www.youtube.com/watch?v=PAKCgrnKeBA
1. Prepare 7 appropriately labelled test tubes

2. Add 0.5 mL of the same sugars used in the previous test
3. Add 0.5 mL of Benedict’s reagent and mix thoroughly
4. Place them in the boiling water bath for 2 minutes
5. After the required time, remove from the water bath and write your observations on the
table below.
Tollen’s test: https://www.youtube.com/watch?v=cZ9IW6mxLCY
1. Prepare 7 appropriately labelled test tubes. Make sure that these test tubes are clean
and free from contamination
2. Add 0.5 mL of the same sugars used in Fehling’s test
3. Add 1 mL of Tollen’s reagent and mix gently
4. Place in a boiling water bath for 3-5 minutes
5. Note all the changes in the test tubes and write your observations below.
Barfoed’s test: Watch: https://www.youtube.com/watch?v=yQfMqvOxPrc
1. Prepare 7 test tubes and label them appropriately

2. Place 1.5 mL of Barfoed’s reagent into the test tubes
3. Add 8 drops of the same carbohydrates used in Fehling’s test and mix thoroughly
4. Place them in the boiling water bath for 2 minutes or more
5. Remove and allow to cool
6. Observe each tube and write your observations below
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Observations:
Carbohydrate Fehling’s Benedict’s Tollen’s Barfoed’s Interpretation
➢ Which group of carbohydrates showed reducing properties?

___________________________________________________________________
➢ Which of the test above gave a different set of results?
___________________________________________________________________
➢ Why is it different?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
_______________________________________________________
➢ Make a generalization about the reducing properties of carbohydrates.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
NB: Download a photo of the positive results for each test and attach it below each procedure.
Caption your photos properly.
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Scoring Rubric
1. What is a reducing sugar?

2. State the principle of the tests performed in this activity
3. Between Fehling‘s solution & Benedict’s solution, which is the better Reagent for clinical
tests and why? Explain briefly.
4. An increased sugar in your body is a manifestation of what disease? Give its signs and
symptoms.
5. What would be the best application for Barfoed’s test?
6. What is the importance of knowing the reducing properties of sugars?
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LABORATORY
INTRODUCTION
Sucrose is one of the sweetest sugars, surpassed only by fructose an invert sugar. It is
soluble in water, dextrorotatory and does not reduce mild oxidizing agents.
When hydrolyzed it becomes levorotatory due to the fact that the fructose has greater
levorotation than glucose has dextro-rotation. Because of this inversion of the specific rotation,
the hydrolysis of sucrose is known as inversion and the product of sucrose hydrolysis is called
invert sugar
In plants, glucose and fructose are involved in signaling pathways in which sucrose
concentration functions as a key sensor of the nutritional status of plants. Therefore, invertase
plays a key role in the control of cell differentiation and development.
Humans show a marked preference for diets containing sucrose, however, our genomes do not
code for invertase. Instead, we use a different and unrelated enzyme to hydrolyze sucrose,
sucrose-glucosidase (EC 3.2.1.48).
Name: ____________________________ Date: _____________________________

ACTIVITY NO. 13
INVERSION OF SUCROSE
OBJECTIVES

1. understand completely how sucrose is hydrolyzed inside and outside the body
2. write properly the word equation for the hydrolysis of sucrose
3. design accurately a method for testing the products of hydrolysis of sucrose
Scoring rubric
For Procedures:
For Rationalization
For Equations
- 3 points is given for every equation made
o 3 – all parts of the equation was written and the proper formula was used
o 2 – 1 mistake was identified by the instructor
o 1 – 2 mistakes was identified by the instructor
o 0 – no equation was written
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not clear
not clear
Use your reference materials to answer the following procedures.
➢ Recall the reducing properties of sucrose. Does sucrose have reducing properties?
_________________Why? _____________________________________________
______________________________________________________________________
________________________________________________________________
Hydrolysis of Sucrose
1. Place 6 mL of 5% sucrose in a 16x150 test tube
2. Add 2 drops of concentrated HCl and mix
3. Heat in a water bath for 15 minutes
4. Remove from heat
Write the word equation for the acid hydrolysis of sucrose
___________________________________________________________________
Neutralization
1. Get 1 mL from the hydrolyzed solution and place it in a test tube
2. Place a red litmus paper in it
3. Add 10% NaOH drop by drop until the red litmus paper turns to blue
4. Fish out the litmus paper
5. This solution is now neutralized. If it is cloudy, you may filter it.
Why is there a need to neutralize the hydrolysate before proceeding with the test?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Test for the hydrolysate
1. Based on your equation on the hydrolysis of sucrose, what test/s can you possibly use to
detect the presence of the products?
______________________________________________________________________
______________________________________________________________________
2. Why did you choose these tests?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
3. Based on your assumptions, state your hypothesis
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
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4. Proceed with your test/s but use your neutralized solution, follow the proportions used in
previous experiments and write your observations below
Carbohydrate Test used Result Conclusion/

tested for interpretation
5. Is there a difference in the reducing property of sucrose before and after hydrolysis?
___________________________________________________________________
6. Explain the difference in results
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
*Download a photo of the positive result of the test you have chosen and attach it below this
instruction. Make sure to label your work properly.
CONCLUSIONS
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
Scoring Rubric
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1. What is an invert sugar?
2. Why is the hydrolysis of sucrose called inversion?
3. Use word formulas to illustrate the chemical equation for the hydrolysis of:
a. Lactose
b. Maltose
4. Compare sucrose and invert sugar as to their relative sweetness, reducing property,
Seliwanoff’s test and optical property. Tabulate your answer.
5. If a person wants to lose weight is it advisable to totally eliminate sugar in your dietary
intake? Why?
6. In the body, how is sucrose hydrolyzed? What biochemical pathways will the products
of sucrose hydrolysis enter in order to be used as source of energy?
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Week no 9
Submodule no. 9
Carbohydrate
metabolism
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LESSON: Carbohydrate Metabolism
TOPICS:
6.6.1. Glycolysis
6.6.2. Kreb’s cycle
6.6.3. Electron Transport Chain
LECTURE: 6 hours
LABORATORY: 3 hours
1. Discuss completely the different reactions involved in the metabolism of glucose
2. Write correctly the different steps involved in glycolysis
3. Appreciate thoroughly the significance of the metabolism of different biomolecules in
the maintenance of normal life processes;
4. understand completely the principles used in the laboratory activities
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LESSON PROPER
INTRODUCTION
Glucose is the universal fuel of human cells. Every cell type in the human body is able to
generate adenosine triphosphate (ATP) from glycolysis, the pathway in which glucose is
oxidized and cleaved to form pyruvate. The importance of glycolysis in our fuel economy is
related to the availability of glucose in the blood, as well as the ability of glycolysis to generate
ATP in both the presence or absence of oxygen. Glucose is the major sugar in our diet and the
sugar that circulates in the blood to ensure that all cells have a continuous fuel supply. The
brain uses glucose almost exclusively as fuel.
Why complex reactions?
Metabolic intermediates
- Used in other metabolic processes in the cell
- Multiple step pathway helps the cell to handle metabolic energy efficiently
- The quantity of energy released in each step is small enough to be handled by the cell
* All cellular processes are

interrelated
Claude Bernard
- Demonstrated that blood sugar levels vary in the different areas of the circulation
o Portal blood – less sugar
o Hepatic vein – more sugar
▪ There must be some regulating mechanism in the liver
CHO malabsorption/dissacharide intolerance

- Lactose intolerance/ lactase deficiency syndrome
- Sucrose intolerance
- a-dextrinase deficiency (infants and children)
Incomplete digestion of amylopectin, maltose, sucrose or lactose leads

to bacterial decomposition in the LI
o Incomplete digestion of amylopectin, maltose, sucrose or lactose leads to

bacterial decomposition in the LI
- Leads to the production of glucose and other monosaccharides + organic acids and
gases
- Produces flatus and diarrhea
Blood Glucose
NV = 80 – 100 mg%
FBS = 70 – 100 mg%
Postprandial = 130 – 160 mg%
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- The CNS is dependent on these normal values
- Glucose is the major energy source that crosses the blood-brain barrier
- If glucose falls to 25 mg%
o Person becomes stuporous
o After 10 minutes = coma
o Irreversible after 30 minutes
- The muscle can derive energy from ketone bodies
- The myocardium take up fatty acids and lactic acid from blood and use as source of
energy
o The heart is insensitive to fluctuations of blood sugar levels
Physiological and hormonal

mechanisms keeps blood sugar
levels at 60 mg% even in
prolonged starvation
Factors that lower blood glucose levels
1. Glycogenesis – transformation of glucose into glycogen

2. Glycolysis – utilization of glucose for heat and energy
3. Lipogenesis – conversion of glucose to fats
4. Formation of certain amino acids
5. Glucosuria – excretion of glucose in the urine
Hormone that lowers blood glucose levels

1. Insulin
- Hormone secreted by B-cells of islets of Langerhans in pancreas
- Produces hypoglycemic effect by
o Increasing glycogenesis
o Decreasing gluconeogenesis
o Increasing glucose utilization by tissues
o Promoting lipogenesis
- Facilitates entrance of glucose in the cell where it is immediately phosphorylated to be
converted to energy to undergo glycolysis
- Deficient insulin diminishes entrance of glucose in the cell, thus less phosphorylation
- Glucose remains in the blood producing hyperglycemia
- If its concentration exceeds the renal threshold, it is excreted in the urine
- This is known clinically as diabetes mellitus
To cover energy requirements: (during hypoglycemic state)

1. Gluconeogenesis
- Production of glucose from non-carbohydrate sources like proteins
- will increase nitrogen excretion in the urine
- Wasting of body tissues
2. Lipolysis
- Breakdown of fats
- Happens when the patient losses 100 – 200 g of glucose per day
- Leads to ketonemia
o When aceto acetyl CoA is produced in excess during catabolism of fats for the
capacity of the extrahepatic tissues to utilize, they accumulate in the blood
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- Complication
o Renal excretion of acetoacetic acid and B-hydroxybutyric acid in the form of Na
salts
o The blood is depleted of Na producing acidosis
o Large quantity of fluid is lost = dehydration
Factors that increase blood glucose level
1. Glycogenolysis
- Conversion of liver glycogen into blood glucose
2. Gluconeogenesis
- Synthesis of blood glucose from non-CHO sources
Hormones that increase blood Glucose
1. ACTH both have diabetogenic effect

2. TSH
3. Adrenal cortical hormones
- 11-oxysteroid groups – with anti-insulin activity
o Corticosterone (Compound B)
o 11-dehydro-17-hydroxycorticosterone (compound E, cortisone)
o 17-hydroxycorticosterone (compound F, hydroxycortison, cortisol)
4. Thyroxine or tetraiodothyronine
- Increase absorption of hexoses from intestines
- Stimulates liver glycogenolysis
5. Glucagon – from A cells of islets of Langerhans
- promotes hepatic glycogenolysis and gluconeogenesis
6. Epinephrine and norepinephrine (catecholamines)
- promotes both liver and skeletal glycogenolysis
- physical or emotional stress cause increase production of epinephrine and an immediate
production of glucose for energy
7. Somatostatin: (delta cells)
- It inhibits secretion of insulin and glucagon
- also inhibits the release of GH
8. Growth hormone / Somatotrophic hormone: (anterior pituitary gland)
- antagonist to insulin;
- it stimulates lipolysis
9. Human Placental Lactogen (hPL)/Human Chorionic Somatomammotropin
- with anti-insulin activity
- gestational diabetes
Immediate Effects in Increased BGl

- Physiologic increase in the uptake of glucose by liver and brain cells
- Release of insulin
- Increase uptake of glucose by peripheral tissues
- Inhibition of glucagon release
Metabolic Pathways:
Activity:
Completely Define the following:
1. Glycolysis
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2. Glucogenesis
3. Glycogenesis
4. Glycogenolysis
5. Glyconeogenesis
6. Pentose-Phosphate pathway/Hexose Monophosphate shunt
Fate of Glucose
1. Blood glucose (also called physiologic sugar)
2. Glycogen - stored glucose
3. Alpha-keto acids – part of non-essential amino acids
4. Constituent of body structures
5. Lipogenesis – produce adipose fat (from glycerol)
6. Pyruvate – key metabolite in the
Kreb’s cycle
Steps in ATP production from glucose

molecules-- Overview
Glycolysis
Oxidative decarboxylation
Krebs cycle
Oxidative phosphorylation
Glucose – 6 – PO4 (product after

phosphorylation of glucose)
- Udergo glycolysis (anaerobic) via EMP
- Enter Kreb’s cycle (aerobic)
- Enter HMS (Hexose Monophosphate shunt)
- Converted to glycogen
- Converted to fat
- Excreted through the urine
GLYCOLYSIS
- Catabolism of glucose
2 types
1. Aerobic
- In the presence of oxygen
- Produces pyruvic acid, carbon dioxide, water and heat
2. Anaerobic
- In the absence of oxygen
- Produces lactic acid and energy
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2 Major Pathways:
Embden-Meyerhof pathway (EMP)

- Also known as the glycolytic pathway
- Glucose is broken down to pyruvic and lactic acid in the absence of oxygen (anaerobic)
Kreb’s Cycle
- Acetyl CoA (EMP)is split to carbon dioxide, water and energy in ATP
- This is the final common pathway for glucose, fatty acids and certain amino acids
- 90% of energy is derived from this pathway
EMBDEN-MEYERHOF PATHWAY (EMP)

- Also known as the glycolytic pathway
- Glucose is broken down to pyruvic and lactic acid in the absence of oxygen (anaerobic)
- The central pathway for glucose catabolism
2 Stages
- Phosphorylation and cleavage of glucose
o End product: glyceraldehyde-3-PO4
- Conversion of glyceraldehyde-3-PO4 into lactic acid, with energy yield conserved as
ATP
I. Phosphorylation and Cleavage of Glucose

1. Glucose is phosphorylated to glucose-6-PO4
- Enzyme: Hexokinase and Glucokinase
o Hexokinase
▪ Also catalyzes phosphorylation of fructose, mannose and glucosamine
aside from glucose
▪ Has higher affinity for aldohexoses than ketohexoses
o Glucokinase
▪ Phosphorylate only glucose
▪ Has lower affinity for glucose than hexokinase
2. Conversion of glucose-6-PO4 to fructose-6-PO4
- Enzyme: phosphoglucoisomerase
3. Phosphorylation of fructose-6-PO4 to fructose-1,6 diPO4
- Enzyme: phosphofructokinase
- A second molecule of ATP is invested to phosphorylate in position 1
- The reaction indicates that it is essentially irreversible in the cell
4. Cleavage of fructose 1,6 diPO4 to glyceraldehyder-3-PO4 and dihydroxyacetone PO4
- Enzyme: Aldolase
5. The interconversion of triose PO4
- Only glyceraldehyde-3-PO4 can be degraded in the further reaction of glycolysis
- DHA-PO4 is reversibly converted to glyceraldehyde-3-PO4
- Enzyme: Triose PO4 isomerase
- One molecule of glucose forms 2-glyceraldehyde-3-PO4
II. Conversion of Glyceraldehyde-3-PO4 into lactic acid
1. Oxidation of glyceraldehyde-3-PO4 to 1,3 diphosphoglycerate

Enzyme: glyceraldehyde dehydrogenase
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2. Transfer of one phosphate group of 1,3 diphosphoglyceric acid to ADP forming 3-
phosphoglyceric acid and ATP
Enzyme: phosphoglyceric acid kinase
3. Conversion of 3-phosphoglyceric acid to 2-phosphoglyceric acid

Enzyme: phosphoglucomutase
4. Dehydration of 2-phosphoglyceric acid to phosphoenol pyruvic acid

Enzyme: enolase
5. Transfer of phosphate group from phosphopyruvic acid to ADP

Enzyme: pyruvic acid phosphokinase
6. Reduction of pyruvic acid to lactic acid

Enzyme: lactic acid dehydrogenase
- Lactic acid is the end product of glycolytic sequence under anaerobic conditions and
lactic acid is a blind alley in metabolism
- Lactic acid once formed is not further utilized
- Fatigue and rigor of muscle fibers is in part due to acidification by lactic acid
Net synthesis of 2 molecules of ATP

for every molecule of glucose
converted to lactic acid
Why only 2 ATP’s?
Inhibition of Anaerobic Glycolysis by Oxygen (Pasteur Effect)

- Rate of glycolysis was based on the disappearance of glucose and accumulation of
lactic acid
- The rate of glycolysis is slowed in the presence of Oxygen
- Oxygen: lactic acid CO2 and H2O
- Energy released: utilized to resynthesize glucose from remaining lactic acid
- Net Effect:
o Decrease in the total consumption of glucose
o Less accumulation of lactic acid
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THE EMBDEN-MEYERHOFF PATHWAY
http://medical-dictionary.thefreedictionary.com/_/viewer.aspx?path=dorland&name=pathway_Embden-Meyerhof.jpg
DISEASES ASSOCIATED WITH IMPAIRED GLYCOLYSIS
HEXOKINASE DEFICIENCY
- In patients with inherited defects of hexokinase activity, the red blood cells contain low
concentration of the glycolytic intermediates, including the precursor, 2,3-
Diphosphoglycerate
- In consequence, the Hemoglobin of these patients has an abnormally high oxygen
affinity
- The oxygen saturation curves of RBCs from a patient with hexokinase deficiency are
shifted from the left, which indicates that Oxygen is less available for the tissues
PYRUVATE KINASE DEFICIENCY (Hemolytic Anemia)

- All RBCs are completely dependent upon glycolytic activity for ATP production
- Failure of the pyruvate kinase reaction drastically impedes the production of ATP
- Inadequate production of ATP reduces the activity of the Na+ and K+ stimulated
Adenosine Triphosphatase ion pump, which maintains the shape of the RBC membrane.
In consequence, the cells swell and lyse, and this excess RBC destruction results in
hemolytic anemia
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LACTIC ACIDOSIS
- Blood levels of Lactic acid are normally less than 1.2mM. In lactic acidosis, the values
for blood lactate may be 5.0 mM or more
- The high concentration of lactate results in lowered blood pH and bicarbonate levels
- High blood lactate levels can result from increased formation or decreased utilization of
lactate
- A common cause of hyperlactidemia is ANOXIA, in which:
o The formation of lactate is increased, because the shortage of oxygen reduces
mithochondrial production of ATP with the consequent activation of PFK, causing
increased glycolysis and lactate production
o The use of lactate by the tissues is reduced because the use of lactate as an
energy source requires oxygen
- Tissue Anoxia may occur in shock and other conditions that impair blood flow, in
respiratory disorders, and in severe anemia.
FATE OF PYRUVATE
- Reconverted to glucose-6-phosphate
- Combine with NH3 to form alanine
- May combine with CO2 to form oxaloacetic acid
- May be oxidatively decarboxylated and combine with CoA to form acetyl CoA
FATE OF LACTATE
- Circulated back to the liver and converted to glycogen
- Converted to pyruvate to enter Kreb’s cycle
https://www.researchgate.net/figure/Cori-cycle-and-glucose-alanine-cycle-These-are-the-cycles-that-link-glucose-
production_fig5_236914625
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A simplified Diagram showing acetyl-CoA is at the hub of protein, carbohydrate and fat
metabolism
Aerobic Respiration
- process where the pyruvate produced in glycolysis undergoes further breakdown.
- requires oxygen and yields much more energy than glycolysis.
- Two processes:
o Krebs cycle
▪ also known as the citric acid cycle, or the Krebs cycle, after Hans Adolf
Krebs who identified the cycle.
▪ a series of chemical reactions of central importance in all living cells that
use oxygen as part of cellular respiration.
▪ part of a metabolic pathway involved in the chemical conversion of
carbohydrates, fats and proteins into carbon dioxide and water to
generate a form of usable energy.
o Electron Transport Chain and Oxidative Phosphorylation
▪ produces ATP through chemiosmotic phosphorylation.
TRICARBOXYLIC ACID CYCLE/CAC/KREB’S CYCLE

- Final common pathway of oxidative catabolism of all fuel molecules in aerobic cells
(mitochondrial matrix)
- Greater amount of energy is released when glucose is completely oxidized to CO2 and
H2O
o This is because the product of glycolysis, the lactic acid is still complex while the
product of respiration, CO2 is a smaller molecule whose carbon atom is
completely oxidized.
o H ion is then fed into the respiratory chain, a series of electron carriers
- Also known as Kreb’s cyle, citric acid cycle
- First described by Hans Adolf Kreb (1900-1981)
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- Occurs in the mitochondria
- Starts with acetyl group from carbohydrates, lipids and amino acids
- Common pathway of oxidative catabolism of all fuel molecules in aerobic cells
- AMPHIBOLIC pathway – involves both anabolic & catabolic processes
o ANABOLIC process – provides routes for the synthesis of biosynthetic
precursors (e.g. α-Ketoglutarate, Oxaloacetate, Succinyl CoA)
o CATABOLIC process – degradation of 2-C acetyl residues from CHO, fatty
acids & amino acids
- Starts with acetyl group from carbohydrates, lipids and amino acids
- Common pathway of oxidative catabolism of all fuel molecules in aerobic cells
Pyruvate
- Link between EMP and TCA
- Does not directly enter TCA
- Converted to acetyl CoA
Conversion of pyruvic acid to acetyl CoA
3 steps involved
1. Decarboxylation
o Removal of C from pyruvic acid and released as CO2
2. Oxidation
o Removal of H+ and accepted by NAD+
3. Condensation of acetic acid with CoA to produce acetyl CoA
https://sites.google.com/site/metabolicprocesshersi/poll/cellular-respiration/oxidative-decarboxylation-krebs-cycle
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Individual Reactions in the TCA
Step 1: Condensation
- Transfer of 2-C acetyl group to the
oxaloacetate a 4-C compound to
form 6-C citrate
- Enzyme: Citric acid synthetase
o Regulatory enzyme
o Inhibited by ATP
- Methyl group of acetyl CoA
condenses with the carbonyl atom of
oxaloacetic acid with hydrolysis
thioester bond and formation of free
CoA
Citrate
- If it accumulates in the mitochondria
o Inhibitory to phosphofructokinase
o Slows glucose metabolism by way of glycolysis
- If it accumulates in the cytoplasm
o Results in citrate cleavage thru citrate lyase
Step 2: Citrate is rearranged to form

isocitrate with the temporary formation of cis-
aconitate
- Enzyme: Aconitase
Step 3: Decarboxylation
- Isocitrate (6-C) is oxidized and CO2 is removed to produce α-ketoglutarate (5-C)
All dehydrogenase reactions make

NADH or FADH2
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2 steps
o Oxidative decarboxylation of a-
ketoglutaric acid to succinyl CoA
▪ Enzyme: a-ketoglutaric acid
synthetase
o Succinyl CoA undergoes loss of its CoA
▪ Enzyme: succinyl thiokinase or
succinyl CoA synthetase
Step 5: CoA is removed from succinyl

CoA to produce succinate
- Coupled with the synthesis of GTP
- GTP is very similar to ATP
- GTP donates its terminal phosphate
group to ADP to form ATP
Step 6: Oxidation
- Oxidation of succinic acid to fumaric acid
- Enzyme: flavoprotein succinate dehydrogenase
(w/ FAD)
- FADH2 can donate electrons to various electron
acceptors
Step 7: Hydration
- Water is added to fumarate (4-C) producing malate (4-C)
- Enzyme: fumarase
This reaction involves addition of

water across the double bond
Step 8: Oxidation
- Malate is oxidized to form oxaloacetate
- Enzyme: malic acid dehydrogenase (w/ NAD)
- This reaction makes NADH
- It regenerates oxaloacetate for another round.
https://www.kau.edu.sa/Files/0002526/files/20209_citric_acid%5B1%5D.pdf
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Overall glucose tally
Glycolysis
2 ATP in 4 ATP = 2 ATP’s
2 NADH = 6ATP (or 4 ATP’s)
Kreb’s cycle
8 NADH (1 NADH will produce 3 ATP’s)
2 FADH2 (1 FADH2 will produce 2 ATP’s)
2 ATP’s = 2 ATP
Electron transport chain
3 x 8 NADH = 24 ATP’s
2 x 2 FADH2 = 4 ATP’s
Total: 36 or 38 ATP’s/glucose molecule
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The Kreb’s Cycle
Redox CoEnzymes
1. NAD+ - Nicotinamide adenine dinucleotide
2. NADH+ - Nicotinamide adenine dinucleotide (reduced)
3. FAD+ - Flavine adenine dinucleotide
- NAD+ and NADP+ are both from the vitamin niacin

- FAD+ is from the vitamin riboflavin
- For NAD+ and NADP+, nicotinamide is the active center, it recieves H
- For FAD+, flavin is the active site
- Main function: carry electrons to the mitochondrial electron transport chain
As the CoEnzymes are oxidized, molecular oxygen is reduced

2NADH + 2H+ + O2 MET 2NAD+ + 2H2O
2FADH2 + O2 MET 2FAD+ + 2H2O
*MET – Mitochondrial electron transport
Oxygen metabolism
Oxygen
- Final receptacle for electrons in the mitochondrial electron transport system
- Too much of it is dangerous to life
- Normal reaction:
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O2 + 4e- + 4H+ 2H2O
*water is a safe product

*other reduced products of oxygen
are dangerous
ROS – reactive oxygen species

- Can destroy many vital cell molecules
1. H2O2 – produced when redox reactions involve 2e-
O2 + 2e- + 2H+ H2O2
2. O2-1 – formed when redox reactions involve only 1e-
3. OH – hydroxyl radical - neutral OH that can react with almost anything in the cell
ROS toxicity
- Large bursts of ROS
o When skin is exposed to UV light = Cancer
o Stroke and heart attack
▪ Ischemia – limited oxygen supply (may be due to blood clot)
▪ Reperfusion – restoration of blood supply that cells cannot handle =
ROS bursts
Defense Mechanism against ROS

- Superoxide dismutase – destroys superoxide to form 1 hydrogen peroxide
2O2- + 2H+ H2O2 + O2
- Catalase – can convert H2O2 to H2O
2H2O2 O2 + H2O
High Energy Phosphate Bonds
- Phosphoanhydride bond
Phosphate + Phosphate-R + 35KJ diphosphate anhydride + H2O
▪ Both phosphate groups are negative radicals

▪ Repulsive force acts like spring
▪ Release of energy is through simple hydrolysis
ATP + H2O ADP + 35KJ

2 Types of Phosphorylation
Substrate level phosphorylation

- Process whereby energy derived from oxidation is used to form high energy PO4 bonds
on various biochemical molecules
- Examples: phosphoenol pyruvate, 1,3-diphosphoglycerate, phosphocreatine
- Phosphorylated substrate transfers phosphate to ADP for form ATP
- Happens under anaerobic conditions
Oxidative phosphorylation
- Process that directly uses energy from redox reactions to form ATP
- Occurs in the mitochondria
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- Depends on the electron transport system
- Needs oxygen to proceed, thus, aerobic
ASSIGNMENT:
- Glycolysis = EMP
C6H12O6 + 2ADP + 2Pi 2CH3CH(OH)COO- + 2ATP + 150KJ
- Complete oxidation of glucose = aerobic
C6H12O6 + 6O2 6O2 + 6H2O + 2820KJ
* If 1.7 mol of glucose is processed via EMP,

a. how many moles of pyruvate are produced?
b. how many moles of ATP are produced?
THE ELECTRON TRANSPORT CHAIN

1. The final common pathway in aerobic cells by which electrons derived from various
substrates are transferred to oxygen
2. many substrates are oxidized by enzymes that use NAD+ or FAD+ as electron acceptor
cofactors
The Mitochondrial Structure

3. Outer membrane – permeable
to most small molecules
4. Intermembrane space –
presents no barrier to
metabolites
5. Inner membrane – highly
selective; provides transport
systems for the following
substances:
o ATP, ADP & Pi
o Pyruvate, succinate, α-
ketoglutarate, malate &
citrate
o Cytidine and GTP
6. Note: The enzymes of the ETC
& oxidative phosphorylation,
including Succinate
dehydrogenase are found in the inner mitochondrial membrane
7. Matrix – contains the enzymes of the Kreb’s cycle (except Succinate dehydrogenase),
enzymes of β-oxidation of fatty acids and other miscellaneous enzyme systems
ORGANIZATION OF THE ELECTRON TRANSPORT CHAIN
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8. The cytochromes and the Fe-S protein centers are one-electron carriers.
9. NADH, FADH2 and Q are two-electron carriers
10. For every pair of electrons w/c come from NADH, 3 moles of ATP are generated
11. For every pair of electrons w/c come from FADH2, 2 moles of ATP are generated
SOURCES OF ELECTRONS
12. NADH is derived from NAD+-linked dehydrogenases, including:
o Isocitrate, α-ketoglutarate, and malate dehydrogenases of the TCA cycle
o Pyruvate dehydrogenase
o L-3-Hydroxyacyl CoA dehydrogenase of fatty acid oxidation
o Miscellaneous NAD+-linked dehydrogenase
13. FADH2 is derived from FAD-linked dehydrogenases, including:
o Succinate dehydrogenase of the TCA cycle
o FAD-linked dehydrogenase of the α-glycerophosphate shuttle
o Acyl CoA dehydrogenase of fatty acid oxidation
o Miscellaneous FAD-linked dehydrogenases
The electron transport chain is threefold:

1) to pass along 2H+ ions and 2e- to eventually react with oxygen;
2) to conserve energy by forming three ATP's; and
3) to regenerate the coenzymes back to their original form as oxidizing agents.
Initiation of Electron Transport Chain:

14. NADH interacts with the first complex 1 enzyme, known as NADH reductase.
15. This complex 1 contains a coenzyme flavin mononucleotide (FMN) which is similar to
FAD
*NADH + H+ enter the enzyme complex and pass along the 2
hydrogen ions to an interspace in the mitochondria.
*These hydrogen ions, acting as a pump, are utilized by ATP

synthetase to produce an ATP for every two hydrogen ions
produced
16. 2H+ = 1 ATP
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complexes 1, 3, 4 act in this manner to produce 2
hydrogen ions each, and thus will produce 3 ATP for
every use of the complete electron transport chain
NADH passes along 2 electrons to
(1) FMN
(2) iron-sulfur protein (FeS)
(3) to coenzyme Q.
- The net effect of these reactions are to regenerate coenzyme NAD+.

- This regeneration of reactants produces a cycling effect.
- The NAD+ is ready to react further with metabolites in the citric acid cycle.
(4) Coenzyme Q picks up an additional 2 hydrogen ions to make CoQH2

CoQ + H2 CoQH2
CoQH2 is soluble in the lipid membrane and can move through

the membrane to come into contact with enzyme complex 3.
(5) Coenzyme QH2 carrying an extra 2 electrons and 2 hydrogen ions starts a cascade of
events through enzyme complex 3, also known as cytochrome reductase b. (Enzyme complex
3)
- Cytochromes are very similar to the structure of

myoglobin or hemoglobin.
- The significant feature is the heme structure
containing the iron ions, initially in the +3 state and
changed to the +2 state by the addition of an electron.
(6) The CoQH2 passes along the 2 electrons to

(a) cytochrome b1 heme
(b) b2 heme ,
(c)iron-sulfur protein,
(d)cytochrome c1,
(e)finally to cytochrome c.
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(7) In the meantime the 2
hydrogen ions are channeled
to the interspace of the
mitochondria for ultimate
conversion into ATP.
http://chemistry.elmhurst.edu/vchembook/596electransport.html
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__________________________1. Name 2 sugars that are absorbed by facilitated transport
__________________________2.
__________________________3. Name 2 ions that are inhibitory to glucose absorption
__________________________4.
__________________________5. Enumerate the 4 steps in ATP production from 1 glucose
__________________________6. molecule in chronological order
__________________________7.
__________________________8.
__________________________9. Name the two one-electron carriers in the electron
__________________________10. transport chain
__________________________11. Name at least 2 hormones that is inhibited by somatostatin
__________________________12.
__________________________13. What bacterial enzyme is capable of fragmenting cellulose
into its component monosaccharides?
__________________________14. Which hormone can promote glycogenolysis and
gluconeogenesis?
__________________________15. Which part of the mitochondria presents no barrier to
metabolites?
__________________________16. Which enzyme in the Kreb’s cycle is found in the inner
mitochondrial membrane?
__________________________17. What is the key metabolite in the Kreb’s cycle?
__________________________18. What is the most significant feature of enzyme complex 3 of
the electron transport chain?
__________________________19. How many electrons is picked up by Coenzyme Q?
__________________________20. What is another name for enzyme complex 3?
ASSIGNMENT:
Scoring Rubrik for the short response assignment
4 3 2 1
unsuccessful
accuracy
correctly cited
slows readability
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1. Lactate and pyruvate are in equilibrium in the cell, and the ratio of lactate to pyruvate
reflects the NADH/NAD+ ratio. Both acids are released in to blood, and the normal ratio
of lactate to pyruvate in blood is approximately 25:1. This ratio can provide a useful
clinical diagnostic tool. Because lactic acidemia can be the result of a number of
problems, such as hypoxia, MERFF, thiamine deficiency and pyruvate dehydrogenate
deficiency, under which of these conditions would you expect the lactate/pyruvate ratio
in blood to be much greater than normal?
2. Answer the assigned tasks in this unit.
SUMMATIVE ASSESSMENT: highlight your answer. (2x)
1. A major role of glycolysis is which of the following?

A. To synthesize glucose D. To synthesize glycogen
B. To generate energy E. To use ATP to generate heat
C. To produce FAD (2H)
2. Starting with glyceraldehyde-3-phosphate and synthesizing one molecule of pyruvate,
the net yield of ATP and NADH would be which of the following?
A. 1 ATP, 1 NADH F. 2 ATP, 4 NADH
B. 1 ATP, 2 NADH G. 3 ATP, 1 NADH
C. 1 ATP’ 4 NADH H. 3 ATP, 1 NADH
D. 2 ATP, 1 NADH I. 3 ATP, 4 NADH
E. 2 ATP’ 2 NADH
3. When glycogen is degraded, glucose-1-phosphate is formed. Glucose –1- phosphate
can then be isomerized to glucose-6-phosphate. Starting with glucose-1-phosphate and
ending with 2 molecules of pyruvate, what is the net yield of glycolysis in terms of ATP
and NADH formed?
A. 1 ATP, 1 NADH F. 2 ATP, 3 NADH
B. 1 ATP, 2 NADH G. 3 ATP, 1 NADH
C. 1 ATP’ 3 NADH H. 3 ATP, 2 NADH
D. 2 ATP, 1 NADH I. 3 ATP, 3 NADH
E. 2 ATP’ 2 NADH
4. Which of the following statements correctly describes an aspect of glycolysis?
A. ATP is formed by oxidative phosphorylation
B. 2 ATP’s are used in the beginning of the pathway.
C. Pyruvate kinase is the rate-limiting enzyme
D. One pyruvate and 3 CO2 are formed from the oxidation of one glucose molecule
E. The reaction takes place in the matrix of the mitochondria
5. How many moles of ATP are generated by the complex aerobic oxidation of 1 mole of
glucose to 6 moles of CO2
A. 2 – 4 D. 30 – 32
B. 10 – 12 E. 60 – 64
C. 18 – 22
6. A patient with type I diabetes mellitus takes an insulin injection before eating dinner but
then gets distracted and does not eat. Approximately 3 hours later, the patient becomes
shaky, sweaty and confused. These symptoms have occurred because of which of the
following?
A. Increased glucagon release from the pancreas
B. Decreased glucagon release from the pancreas
C. High blood glucose levels
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D. Low blood glucose levels
E. Elevated ketone body levels
7. Which of the following organs have the highest demand for glucose as a fuel?
A. Brain D. Liver
B. Muscle (skeletal) E. Pancreas
C. Heart
8. An alcoholic patient developed a pancreatitis that affected his exocrine pancreatic
function. He exhibited discomfort after eating a high carbohydrate meal. The patient
most likely had a reduced ability to digest which of the following?
A. Starch D. Sucrose
B. Lactose E. maltose
C. Fiber
9. A type I diabetic neglects to take his insulin shots while on a weekend vacation. Cells of
which tissue would be most greatly affected by this mistake?
A. Brain D. Red blood cells
B. Liver E. Pancreas
C. Muscle
10. After digestion of a piece of cake that contains flour, milk and sucrose as its primary
ingredients, the major carbohydrate products entering the blood are which of the
following?
A. Glucose D. Fructose and glucose
B. Fructose and galactose E. Glucose, galactose and fructose
C. Galactose and glucose
11. A patient has a genetic defect that causes intestinal epithelial cells to produce
disaccharidases of much lower activity than normal. Compared with w normal person,
after eating a bowl of milk and oatmeal sweetened with table sugar, this patient will
exhibit higher levels of which of the following?
A. Maltose, sucrose, and lactose in the stool
B. Starch in the stool
C. Galactose and fructose in the blood
D. Glycogen in the muscles
E. Insulin the blood
12. Superoxide dismutase catalyzes which of the following reactions?
A. O2- + e- + 2H+ yields H2O2
B. 2O2- + 2H+ yields H2O2 + O2
C. O2- + HO + H+ yields CO2 + H2O
D. H2O2 + O2 yields 4H2O
E. O2- + H2O2 + H+ yields 2H2O + O2
13. A 25 y/o female presents with chronic fatigue. A series of blood tests are ordered, and
the results suggest that her red blood cell count is low because of iron deficiency
anemia. Such a deficiency would lead to fatigue because of which of the following?
A. Her decrease in FeS centers is impairing transfer of electrons in the electron
transport chain .
B. She is not producing as much H2O in the electron transport chain, leading to
dehydration, which has resulted in fatigue
C. Iron forms a chelate with NADH and FAD(2H) that is necessary for them to
donate their electrons to the ETC.
D. Iron acts as a cofactor for α-ketoglutarate DH in the TCA cycle, a reaction
required for the flow of electrons through the ETC
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E. Iron accompanies the protons that are pumped from the mitochondrial matrix to
the cytosolic side of the inner mitochondrial membrane. Without iron, the proton
gradient cannot be maintained to produce adequate ATP.
14. Coenzyme A is synthesized from which of the following vitamins?
A. Niacin C. Pantothenate
B. Riboflavin D. Vitamin C
C. Vitamin A
15. A patient diagnosed with thiamine deficiency exhibited fatigue and muscle cramps have
been related to an accumulation of metabolic acids. Which of the following metabolic
acids is most likely to accumulate in a thiamine deficiency?
A. Isocitric acid C. Malic acid
B. Pyruvic acid D. Oxaloacetic acid
C. Succinic acid
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LABORATORY
INTRODUCTION
Starch is a polysaccharide constructed from alpha D glucose that is linked by glycosidic

bonds. Polymers of glucose are called glucanes.
The starch molecule is made up of amylose molecules and amylopectin molecules.
Amylose molecules are largely unbranched water – soluble and contain mainly alpha –1,4
glycosidic linkages. They are like most polysaccharides, polydispers, meaning that the length of
the molecule is not exactly defined, the number of glucose units being between 200 to more
than 1000. The alpha – 1,4 linkages cause the helical structure of starch. The inner diameter of
helix is big enough for elementary iodine to be deposited forming a blue complex. Amylopectin
is characterized by branchings. The branches are via α-1,6 glycosidic linkages that are made up
of 12 glucose units and forms secondary helical structures which can also accommodate
elementary iodine in the inside forming a red – violet complex.
Glycogen is the storage form of glucose in animals and humans which is analogous to
the starch in plants. Glycogen is synthesized and stored mainly in the liver and the muscles.
Structurally, glycogen is very similar to amylopectin with alpha acetal linkages, however, it has
even more branching and more glucose units are present than in amylopectin. Various samples
of glycogen have been measured at 1,700-600,000 units of glucose. Glycogen uses alpha – 1, 4
glycosidic linkages and cross linked by alpha – 1,6 glycosidic linkages.
Plants make starch and cellulose through the photosynthesis processes. Animals and
human in turn eat plant materials and products. Digestion is a process of hydrolysis where the
starch is broken ultimately into the various monosaccharides. A major product is glucose which
can be used immediately for metabolism to make energy. The glucose that is not used
immediately is converted in the liver and muscles into glycogen for storage by the process of
glycogenesis. Any glucose in excess of the needs for energy and storage as glycogen is
converted to fat.
Name: _____________________________ Date: _____________________________

Activity No. 14
STARCH AND GLYCOGEN
OBJECTIVES

1. differentiate accurately the products of the acid hydrolysis and enzymatic hydrolysis of
starch
2. perform confidently a simple procedure to test for the products of the hydrolysis of
starch
3. recognize correctly the basic properties of starch
4. recognize accurately the basic properties of glycogen
5. differentiate correctly glycogen from starch
6. perform precisely simple tests to determine the properties of glycogen
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Scoring rubric
For Procedures:
For Rationalization
For Equations
- 3 points is given for every equation made
o 3 – all parts of the equation was written and the proper formula was used
o 2 – 1 mistake was identified by the instructor
o 1 – 2 mistakes was identified by the instructor
o 0 – no equation was written
o 3 – all concepts were mentioned and a clear hypothesis, generalization or conclusion was derived from the
activity
o 2 – one or 2 concepts were not mentioned and the hypothesis, generalization or conclusion is not clear
o 1 – 3 or more concepts were not mentioned and the hypothesis, generalization or conclusion is not clear
Properties of starch and glycogen
Reaction with Iodine

1. Recall how starch and glycogen reacts with iodine from previous activities
Starch: ___________________________________
Glycogen: ________________________________
2. What would be the color reaction of raw starch with iodine?

_____________________________
3. What would be the color reaction of cooked starch with iodine?
__________________________
4. Explain the difference between the color reaction of raw starch and cooked starch with
iodine?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Reducing properties of starch and glycogen

1. Recall the reducing properties of starch and glycogen from previous activities/lessons
Starch:
______________________________________________________________________
Glycogen:
_____________________________________________________________________
Hydrolysis of starch and glycogen

- The hydrolysis of starch and glycogen can be both actuated by acid and the enzyme
amylase.
- Write the complete word equation for the hydrolysis of starch in acid and with amylase.
Acid:
______________________________________________________________________
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Amylase:
_____________________________________________________________________
- Write the complete word equation for the hydrolysis of starch in acid and with amylase
Acid:
______________________________________________________________________
Amylase:
_____________________________________________________________________
Test for the products of hydrolysis

1. Based on your equation on the acid hydrolysis and with the enzymatic hydrolysis of
starch as well as glycogen glycogen, what possible test/s can you use to detect the
presence of these end product/s?
______________________________________________________________________
2. Why did you choose this test?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
3. From your assumptions, make a hypothesis.
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
4. Before testing for the presence of the end products of the hydrolysis of starch or
glycogen, whether acid or enzymatic, a process of neutralization should be done on your
test sample using either sodium bicarbonate or a few drops of sodium hydroxide.
Why is this so?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Is there a difference in the reducing properties of starch and glycogen before and after
hydrolysis?
______________________________________________________________________
Explain the difference.

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
❖ Download a photo of the positive result of the test you have chosen to test for the end
products of the hydrolysis of starch and glycogen. Attach it below this instruction.
Caption your work correctly.
CONCLUSIONS
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
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Scoring Rubric
1. Using flow charts, trace the acid hydrolysis of starch

2. Compare Amylose and Amylopectin as to their: size of molecule, molecular structure,
main mode of bonding, and reaction to iodine. Tabulate your answers
3. Differentiate the acid and enzymatic hydrolysis of starch.
4. Name other properties and uses of starch not mentioned in this activity
5. How is glycogen made in the body?
6. How is glycogen hydrolyzed in the body?
7. Compare the properties of Starch with that of Glycogen as to molecular structure and
reaction to iodine. Tabulate your answer
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Week no 10
Submodule no. 10
Metabolism of
other
carbohydrates
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LESSON: Metabolism of other Carbohydrates
TOPICS:
6.6.3. Gluconeogenesis
6.6.4. Glucogenesis
6.6.4.1. Fructose Metabolism
6.6.4.2. Galactose Metabolism
6.6.5. Pentose Phosphate Pathway
TIME /SCHEDULE: Week 10
LECTURE: 5 hours
LABORATORY: 4 hours
LEARNING OBJECTIVES

1. Describe completely the metabolism of other carbohydrates
2. Explain accurately the mode of actions of different hormones in the maintenance of
blood glucose levels;
3. Discuss completely the different processes involved in the maintenance of normal blood
glucose
4. Discern properly the importance of the knowledge of metabolism in keeping oneself
healthy
5. Appreciate thoroughly the significance of the metabolism off different biomolecules in the
maintenance of different life processes
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LESSON PROPER
INTRODUCTION
Carbohydrates are the largest source of dietary calories for most of the world’s
population. The major carbohydrates in the human diet are starch, lactose and sucrose. The
digestion of the disaccharides lactose and sucrose, as well as further digestion of maltose,
maltotriose and limit dextrins, occurs through disaccharidases attached to the membrane
surface of the brush border (microvilli) of intestinal epithelial cells.
Glycogen is the storage form of glucose found inmost types of cells. It is composed of
glucosyl units linked by α-1,4 glycosidic bonds, with α-1,6 branches occurring roughly every 8 –
10 glucosyl units. The liver and skeletal muscle contain the largest glycogen stores.
The formation of glycogen from glucose is an energy-requiring pathway that begin, like
most of the glucose metabolism, with the phosphorylation of glucose to glucose 6-phosphate.
Glycogen synthesis from glucose 6-phosphate involves the formation of uridine diphosphate
glucose (UDP-glucose) and the transfer of glucosyl units from UDP-glucose to the ends of the
glycogen chains by the enzyme glycogen synthase. Once the chains reach approximately 11
glucosyl units, a branching enzyme moves six to eight units to form an α (1,6) branch.
Glycogenolysis, the pathway for glycogen degradation, is not the reverse of the
biosynthetic pathway. The degradative enzyme glycogen phosphorylase removes glucosyl units
one at a time from the ends of the glycogen chains, converting them to glucose 1-phosphate
without resynthesizing UDP-glucose or UTP. A debranching enzyme removes the glucosyl
residues near each branchpoint.
Liver glycogen serves as a source of blood glucose. To generate glucose, the glucose 1-
phosphate produced from glycogen degradation is converted to glucose 6-phosphate. Glucose
6-phosphatase, an enzyme found only in liver and kidney, converts glucose 6-phosphate to free
glucose, which then enters the blood.
Metabolic Pathways:
Glycolysis: aka- Embden Meyerhof Pathway (glucose breakdown to form pyruvate and with
the release of ATP molecules as the source of biochemical energy.
Glucogenesis: conversion of non-glucose hexoses like mannose, fructose and galactose to

substrates found in the glycolytic pathway in order to form more ATP molecules.
Gluconeogenesis: conversion of non-carbohydrate sources like amino acids, glycerol and

lactate to substrates found in the glycolytic pathway in order to form more ATP molecules
Glycogenesis: synthesis of excess glucose molecules into ploymer called glycogen that will be
eventually stored in the liver. Regulated by the hormone INSULIN
Glycogenolysis: reverse process of glycogenesis. Breakdown of glycogen to glucose that will

be utilized for ATP production. Regulated by GLUCAGON
Pentose Phosphate Pathway/ Hexose Monophosphate Shunt/Phosphogluconate

Oxidative Pathway: Alternative pathway where glucose molecules can also be catabolized and
form ATP molecules.
Kreb’s Cycle/Citric Acid cycle/Tricarboxylic Acid cycle: It is the final common pathway for
the breakdown of foodstuff where acetyl coA is utilized to generate this cycle in producing ATP.
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GLUCONEOGENESIS
• conversion of non - carbohydrate sources like amino acids, glycerol and lactate to
substrates found in the glycolytic pathway.
- Involves removal and transfer of amino groups (transdeamination)
Q: WHAT CAN BE CONVERTED

BACK TO GLUCOSE?
A: Lactate, pyruvate, or any three-

carbon equivalent. Acetyl-CoA
CANNOT BE CONVERTED BACK
TO GLUCOSE!
LIVER: The main organ that does

gluconeogenesis.
MUSCLE: Muscle does

gluconeogenesis too if need be, to
create glucose-6-phosphate.
However, it cannot go all the way
back to free glucose.
Alanine + a-ketogluratarate ➔ pyruvate + glutamate
Aspartate + a-ketogluratarate ➔ oxaloacetate + glutamate
https://themedicalbiochemistrypage.org/gluconeogenesis-endogenous-glucose-synthesis/
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PATHWAY: Starting from Lactate, and going to Glucose
1. Lactate ------> Pyruvate (oxidation)

- Catalyzed by Lactate Dehydrogenase
- Concurrent NAD+ ------> NADH (reduction)
2. Pyruvate ------> Oxaloacetate

- The ultimate goal here is to get back to Phosphoenolpyruvate, so that we can - -continue
with the reverse of glycolysis. But since PEP is an irreversible reaction in glycolysis, we
need a workaround.
- This reaction catalyzed by Pyruvate Carboxylase.
- ATP is required.
- This reaction occurs in the mitochondria.
3. Oxaloacetate ------> Phosphoenolpyruvate

- GTP is consumed. So, in two steps we have gotten to PEP, at the expense of ATP and
GTP, or energetic equivalent of 2ATP.
- Catalyzed by Phosphoenolpyruvate Carboxykinase.
- The malate shuttle allows free passage of this intermediate to the cytosol, so that
gluconeogenesis can be completed.
4. Phosphoenolpyruvate ------> ------> Glucose-6-Phosphate

- The rest of the intermediates are the same as they are for glycolysis.
- Fructose-1,6-biphosphate ------> Fructose-6-Phosphate was irreversible in glycolysis, so
here it must be catalyzed by a different enzyme, although we still have the same
intermediates.
ENERGETICS OF GLUCONEOGENESIS: It
costs 6 ATP to make one glucose -- 4 ATP +
2 GTP.
LIVER ONLY: Glucose-6-Phosphatase can

catalyze Glucose-6-Phosphate ------> free
glucose, which can then be excreted into the
blood to raise blood-sugar. This does not
occur in muscles, although muscles undergo
gluconeogenesis.
REGULATION OF GLUCONEOGENESIS:
1. Positive Effectors: Generally things that indicate a surplus of energy.

- ATP
- Acetyl-CoA
- Citrate
2. Negative Effectors: Generally things that indicate a lack of energy, as well as glycolytic
intermediates.
- AMP
- Fructose-1,6-biphosphate or Fructose-2,6-biphosphate
- Inorganic phosphate
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Interconversion between Monosaccharides in the Liver
• Liver cells contain all appropriate enzymes needed to promote interconversions between
the monosaccharides
• Most human tissues cannot utilize galactose and fructose
– Galactose and fructose must be converted by the liver cells into glucose which is
then transported by the blood to the other cells
Fructose Metabolism
• Diets containing large amounts of sucrose can utilize the fructose as a major source of
energy
• Enzymes needed to metabolize fructose
– Fructokinase – phosphorylates fructose to fructose-1-P
– Hexokinase – phosphorylates fructose to fructose-6-P, this enzyme has much
higher affinity for glucose than fructose, thus the relative abundance of glucose in
the liver competitively inhibits the phosphorylation of fructose
– Aldolase – utilizes fructose-1,6 biphosphate as substrate
• The principal organs that can metabolize fructose are the liver, kidneys, intestines and
adipose tissue
Clinical significance of fructose metabolism
FRUCTOSURIA
- A benign disorder caused by the lack of fructokinase

- It is asymptomatic
- Harmless and may go undiagnosed
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- Affected individuals spill fructose into their urine which might be misinterpreted as
glucosuria, because both yield a positive reducing-sugar test
- Prolonged intake of fructose by infants with this defect leads to vomiting, poor feeding,
jaundice and eventually hepatic failure and death
Fructose Intolerance
- lack of aldolase (F-1,6 biPO4 cleavage)
- Char. by hypoglycemia and vomiting following sucrose or fructose intake.
- Hypoglycemia: Fructose-1-phosphate inhibits glycogenolysis and gluconeogenesis.
Aldolase: reversible cleavage of fructose ester into triose sugars

Fructose-1-P ……. Fructose-1,6 DiPO4….. Fructose-6-P ……. G-6-P
Patients remain symptom-free on a diet devoid of fructose
Galactose Metabolism
- Galactose is obtained from milk sugar, lactose

- The most important organs that can metabolize galactose are the liver and erythrocytes
- Galactosemia, an increased level of serum galactose, is a major symptom of two
enzyme defects, galactokinase and galactose-1-phosphate uridyl transferase
- Treatment: elimination of milk products as the source of lactose
- some capacity to metabolize ingested galactose may develop later in the life of a
galactosemic person by the development of the enzyme UDP Galactose phosphorylase
https://in.pinterest.com/pin/643944446690620192/
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- 2 enzyme defects: galactokinase and galactose-1-phosphate uridyl transferase
- In both disorders, galactose is reduced to its sugar alcohol, galactitol, which may be
deposited inside the lens of the eye. Osmotic pressure inside the lens increases and can
cause cataracts
- In only galactose-1-PO4
uridyl transferase deficiency,
galactose-1-PO4 is trapped
within the liver cells and
RBC, leading to
hepatomegaly, impaired
liver function and mental
retardation if the dietary
galactose intake continues
o The treatment for
both types of
galactosemia is to
eliminate milk
products, the source
of lactose, which
contains galactose
o Some capacity to metabolize
ingested galactose may develop
later in life of a galactosemic
person by the development of the
enzyme UDPgalpyrophosphorylase
Glycogenesis
- The synthesis of glycogen from monosassacharides

- All cells of the body are capable of storing at least some glycogen, but the liver cells and
muscle cells can store large amounts of glycogen
- This process makes it possible to store a large quantity of glucose without significantly
altering the osmotic pressure of the intracellular fluids
- Offers a relief of a hyperglycemic state. There is however a limit to the amount of
glycogen that can be stored in normal tissues. When this limit is exceeded, the excess
glucose is converted to fat, which has no limit as to its storage
- The glycogen store exists in two forms
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o Free glycogen – one that can be readily released
o Fixed glycogen – one that can be released only slowly
https://disorders.eyes.arizona.edu/disorders/galactokinase-deficiency
Glycogenolysis
- The breakdown of glucogen to reform glucose in the cells
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- Each succeeding glucose molecule on each branch of the glycogen polymer is split
away by a process of
phophorylation, catalyzed
by the enzyme
phosphorylase
- Phosporylase attacks the
glycogen molecule of
glucose residues, alpha
1,4 glucosidase bonds
until the branch points are
reached, then ceases, to
produce the limit dextrin
- Under resting conditions,
the phosphorylase is in an
inactive form so that
glucose can be stored in
the form of glycogen.
When it is necessary to re-
form glucose from
glycogen, glycogen must
first be activated
- Two hormones glucagon and epinephrine can activate phosphorylase and thereby
cause rapid glycogenolysis
- This process maintainins blood sugar. The cells of the liver, kidney tubules and small
intestine contain glucose-6-PO4 that can split phosphate away from G6P and therefore
make glucose available for retransport out of the cells into the intestinal fluids. Therefore
liver glycogenolysis causes an immediate rise in blood glucose concentration
- Glycogenolysis in most other cells of the body especially in the muscle cell but does not
release glucose into the extracellular fluids because G6P cannot be dephosphorylated
Glycogen Storage Diseases

- Genetically linked metabolic disorders that involve regulating glycogen metabolism
- Symptoms vary by the GSD type and can include muscle cramps and wasting, enlarged
liver and low blood sugar level.
- Glycogen serve as the primary fuel reserve for the body’s energy needs
- Disruption of glycogen metabolism will thus affect other biochemical pathways as the
body seeks alternative fuel sources
- Accumulation of abnormal metabolic products can damage the kidneys and other organs
A. Von Gierke’s Diases – GSD I

a. It is caused by G6P deficiency in the liver, kidney and small intestines, the last
step in glycogenolysis, the transformation of G6P to glucose does not occur
b. As a result the liver is clogged with excess glycogen and becomes enlarged and
fatty
c. Other symptoms include low blood sugar level and elevated levels of lactate,
lipids and uric acid in the blood
d. Growth is impaired, puberty is often delayed, and bones maybe weakened by
osteoporosis
B. Pompe’s Disease – GSD II
a. It is caused by lysososmal alpha D glucosidase deficiency in skeletal and heart
muscles
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b. GSD II in infants, the baby appears normal at birth, but within a few months they
develop muscle weakness, trouble in breathing and an enlarged heart
c. Cardiac failure and death usually occur before age 2 despite medical treatment
d. The juvenile and adult forms of GSD II affect mainly the skeletal muscles in the
body’s limbs and torso
e. Unlike the infants’, treatment can extend life but there is not cure. Respiratory
failure is the primary cause of death
C. Cori’s Disease – GSD III
a. It is caused by glycogen debranching enzyme [Amylo-1,6-glucosidase
(debrancher)] deficiency in the liver, muscles, WBC and RBC
b. AKA Forbes – Cori disease
c. In glycogenolysis, a particular enzyme is required to unlink the branch points
d. When that enzyme fails, symptoms similar to GSD I occur. In addition to GSD I
symptoms, GSD III also causes muscles prone to wasting, enlarged heart and
increased levels of lipids in the blood
D. Andersen’s Disease – GSD IV
a. It is caused by a glycogen brancher enzyme [ Alpha-1,4-glucan, 6-glucosyl-
transferase (brancher)] deficiency in the skeletal muscles
b. AKA amylopectinosis
c. The glycogen constructed in GSD IV is abnormal and insoluble
d. As it accumulates in the cells, cells die andlead to organ damage
e. Infants born with GSD IV appear normal at birth but are diagnosed with enlarged
liver and failure to thrive withn their first year.
f. Infants who survive beyond their first birthday develop cirrhosis of the liver and
by age of 3 to 5, die as a result of chronic liver failure
E. McArdle’s Disease – GSD V
a. Caused by glycogen phosphorylase deficiency in skeletal muscles (Muscle
phosphorylase)
b. Under normal circumstances, muscle cells rely on oxidation of fatty acids during
rest or light activity
c. More demanding activity requires that they draw on their glycogen store
d. In GSD V, the form of glycogenolysis is disabled and glucose is not available
e. The main symptoms are muscle weakness and cramping and a burgundy-
colored urine after exercise due to myoglobin (a breakdown product of muscle) in
the urine
F. Her’s Disease – GSD VI
a. Caused by liver phosphorylase deficiency which blocks the first step in
glycogenolysis (Hepatic phosphorylase)
b. This type of GSD is linked in the x chromosome
c. Low blood sugar is one of the key symptoms, but not as severe as the other
types of GSD’s
G. Tauri’s Disease – GSD VII
a. Caused by muscle phosphofructokinase deficiency
b. Although G6P is available as a fuel in the muscles, the cells cannot metabolize it
c. Abnormally high levels of glycogen are stockpiled in the muscle cells
d. The symptoms are similar to GSD V, but also include anemia and increased
levels of uric acid
H. Type VIII
a. Deficient enzyme: Adenyl kinase
b. Symptoms: Spasticity, decebration, high urinary cathecolamine, death in infancy
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I. Type IX
a. Deficient Enzyme: Hepatic phosphorylase b kinase
b. Symptoms: Hepatomegaly, increase liver glycogen concentrations
J. Type X
a. Deficient Enzyme: Cyclic AMP-dependent kinase
b. Symptoms: Hepatomegaly
THE PENTOSE-PHOSPHATE SHUNT

- primarily an anabolic pathway that utilizes the 6 carbons of glucose to generate 5 carbon
sugars and reducing equivalents.
- Can oxidize glucose and under certain conditions can completely oxidize glucose to
CO2 and water.
- Consists of 2 irreversible oxidative reactions followed by a series of reversible sugar-
phosphate interconversions
- No ATP is directly consumed or produced in the cycle.
- Provides a major portion of the body’s NADPH which functions as a biochemical
reductant
- Produces ribose-5-phosphate required for the biosynthesis of nucleotides
The oxidative steps

- occur at the beginning of the pathway
- the reactions that generate NADPH
- The reactions catalyzed by glucose-6-phosphate dehydrogenase and 6-
phosphogluconate dehydrogenase generate one mole of NADPH each for every mole of
glucose-6-phosphate (G6P) that enters the PPP
http://javed-abbas.tripod.com/notesonline/biochemistry/pentose-phosphate-pathway.html
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The non-oxidative reactions
- primarily designed to generate R5P
- convert dietary 5 carbon sugars into both 6 (fructose-6-phosphate) and 3
(glyceraldehyde-3-phosphate) carbon sugars which can then be utilized by the pathways
of glycolysis.
http://javed-abbas.tripod.com/notesonline/biochemistry/pentose-phosphate-pathway.html
OVERALL PURPOSE:
1. It produces 2NADPH, which is used instead of NADH in most biosynthetic pathways.

This is the perhaps the primary source of NADPH in metabolism.
2. NADPH is needed in red blood cells as a cofactor for Glutathione Reductase, which is
essential for the RBC. RBC's can utilize the pentose phosphate shunt to supply this
NADPH.
3. It produces Ribose-5-Phosphate, the essential precursor to nucleotide synthesis.
Nicotinamide adenine dinucleotide phosphate (NADP)

Produced mainly in the Pentose-phosphate pathway.
provides the reducing equivalents for biosynthetic reactions and for oxidation-reduction
involved in protection against the toxicity of ROS (Reactive Oxygen Species).
used for anabolic pathways, such as fatty acid synthesis, cholesterol synthesis and fatty
acid chain elongation.
source of reducing equivalents for cytochrome 450 hydroxylation of aromatic compounds,
steroids, alcohols, and drugs.
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PATHWAY:
1. Glucose ------> Glucose-6-Phosphate

- Catalyzed by Hexokinase; -1ATP
2. Glucose-6-Phosphate ------> 6-Phosphogluconolactone

- Catalyzed by Glucose-6-Phosphate Dehydrogenase. This is a REDOX reaction, coupled
to NADP.
- This is an internal-ester -- a lactone ring.
- Concurrent NADP ------> NADPH Reduction.
3. 6-Phosphogluconolactone ------> 6-Phosphogluconate -- a hydrolysis reaction.

- H2O is added across the lactone to open the ring.
- Catalyzed by 6-Phosphogluconolactase.
4. 6-Phosphogluconate ------> Ribulose-5-Phosphate -- a redox reaction

- Catalyzed by 6-Phosphogluconate Dehydrogenase.
- It is the oxidation of an alcohol function to a keto group, and a decarboxylation, leaving a
5-carbon-sugar skeleton.
- Concurrent reduction of NADP ------> NADPH
5. Ribulose-5-Phosphate ------> Ribose-5-Phosphate

- This is done via an isomerase.
- The reactant can also go to Xylulose-5-Phosphate via an epimerase, but that's not
metabolically important.
- The Ribose-5-Phosphate can then serve as the precursor for nucleotide synthesis.
6. Through carbon-recombination, Fructose-6-Phosphate is an ultimate product, which

can then be reconverted back to Glucose-6-Phosphate, which can then start the cycle
over again.
- Each time the pentose-phosphate pathway recycles as above, you oxidize the
equivalent of one carbon from glucose. So, if you run the cycle 6 times, you completely
oxidize one glucose.
- This oxidation of glucose is an alternative to glycolysis and the TCA cycle, which is not
energetically important, but it does provide essential intermediates -- NADPH and
Ribose-5-Phosphate.
ENTNER-DOUDOROFF PATHWAY (EDP)

- series of reactions that catabolize glucose to pyruvate.
- can occur only in prokaryotes.
- uses 6-phosphogluconate dehydrase and 2-keto-3-deoxyglucosephophate aldolase to
create pyruvates from glucose.
- has a net yield of 1 ATP for every glucose molecule processed, as well as 1 NADH and
1 NADPH.
- pseudomonads use the pathway as a way to degrade carbohydrates for respiratory
metabolism.
- The E-D pathway yields 2 pyruvic acid from glucose (same as the E-M pathway) but like
the phosphoketolase pathway, oxidation occurs before the cleavage, and the net energy
yield is one mole of ATP per mole of glucose utilized.
- In the E-D pathway, glucose phosphate is oxidized to 2-keto-3-deoxy-6-phosphogluconic
acid (KDPG) which is cleaved by KDPG aldolase to pyruvate and GAP.
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- The latter is oxidized to pyruvate by E-M enzymes wherein 2 ATP are produced by
substrate level phosphorylations.
- Pyruvic acid from either branch of the pathway is reduced to ethanol and CO 2, in the
same manner as yeast, by the "yeast-like bacterium", Zymomonas (Figure 12 below).
- Thus, the overall reaction is Glucose ---------->2 ethanol +2 CO2, and a net gain of 1
ATP.
https://www.researchgate.net/figure/Glycolysis-gluconeogenesis-pentose-phosphate-and-Entner-Doudoroff-
pathways_fig1_12823421
The Entner Doudoroff Pathway
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FORMATIVE ASSESSMENT: TBA
Scoring Rubrik for short response assignment and case analysis

4 3 2 1
unsuccessful
accuracy
correctly cited
slows readability
ASSIGNMENT:
1. What is the Luebering-Rapaport bypass? What important metabolic by-product is
produced by this pathway? What would be the effect to the RBC if there are defects in
the Luebering-Rapaport bypass?
2. In which pathway can be find glutathione? What is its importance?
3. Explain G-6-PDH deficiency. How does it affect red blood cells?
SUMMATIVE ASSESSMENT: TBA
Case:
Angel Quondensada is a 20 y/o exchange student who has noted gastrointestinal
bloating, abdominal cramps and intermittent diarrhea ever since arriving in her present country 6
months earlier. A careful history shows that these symptoms occur about 45 minutes to 1 hour
after eating breakfast but may occur after other meals as well. Dairy products, not part of
Angel’s diet from her country, were identified as the probable offending agent because her
gastrointestinal symptoms disappeared when milk and milk products were eliminated from their
diet.
Questions:
1. What is the probable condition that Angel is suffering from?
2. What is the primary cause of this condition?
3. Explain how Angel’s symptoms occurred.
4. If Angel insists on eating food with dairy, what would be your advice to her?
5. Will injury to the intestinal mucosa result to the same condition? Explain your answer
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LABORATORY
INTRODUCTION
Cellulose is a linear polysaccharide made up of beta – D glucose linked by beta – 1, 4

glycosidic bonds. These linear chains make up the fibrils which constitute the structural
component of cell walls of plants.
A fibril structure alternates between segments with a highly orderly arrangement

(crystalline segment) and randomly ordered segments (amorphous). The crystalline segment
accounts for much of the stability and strength while the amorphous region imparts flexibility
Name: _____________________________ Date: _____________________________

Activity No. 15
CELLULOSE
OBJECTIVES

1. identify correctly the basic properties of cellulose
2. differentiate accurately the properties of cellulose from starch and glycogen
3, perform precisely simple tests to determine the properties of cellulose
Scoring rubric
For Procedures:
For Rationalization
not clear
not clear
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Solubility
1. Get 4 test tubes and label them with the solvent to be used: Water, dilute HCl, conc. HCl
and conc. NaOH
2. Get a cotton ball and get a few fiber strands and place them in the labelled test tubes
3. Add 2 mL of the corresponding solvents
4. Try to the dissolve the cotton fibers by shaking the test tube carefully from time to time.
5. Repeat the procedure but his time, use the fibers coming from filter papers
6. Write your observations in the table below
Observations:
Solvents Used Cotton Balls

Water, H2O
Dil. HCl
Conc. HCl
Conc. NaOH
What conclusion can you derive from these observations?

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
Iodine test
1. Get a cleat test tube and place a few cotton strands
2. Add a few deops of Lugol’s iodine
3. Result
___________________________________________________________________
What is your interpretation of this result?
___________________________________________________________________
How does cellulose differ from other polysaccharides in terms of its reaction to iodine?
(recall how other polysaccharides react to iodine)
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
Scoring Rubric
1. Discuss how cellulose can be hydrolyzed.
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2. As seen in the activity, cellulose is almost insoluble in different types of solvents, what
solvents can possibly dissolve it? Name its composition.
3. Can humans metabolize cellulose? What is the importance of cellulose in the body?
4. Differentiate starch, glycogen and cellulose as to their monosaccharide unit,
disaccharide unit, main mode of bonding and occurrence.
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Week no. 11
Submodule no. 11
Amino acids and

polypeptides
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LESSON: Amino Acids and Polypeptides
TOPICS:
7.1. Amino Acids
7.1.1. Definition and General Structure
7.1.2. Properties and Importance
7.1.3. Classification
7.1.3.1. According to the Variable group
7.1.3.2. According to Dietary Need
7.2. Polypeptides
TIME /SCHEDULE: Week no. 13
LECTURE: 5 hours
LABORATORY: 4 hours
LEANING OBJECTIVES:
At the end of the learning session, the student should be able to:
1. Define correctly the different terms encountered in this unit;
2. Classify correctly proteins according to structure and function;
3. Illustrate accurately the structures of the different amino acids;
4. Contrast accurately the different categories of protein structure;
5.Illustrate accurately the digestion and absorption of dietary proteins
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BCHMLS 1
LESSON PROPER
INTRODUCTION
One of the biomolecules discussed in the introduction is protein. An extraordinary number

of different proteins, each with a different function, exist in the human body. A typical human cell
contains 9000 different kinds of proteins, and the human body contains 100,000 different proteins.
Proteins are needed for the synthesis of enzymes, certain hormones, and some blood
components; for the maintenance and repair of existing tissues; for the synthesis of new tissues;
and sometimes for energy. (Stoker, 2011)
Lesson 1. Definition of Proteins, Structure and Classification of Proteins
Activity 1.
1. Given the following words, come up with your own definition of protein.
Organic, amino acids, carbon, body-building, macromolecule, hydrogen, oxygen, tissue
repair, nitrogen, large amount, building blocks
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________.
2. Illustrate the structure of amino acid and label each component.
Proteins are organic compounds of high molecular weight made up of alpha-amino acids
joined by means of peptide linkage. They are the most important of all biological substances,
being the fundamental constituent of the protoplasm of the cells. Proteins was derived from the
Greek word “proteios” which means “of first importance” or “to take first place.”
Proteins supply the body not only with heat and energy, but also provide materials for
building and repair. They are the “working molecules” which perform operational functions in living
systems (e.g. enzymes, antibodies, hormones, etc.) and composed of amino acids as building
blocks.
Amino Acids: The Building Blocks of Proteins

Amino acid is an organic compound that contains both an amino (-NH2) group and a
carboxyl (-COOH) group. AA in proteins are always the α-amino acids. The different components
of the amino acid structure have corresponding functions.
C (alpha carbon) is tetrahedral, chiral, asymmetric and covalently linked to both an amino
group and a carboxyl side chain. Bonded to it is a H atom and a variable R group.
R side chain varies and give the amino acid its characteristics. The nature of this side
chain distinguishes a-amino acids from each other. Side chains vary in size, shape, charge,
acidity, functional groups present, hydrogen-bonding ability, and chemical reactivity.
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BCHMLS 1
NH2 or amino group exists as –NH3+ in neutral solutions.
COOH or the carboxyl group exist as –COO- in neutral solution.
CLASSIFICATION OF AMINO ACIDS

1. Non-polar amino acids
• Hydrophobic amino acids. When incorporated into a protein, they are not attracted to water
molecules. They are generally found in the interior of proteins, where there is limited
contact with water.
• Include all those with alkyl side chain R groups
o Alanine
o Valine
o Leucine
o Isoleucine
• Cyclic structure
o proline
• sulfur containing amino acids
o cysteine and methionine
• Aromatic amino acids
o phenylalanine and tryptophan
2. Polar, uncharged amino acids

• Can form hydrogen bonds with water except glycine
• Usually more soluble in water than the non-polar amino acids
• Good hydrogen-bond forming moieties
o The amide groups of asparagine and glutamine
o The hydroxyl groups of tyrosine, threonine and serine
o Sulfhydryl group of cysteine
• Glycine – simplest amino acid
o Has only a single hydrogen for an R group and is not a good hydrogen bond former
3. Acidic amino acids
• R groups contain a carboxyl group
o Aspartic acid
o Glutamic acid
• They have net negative charge at pH 7
• Play an important role in protein functions
• Proteins that bond metal ions for structural or functional purposes possess metal binding
sites containing one or more aspartate and glutamate side chains
4. Basic amino acids

• With net positive charge at neutral pH
o Histidine
o Arginine
o lysine
• Histidine side chains play important roles as proton donors and acceptors in many enzyme
reactions
• Histidine containing peptides are important biological buffers
5. *other amino acids

• Occur only rarely in proteins
• Found mainly in the collagen and gelatin proteins
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BCHMLS 1
o Hydroxylysine
o hydroxyproline
• Found only in a light-driven proton-pumping protein call bacteriorhodopsin
o Pyroglutamic acid
TYPES OF AMINO ACIDS

1. Alpha amino acids – the amino acid found in proteins where the amino group is attached to
the 1st carbon atom adjacent to the carboxyl group
2. Beta amino acids – this type of amino acids is names as such because the amino group is
attached to the 2nd carbon atom adjacent to the carboxyl group
3. Gamma amino acids – the amino acid where the amino group is attached to the 3 rd carbon
atom adjacent to the carboxyl group
3 GROUPS OF AMINO ACIDS

1. Neutral A.A. – when their molecules have the same number of amino and carboxyl groups
• Alanine (Ala) – non polar
• Aspargine (Asp) – polar
• Cysteine (Cys) – polar
• Glutamine (Gln) – polar
• Glycine (Gly) – non-polar
• Isoleucine* (Ile) – non-polar
• Leucine* (Leu) – non-polar
• Methionine* (Met) – non-polar
• Phenylalanine* (Phe) – non-polar
• Proline (Pro) – non-polar
• Serine (Ser) – polar
• Threonine* (Thr) – polar
• Tryptophan* (Trp) – non-polar
• Tyrosine (Tyr) – polar
• Valine* (Val) – non-polar
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BCHMLS 1
2. Acidic A.A. – when their molecules have more carboxyl groups than amino groups
• Aspartic acid (Asp)
• Glutamic acid (Glu)
3. Basic A.A. – when their molecules have more amino groups than carboxyl groups
• Lysine* (Lys)
• Arginine (Arg)
• Histidine (His)
*Essential amino acids
ESSENTIAL VS NON-ESSENTIAL
Essential Amino Acids – are those amino acids that cannot be synthesized by the body and are required in the
diet. Since the body is not capable of synthesizing them, they must be supplied in our diet if we are to enjoy
normal health
Non-essential amino acids – are those amino acids that can be synthesized by the body and need not be
included in the diet
Activity 2. List the 20 amino acids and write their three letter and one letter abbreviation.
Lesson 2. Chemical Properties of Amino Acids
Previously, the definition of protein and amino acids, and their classifications was discussed. This lesson
will talk about the different chemical properties of amino acids? Recall your discussion of inorganic chemistry.
Can you remember examples of chemical changes and properties? A good memory on that particular topic will
help you understand the different tests for amino acids.
Reactions are dependent on the numerous different reactive groups that are present in the same
molecule. Aliphatic monoamino monocarboxylic acids give all the reactions expected for carboxyl and amino
groups. In addition, some reactions are characteristic of additional groups that may be present. For instance,
cysteine give the reactions characteristic of the sulfhydryl
(-SH) group and tyrosine give the reaction characteristic of phenolic group. These reactions may also be
exhibited by the protein containing these amino acids.
Ninhydrin test
• General test for amino acids
• Reagent : triketohydrindene hydrate
• Product: CO2, ammonia and an aldehyde containing one less carbon than the amino acid
• Positive result: blue or purple color
o proline & hydroxyproline gives yellow color
o Color is produced by the ammonia group
• Alpha amino acid + Ninhydrin = diketohydrindylidene-diketohydrindamine + CO2 + aldehyde
Reactions of specific amino acids
1. Millon Reaction
• reagent: Millon’s reagent; Hg(NO3)2 in HNO3 w/ trace HNO2
• Positive result: red color
• Specific for Tyrosine
2. Sakaguchi test
• reagent: α-naphthol and sodium hypochlorite
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• Positive result: red color
• Specific for: guanidine and arginine
3. Nitroprusside test
• Reagent: sodium nitroprusside [Na2(NO)5.2H2O] in dilute ammoniacal solution
• result: red color
• Specific for cysteine and proteins with free sulfhydryl group
4. Aldehyde reaction
• Indole derivative gives strongly colored products with a number of aromatic aldehydes. With
paradimethylaminobenzaldehyde in H2SO4, a red violet color is obtained with tryptophan (Ehrlich
reaction). This test can be used for the quantitative estimation of tryptophan in proteins
5. Folin reaction
• In alkaline solution, amino acids give a deep red color with sodium 1,2-naphthoquinone-4-sulfonate
• Used for rapid quantitative estimation of amino acids
6. Pauly reaction for Histidine and Tyrosine

• Histidine and tyrosine couple with diazotized sulfanilic acid in alkaline solution giving a red color
Lesson 3. Peptides
A peptide is an unbranched chain of amino acids, each joined to the next by a peptide bond. Since amino
acids contain both amino and carbonyl groups, they combine with each other to form amides, called peptides.
The linkage that joins them is known as a peptide linkage.
The product is an amide made up of two amino acids joined together, called a dipeptide
CLASSIFICATION OF PEPTIDES
• If three amino acid units are included in a molecule, it is a tripeptide, if 4 a tetrapeptide, if 5 a pentapeptide
and so on
• Oligopeptides – peptide chain of more than 12 residues and less than about 20
• Polypeptides are peptides containing about 40 – 50 amino acid units in a chain
• Amino acid residues – units making up amino acids minus the elements of water
• Proteins – larger chains of amino acids
peptides
By convention, peptide formation is written with the free amino group on the left and the COOH on the
right. In linear peptides, one end of the chain has a free amino and the other end a free carboxyl group. The
amino group end is called the N terminal residue and the other end the C terminal residue
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1 2 3 4 5 6 7 The sequence of amino acids in a chain is
Ala – pro – tyr – met – gly – lys – gly numbered starting with the N-terminal
residue, which is written on the left, the C-
N-terminal residue C-terminal residue terminal residue is written on the right
peptides Any segment of the sequence that is not
specifically known is placed in parenthesis
NOMENCLATURE OF PEPTIDES
• Named as acyl derivatives of the C-terminal amino acid, the C-terminal unit keeping its complete name
• The –ine ending of all but the C-terminal amino acid is changed to –yl
• Except tryptophan (tryptophyl), cysteine (cysteinyl), glutamine (glutaminyl), asparagine (asparaginyl)
• They are listed in the order in which they appear, starting with the N-terminal amino acid
Example: Ala-Tyr-Gly is called alanyltyrosylglycine
Arg-Glu-His-ala is arginylglutamylhistydylalanine
Activity 3. PRACTICE NAMING. Identify the number of amino acids, number of peptide bonds, and name the
peptide.
i.e. Ala-Cys
- dipeptide
- 1 peptide bond
- Alanylcysteine
1. Arg-pro-pro-gly-phe-ser-pro-phe
________________________________________________________________________________________
________________________________________________________________________________________
____________________________________________________
2. Cys-tyr-phe-gln-asn-cys-pro-arg-gly
________________________________________________________________________________________
________________________________________________________________________________________
____________________________________________________
3. Glu-leu-tyr-glu-asn-lys-pro-arg-arg-pro-tyr-ile-leu
________________________________________________________________________________________
________________________________________________________________________________________
____________________________________________________
4. Tyr-gly-gly-phe-met
________________________________________________________________________________________
________________________________________________________________________________________
____________________________________________________
Lesson 4. Polypeptides
The amino acid sequence and chain length give a polypeptide its biological effectiveness. The sequence
of amino acid residues is essential for proper polypeptide function. This sequence aligns the side-chain
characteristics in the proper positions for a specific polypeptide function.
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PRIMARY STRUCTURES AND FUNCTIONS OF SOME BIOLOGICAL POLYPEPTIDES
Name Primary Structure General Biological Functions
Substance F Arg-Pro-Lys-pro-Gln-Gln-Phe- A pain producing agent

Phe-gly-leu-met-NH2
Bradykinin Arg-pro-pro-gly-phe-ser-pro-phe Affects tissue inflammation and blood pressure
Angiotensin II Asp-arg-val-tyr-val-hist-pro-phe Maintains water balance and blood pressure
Leu-enkephalin Tyr-gly-gly-phe-leu Relieves pain, produces sense of well being

Met-enkephalin Tyr-gly-gly-phe-met
Vasopressin Cys-tyr-phe-gln-asn-cys-pro-arg- Increases blood pressure, decreases kidney
gly-NH2 water excretion
Oxytocin Cys-tyr-ile-gln-asn-cys-pro-arg- Initiates childbirth labor, causes mammary gland
gly-NH2 milk release, affects kidney excretion of water
and sodium
Thyrotopin- Glu-His-Pro Stimulates release of hormones from the pituitary
releasing gland
hormone
Orexin 33 amino acids long Causes wakefulness
Neuropeptide Phe-leu-phe-gln-pro-gln-arg-phe- Modulates pain sensation

FF NH2
Neurotensin Glu-leu-tyr-glu-asn-lys-pro-arg- Involved in brain memory functions
arg-pro-tyr-ile-leu
Neuropeptide Y 36 amino acids long Stimulates eating
Endomorphin Tyr-pro-trp-phe-NH2 Acts as a morphine-like analgesic
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LABORATORY
INTRODUCTION
Proteins have many functions in the body. They are macromolecules consisting of long sequences of
alpha amino acid in peptide linkages. They are considered as working molecules and the building blocks of our
body. Thus virtually almost all tissues in the body contain proteins. For today’s activity, you will perform two
tests that detect the presence of proteins in a sample.
Name: ___________________________________ Date: _____________________________

Date Assigned: ____________________________ Date submitted: _____________________
Activity 16
GENERAL TESTS FOR PROTEINS

OBJECTIVES

1. discern properly the principle behind the Biuret and Ninhydrin test;
2. recognize correctly the limitations of these tests.
MATERIALS
Reference material pen/pencil

Scoring rubric
For Procedures:
o 2 points for correct photo downloaded with label
- 1 point is given for items that has observation but without downloaded photo
not clear
not clear
Ninhydrin test
1. Should you have six test tubes with the following test materials: egg albumin, milk, tyrosine, tryptophan,
methionine and arginine and then added with Ninhydrin reagent and heated in a water bath for 2 ½
minutes, what would the possible result be? Write it in the observation below and download a photo of
the positive result for each sample. Attach the photo to your paper.
Test Observation Interpretation
Egg albumin
Milk
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Tyrosine
Tryptophan
Methionine
Arginine
- Which of the substances gave a positive result to ninhydrin test?

___________________________________________________________________
- Which of the substances tested are proteins?
___________________________________________________________________
- Which of the substances tested are amino acids?
___________________________________________________________________
- What is the Ninhydrin test for?
___________________________________________________________________
- Why did all substances above gave positive results to Ninhydrin test if this test is considered a general
test for amino acids?
__________________________________________________________________________________
__________________________________________________________________________________
_____________________________________
- Will all amino acids give the same color result to Ninhydrin test?
___________________________________________________________________
- If not, which amino acids will give different colored results to Ninhydrin test?
__________________________________________________________________________________
____________________________________________________
Biuret test
1. Should you have 5 test tubes with the following samples: egg albumin, milk, tyrosine, arginine and
tryptophan. To each of these tubes, you added 10% NaOH solution and dilute CuSO4 solution while
swirling, what will the possible result be? Write your answer on the data sheet provided below.
Download a photo of the positive result and attach it to your paper.
Observations
Substance tested Result Interpretation
Egg albumin
Milk
Tyrosine
Tryptophan
➢ Will all substances give a positive result?
__________________________________________________________________________________
➢ If not, why?
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________

Scoring Rubric
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1. What is the principle behind

a. Biuret test
b. Ninhydrin test
2. State the limitation(s) of the Biuret test and the Ninhydrin test
3. Which is a better test for proteins, Biuret or Ninhydrin? Defend your answer.
4. Enumerate and describe the different functions of proteins.
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INTRODUCTION
Proteins are very important biological molecules with many functions. Functions of proteins would include
transport of substances, storage, movement, structure and catalysis. Proteins are considered macromolecules
and are polymers of amino acids – that is they consist of long chains of amino acids held by peptide bonds
Amino acids are the building blocks of proteins. It contains an amino group and a carboxyl group and a side
chain that varies from one amino acid to the other and this results to 20 naturally occurring amino acids. The
reactions of amino acids is associated with the different functional groups found in its structure.
NAME: _____________________________ DATE: _______________________________

DATE ASSIGNED: _____________________ DATE SUBMITTED: _____________________
Activity 17
SPECIFIC TESTS FOR AMINO ACIDS
OBJECTIVES
1. identify correctly the different specific tests for amino acids
2. state accurately the principle behind each of the tests
MATERIALS
Scoring rubric
For Procedures:
o 1 point for the answer/observation
For Rationalization
not clear
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not clear
Millon’s test
1. Should you have two test tubes with milk and egg albumin and you added each with Millon’s reagent
and heated each in the water bath for 3 minutes, what would be the possible result? Write your answer
on the data table. Download a photo of the positive result and attach it to your paper.
2. What amino acid/s is indicated by a positive Millon’s test?
___________________________________________________________________
Xanthoproteic test
1. Should you have two test tubes with milk and egg albumin and you added each with concentrated nitric
acid and heated each in a warm water bath for 3 minutes. After which you added conc. NaOH until
alkaline, what would be the possible result? Write your answer on the data table. Download a photo of
the positive result and attach it to your paper.
2. What amino acid/s is indicated by a positive Xanthoproteic test?
__________________________________________________________________________________
____________________________________________________
Hopkin’s-cole test
1. Should you have two test tubes with milk and egg albumin and you added each with Hopkin’s Cole
reagent and then you added 30 drops of conc. Sulfuric acid along the side of the test tube, what would
be the possible result? Write your answer on the data table. Download a photo of the positive result and
attach it to your paper.
2. What amino acid/s is indicated by a positive Hopkin’s-Cole test?
__________________________________________________________________________________
__________________________________________________________________________________
Lead Acetate test
1. Should you have two test tubes with milk and egg albumin and you added each with 10% NaOH
solution and lead acetate solution, and heated each in a boiling water bath for 5 minutes then cool,
what would be the possible result? Write your answer on the data table. Download a photo of the
2. What amino acid/s is indicated by a positive lead acetate test?
__________________________________________________________________________________
__________________________________________________________________________________
Sakaguchi test
1. Should you have two test tubes with milk and egg albumin and you added each with 10% NaOH and
dilute alcoholic a-naphthol solution, mixed then added with sodium hypochlorite solution, what would be
the possible result? Write your answer on the data table. Download a photo of the positive result and
2. What amino acid/s is indicated by a positive Sakaguchi test?
__________________________________________________________________________________
__________________________________________________________________________________
Sodium nitroprusside test

1. Should you have two test tubes with milk and egg albumin and you added each with 1% sodium
nitroprusside and 10% NaOH, what would be the possible result? Write your answer on the data table.
Download a photo of the positive result and attach it to your paper.
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2. What amino acid/s is indicated by a positive sodium nitroprusside test?
__________________________________________________________________________________
__________________________________________________________________________________
Data Table
Test Milk Albumin conclusion

Scoring Rubric
1. Tabulate the different tests for proteins performed in this activity PLUS the ninhydrin and Biuret test
showing the following: test, reagent, positive result, structure in the protein/amino acid tested and the
specific amino acid tested for.
2. State the principle involved in each test performed
3. What makes glycine, cysteine, tyrosine and proline different from other amino acids?
4. Why can’t nitric acid stains be removed by washing?
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WEEK NO. 12
SUBMODULE NO. 12
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PROPERTIES OF
PROTEINS
LESSON: PROPERTIES OF PROTEINS
TOPICS
7.2. Definition and Functions of Proteins
7.3. Classification of Proteins
7.4. Categories of Protein Structure
7.4.1. Primary Structure
7.4.2. Secondary Structure
7.4.3. Tertiary Structure
7.4.4. Quarternary Structure
7.5. Digestion and Absorption of Dietary Proteins
TIME/SCHEDULE: WEEK 14
LECTURE: 5 HOURS
LABORATORY: 4 HOURS
LEARNING OBJECTIVES
1. Define correctly the terms related to this unit
2. Classify proteins correctly
3. Contrast accurately the different categories of protein structure
4. Illustrate accurately the digestion and absorption of dietary proteins
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LESSON PROPER
INTRODUCTION:
PROTEINS
Structure of proteins
• Proteins contain the elements C, H, O, N and usually S (and P) plus traces of Fe, Cu, I, Mn and Zn
• Amino acids – building blocks of proteins
3 Categories of Protein Structure

Primary Structure
• The sequence of amino acids which are joined together to form the polypeptide
• The main mode of linkage is the peptide bond
• Primary structure is the order of attachment of the AA to each other through peptide bond. It is always
the same regardless of where protein is found in an organism. i.e. insulin
• It is the most important because it is the protein’s AA sequence that determines its overall shape, function
and properties.
• Crucial that a change of only one AA can drastically alter its biological function.
o i.e. Sickle-cell anemia val is substituted by glu
Secondary Structure
• The peptide chains are folded regularly, the folding resulting from the linkage of the carbonyl group of
one peptide chain with the amine group of another chain by means of hydrogen bonds
• This hydrogen bonding produces a regular coiled arrangement called Helix
• α-helix found in α-keratins that make up the hair, skin and nails of humans
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• Secondary structure result primarily from hydrogen-bonding interactions b/n different amide linkages
along the protein backbone.
Tertiary structure
• The tertiary structure of the protein arises from the interactions that take place among the side groups of
polypeptide chains, interactions which cause bending and folding of the protein molecule
• Maintained by covalent disulfide bonds
• Each type of protein molecule has in its native state, a characteristic 3-dimensional shape referred to as
conformation (refer to the combined 2o and 3o structure of the peptide chain
• Tertiary forms the overall 3-dimensional shape of a protein that results from the interaction between AA
side chains
2 major groups of proteins

• Globular proteins – named to describe the spherical shape into which their polypeptide chains are bent
and folded as a result of tertiary structure
o Perform the operational functions in the body by behaving as enzymes, hormones, antibodies
and transporters (Hgb and serum albumin)
• Fibrous proteins – made up of polypeptide chains arranged in parallel fashion along a single axis, to
yield long fibers or sheets
• Basic structural unit of connective tissues
• Ex. Collagen of tendons and bone matrix, α-keratin of hair, horn, nails, Elastin (elastic connective
tissue)
• Some proteins (e.g. myosin and fibrinogen) are intermediate between fibrous and the globular, Rod-
like structures (like the fibrous) and like the globular, they are soluble in aqueous salt solutions
An analogy for the relationships among the primary, secondary, and tertiary structures of a protein is that
of a telephone cord. The primary structure is the long straight cord. The coiling of the cord into a helical
arrangement gives the secondary structure. The supercoiling arrangement the cord adopts after you hang the
receiver is the tertiary structure. (Stoker, 2011)
Tertiary
Structure
Primary
Structure
Secondary
Structure
Quaternary/quaternary protein structures

• Protein organization is produced by fitting together separate coiled and folded structures to form an
aggregate functional structure
• Ex. Isoenzymes (enzymes with similar but not identical catalytic powers)- LDH, hexokinase,
phosphatases
• Oligomeric protein – proteins with more than one chain
o Protomers – component chain
o Ex. Hemoglobin – made up of 4 polypeptide chain (2α and 2β), it contains 140 amino acids in
each chain
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Lesson 5. Proteins and Their Biochemical Functions
Activity 4. Translate the functions of proteins into drawings. Make sure to label your drawing properly. The
objective of this activity is to determine your understanding of the topic. (20 points)
CRITERIA 4 (Excellent) 3 (Good) 2 (Fair) 1 (Poor)

Content/ The student’s The student’s The student’s The student’s
Interpretation X 2.5 illustration depicts a illustration shows a illustration depicts a illustration shows a
very clear picture of clear picture of the somewhat clear different interpretation
the functions of functions of proteins. picture of the different on the different
proteins. All Almost all functions of protein. functions of protein.
components of the components of the Some components of The components
illustration show the illustration show the the illustration show included are not
overall idea of the overall idea of the the idea of the significant to support
student. Further, the student. Further, the student but lacks the student’s idea of
drawing is very easy drawing is easy to some details. Further, the identified function.
to understand and understand and not all drawing is Further, the drawing
interpret. interpret should a easy to understand is vague to
brief discussion is and interpret. understand.
included.
Graphics – Several of the One or two of the The graphics are No graphics made by
Originality graphics used on the graphics used on the made by the student, the student are
X 1.25 illustration reflect an illustration reflect but are based on the included
exceptional degree of student creativity in designs or ideas of
student creativity in their creation and/or others.
their creation and/or display.
display.
Attractiveness and The poster is The poster is The poster is The poster is
Organization X 1.25 exceptionally attractive in terms of acceptably attractive distractingly messy or
attractive in terms of design, layout and though it may be a bit very poorly designed.
design, layout, and neatness. Some messy. Not all It is not attractive. It is
neatness. It is also drawings are not drawings were not labeled as well.
labeled properly. labeled. labeled accordingly.
Proteins and their Biochemical Functions
1.Structure– for animals, it is structural proteins which are the chief constituents of skin, bones, hair and
fingernails
• Collagen – found in bones – provides the support for calcium to bind and form the solid structure of the
bones
• Keratin – hair and nails
2. Catalysis– virtually all the reactions that take place in the living organisms are catalyzed by proteins called
enzymes. The enzyme trypsin – catalyzes hydrolysis of proteins; sucrose – catalyzes the hydrolysis of sucrose;
and dehydrogenase – converts ethanol to acetaldehyde. Without enzymes, the reactions would take place so
slowly as to be useless.
3. Movement and Contraction– every time we crook our finger, climb stairs, or blink an eye, we use our
muscles. Muscles expansion and contraction are involved in every movement we make. Muscles are made up
of protein molecules called myosin and actin
4. Elasticity– the protein elastin, which possesses unique elasticity and strength, enables the skin, ligament and
blood vessels to stretch and rebound
5. Transport– a large number of proteins fall into this category

• Albumin – transports bilirubin, calcium, fatty acids, some drugs
• Ferritin and transferrin – transport iron
• Transcortin – cortisol (cortisol-binding protein)
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• Hemoglobin – carries oxygen
• Lipoproteins – cholesterol and triglycerides
6. Hormones– Many hormones are proteins, among them insulin, oxytocin, human growth hormone and
endomorphines.
7. Protection– when a protein from an outside source or other substance (called antigen) enters the body, the
body makes its own proteins (called antibodies) to counteract the foreign protein. This is the major mechanism
the body used to fight diseases. Blood clotting is another protective device carried out by proteins called
fibrinogen. Without clotting, we would bleed to death from any small wound
8. Storage– some proteins are used to store materials, in the way that starch and glycogen store energy.
Examples are casein in milk and ovalbumin in eggs, which store nutrients for newborn mammals and birds.
Ferritin, a protein in the liver, stores iron
9. Regulation – some proteins control the expression of genes are thereby regulate the kind of proteins
manufactured in a particular cell, and control when such manufacture takes place
Classification of Proteins on a Nutritional Basis

Complete proteins
• Proteins that supply all the essential amino acids needed by the human body for normal health
• They are derived from animal sources
• They are also referred to as high biological value protein.
Incomplete proteins
• Are the proteins that are deficient in one or more essential amino acids
• Many proteins especially those from vegetable sources, are incomplete thus the other name which low
biological value protein.
• For example, protein from corn (maize) is deficient in lysine
TWO MAJOR GROUPS OF PROTEIN:

SOME COMMON FIBROUS AND GLOBULAR PROTEINS
NAME OCCURRENCE AND FUNCTION
Fibrous Proteins (Insoluble)
Keratins Found in wool, feathers, hooves, silk, and fingernails
Collagens Found in tendons, bone, and other connective tissues
Elastins Found in blood vessels and ligaments
Myosins Found in muscle tissue
Fibrin Found in blood clots
Globular Proteins (Soluble)
Insulin Regulatory hormone for controlling glucose metabolism
Myoglobin Involved in oxygen storage in muscles
Hemoglobin Involved in oxygen transport in blood
Transferrin Involved in iron transport in blood
Immunoglobulins Involved in immune system responses
Source: Biochemistry, Stoker, 2011.
Lesson 6. Classification of Proteins
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CLASSIFICATION OF PROTEINS
I. SIMPLE PROTEINS- true proteins found abundantly in both plants and animals. On hydrolysis with enzymes,
they yield α-amino acids and their derivatives
a. Albumins- soluble in water & dil. Neutral salt solution
- coagulated by heat & precipitated by full saturation with (NH4)2SO4 but not with NaCl except in the presence of
acid
- Members: serum albumin – blood
Lactalbumin- milk
Ovalbumin- egg white
b. Globulins- soluble in neutral dil. Salt solution but NOT water

-coagulated by heat and can be precipitated from their solutions by half saturation with (NH 4)2SO4 and complete
saturation with NaCl
-Members: Ovoglobulin- egg white
Edestin- hempseed
Legumin- peas
Myosinogen- muscles
Serum globulin
c. Glutelins- soluble in dil. Acids and alkalies but insoluble in neutral solvents
- Glutenin- wheat
- Oryzenin- rice
d. Prolamines- insoluble in ordinary solvent but soluble in 70% alcohol

- not coagulable by heat
- present in plants (e.g. Gliadin- wheat, Zein- corn, Hordein- barley)
e. Histones- soluble in water, dilute acids & alkalies but not in dil. Ammonia
- not readily coagulated by heat
- strongly basic and occur in tissues in the form of salt - combinations with acid substances like heme of the
hemoglobin molecule
Globin- hemoglobin
Histone- thymus
Scombrone- mackerel
f. Protamines- contain small number of amino acids

- soluble in water & dilute acids and alkalies and are not coagulated by heat
- strongly basic; form soluble salts with strong mineral acids
Salmin- salmon sperm
* Prolamine vs. prolamine – protamine has low MW that are rich in arginine and are found associated especially
with DNA in place of histone in the sperm cells of various animals (as fish)
g.Scleroproteins (Albuminoids)- insoluble in water and neutral solvents

Keratin- epidermal tissues
Elastin- ligaments
Collagen- hides, bones & cartilages
II. CONJUGATED PROTEINS- made up of protein molecules combined with non-protein groups
a. Nucleoproteins- combinations of histones and protamines with nucleic acid
- soluble in dilute solutions of NaCl and can be extracted from the tissues by the use of this solvent; precipitated
by acidification
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Chromatin
Products obtained from glandular tissues
Germ of grains
b. Glycoproteins- proteins with a carbohydrate component

utilized for lubricating purposes in view of their slimy nature
Mucin- saliva
Tendomucoid- tendons
Osseomucoid- bones
- Glycoproteins are not digested by the enzymes of the gastro-intestinal tract, thus help in protecting the
membranes of the tract against digestion.
c. Phosphoproteins- prosthetic group (H3PO4) joined to the protein molecule

Casein- milk
Vitellin- egg yolk
d. Chromoproteins- protein compounds with hematin or similar pigments

Hemoglobin, cytochromes, rhodopsin
e. Lipoproteins- fatty substances combined with their molecules like lecithin, cephalin, etc.
blood serum, brain tissues, cell nuclei,
egg yolk and milk
TYPES OF CONJUGATED PROTEINS

Class Prosthetic Group Specific Example Function of Example
Hemoproteins Heme unit Hemoglobin Carrier of O2 in blood
myoglobin Oxygen binder in muscles
Lipoproteins Lipid Low-density lipoprotein Lipid carrier
(LDL)
High-density lipoprotein Lipid carrier
(HDL)
Glycoproteins Carbohydrate Gamma globulin Antibody
Mucin Lubricant in mucous secretion
Antiviral protection
Interferon
Phosphoproteins Phosphate group Glycogen phosphorylase Enzyme in glycogen phosphorylation
Nucleoproteins Nucleic acid Ribosomes Site for protein synthesis in cells
Self-replicating, infectious complex
viruses
Metalloproteins Metal ion Iron-ferritin Storage complex for iron
Zinc-alcohol DH Enzyme in alcohol oxidation
Source: Biochemistry, Stoker, 2011.
III. DERIVED PROTEINS- substances formed from simple and conjugated proteins
A. Primary Protein derivatives- synonymous with denatured proteins; have undergone slight intramolecular
rearrangement through the hydrolytic action of certain physical and chemical agents
• Proteans- insoluble substances resulting from the preliminary action of water, dilute acids or enzymes
Ex. Myosan (from Myosin)
Edestan (from Edestin)
• Metaproteans- products of further hydrolysis; soluble in weak acids and alkalies

Ex. Acid metaproteans (acid albuminate)
Alkali metaproteans (alkali albuminate)
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• Coagulated proteins- insoluble products resulting from either the action of heat, alcohol, UVR or even
simple mechanical shaking
Ex. Cooked egg albumin, cooked meat
B. Secondary Protein derivatives- products of more extensive hydrolysis

mixtures of fragments of original protein varying in composition and size
- exhibit common properties (e.g. solubility in water and non-coagulability by heat)
1. Primary proteoses- soluble in water, precipitated by conc. HNO3 & half saturation with (NH4)2SO4 or ZnSO4;
not coagulated by heat
2. Secondary proteoses- precipitated by complete saturation with (NH4)2SO4
3. Peptones- soluble in water; precipitated by saturation with certain alkaloidal reagents (e.g. Phosphotungstic
acid, Tannic acid)
4. Peptides- combinations of 2 or more amino acids, the carboxyl group of one being united with the amino
group of the other
Lesson 7. Physical and Chemical Properties of Protein
PHYSICAL AND CHEMICAL PROPERTIES OF PROTEINS

The number, kind and arrangement of the various amino acids within the protein molecule are responsible
for the variety of complex proteins. However, protein substances share certain COMMON properties which serve
to identify them as a distinct class of biological substances.
- when pure, proteins are generally tasteless
- colorless, amorphous compounds
- insoluble in fat solvents & present varied degrees of solubility in water, salt solution, dilute acids and
alkalies
- due to high MW, form non-diffusible colloid solutions of the emulsoid type
- proteins are amphoteric; they are labile, readily modified in solution when subjected to alterations in pH,
UVR, heat and organic solvents
- very reactive & highly specific due to the presence of side chains & active NH 2 groups
SOLUBILITY
Various kinds of proteins differ markedly in their solubility; utilized for group separation and most often
for purification of individual proteins.
Factors Affecting Solubility:

1. Neutral salt- low concentration of neutral salts increases the solubility of proteins (“salting in” effect) while an
increase in concentration decreases solubility (“salting out” effect).
The solubility of any substance depends upon the relative affinity of solute molecules for each other and
for the solvent. Any factor which decreases the interaction of solute molecules will tend to increase solubility.
2. Effect of pH- solubility is influenced by pH because of their amphoteric nature; solubility is minimum at
isoelectric point and increases with increasing acidity or alkalinity
3. Effect of Organic solvents

When an organic solvent (which is miscible in water—e.g. methanol, ethanol, acetone) is added to
aqueous solution of proteins, it lowers the dielectric constant, so that the electrical forces between the charged
particles in the solution are increased, thus diminishing the solubility of proteins.
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Selective separation of the constituent proteins of a mixture can be achieved by proper adjustment of pH,
temperature, alcohol concentration, protein concentration and salt concentration kept at low level.
E.g. Fractionation of plasma with this procedure produces fractions which are of clinical significance:
Fibrin film & foam – used for surgery
Albumin – treatment of shock, nephrosis, cirrhosis
Globulin – passive immunization
Agglutinins – for blood typing
ACTION OF HEAT
- when burned, proteins decompose and liberate a characteristic odor of burned hair or feather
Denaturation- the process by which solution of proteins undergo slight intramolecular rearrangements when
they are heated between 38 to 60 degrees Celsius, giving rise to changes in chemical, physical and biologic
properties.
• Believed to be due to the unfolding of the characteristic folded structure of polypeptide chain.
• Reversible process; the unfolded molecule will return to its native form when conditions become favorable
(Renaturation, Refolding or Annealing)
• Renders the protein insoluble at isoelectric point, causing it to precipitate (Flocculation)
Coagulation- If flocculated protein is heated further, the clumped chains become matted together in a mass
which is insoluble, not only at isoelectric point but also over the entire pH range
• a process involving the linkage of adjacent protein molecules by means of side chain Hydrogen bonds
• coagulated protein is hard to dissolve & when dissolved, it differs from the original protein from which it
is derived; process is irreversible.
Other Factors which bring about Denaturation:

1. Low temperature
2. high pressure
3. UVR – operates very similarly to the action of heat (i.e. sunburning)
4. surface action
5. mechanical shaking or violent whipping – causes molecules in globular shapes to extend to longer
lengths, which then entangle (i.e. beating egg with white into meringue)
6. chemical agents (e.g. acids, alkalies, organic solvents, alcohol, etc.).
a. strong acids and bases disrupt hydrogen bonds and salt bridges; prolonged action leads to actual
hydrolysis of peptide bonds
b. organic solvents interfere with R-group interactions because these solvents also can form
hydrogen bonds; quickly denature proteins in bacteria, killing them
PRECIPITATION
By Acids- due to presence of the NH2 group in their molecules, amino acids have basic properties and form
insoluble salts with acids.
Organic acids- Trichloroacetic, Phosphomolybdic, Phosphotungstic, Picric, Tannic acids are used as
precipitants
• Tannic acid & Picric acid are used for treating burns because they produce astringent effect on
the tissues, diminish secretion of mucous membranes and prevent absorption of toxins. They are
also administered in some forms of gastro-intestinal irritation to relieve diarrhea.
• Inorganic acids also precipitate protein. Nitric acid is used for detecting the presence of proteins
in the urine (Heller’s test).
By salts of Heavy metals (alkali metals)- Hg, Ag, Pb

proteins behave like acids by virtue of the carboxyl radical; form insoluble salts with alkaline metals
• In metallic poisoning, antidotes like egg white and milk are given. They form insoluble precipitates
which can be removed subsequently by means of a stomach tube.
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By Alcohol- Maximum precipitation occurs at isoelectric point (when proteins are electrically neutral). This
accounts for the antiseptic property of alcohol.
* 95% alcohol has less germicidal property compared to 70% alcohol. It readily coagulates proteins and
coagulum forms a protective coating on the surface of bacteria preventing further penetration of the alcohol.

1. Which structural levels of a protein are affected by denaturation?
________________________________________________________________________________________
________________________________________________________________
2. In what way is the protein in cooked egg the same as that in a raw egg?
________________________________________________________________________________________
________________________________________________________________________________________
____________________________________________________.
3. Why is 70% ethanol rather than pure ethanol preferred for use as an antiseptic agent?
________________________________________________________________________________________
________________________________________________________________________________________
____________________________________________________.
Assessment
Given the table below, look for words that are related to protein. Classify if it’s a source, classification, property,
example or functions of proteins. (30 points)
P R O A M P H O T E R I C T E I N S A R E O R G A M I L K N
I C M A C R R O M O L E H C U B L G E M H A D E U P O F C A
R B O N H Y D O R O G E E N O E X L Y G E E N A N D N I T R
O G B E N W H I L C H F E U N E C O T I M E A T O N P R I W
M A R O I L Y F O A R B S O D F Y B B P O U L T R Y U I F O
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L D I N D G A N D T M I E S S U E U R E G P A I R A A S L R
W E C C L Y L A S A N I E N Z Y M L E H L O R M O C N E O K
A S N O D A B N T I B O N D I E S I I T O C A N B O E S C I
O O U N L R C U E D O U T E F R O N G M B A N I M N A L C N
A U N J D L A N I I M A L P S R L O L D I U C T S A N D U G
S R O U R C A E G L O B U L A R E S Y S N S K U C S H A L M
S C M G I L K G E G D G A L B U G M C I N I E C H E E S A O
E E C A K E S P E A S I T R I E U S O I L M R M E A L E T L
C O E T L I S H A N R A N I N E M B P A Y P A F Y F A G I E
O F N E S C H I L I E B A G S T E I R A C L T R N O W G O C
R E R D C H R I S T M A S Y A Z S Z O A H E I U K O I Y N U
I N A P R I S T I N E K E S M O D E T B E P N I O D N O L L
A E L R A B U B B L E G A I I L S I E M E R O T T S E L U E
N R B O L I P O P R O T E I N M A T I T H O E F I W T K R S
D G U T N C O T T O N K E L O R G A N I C T L L S Y I C E T
E Y M E A C A S E I N V I S A S O I S L U E B A S L G E J O
R A I I T R Y P S I N O G E C N H N A S A I L M U C L A N Z
M D N N Y A T H A B A S C A I C R I U S H N F B E A U R E T
Y M H S N E W Y E A R D A Y D O B E S I T S Y E R A T N N I
R T A L L D E N A T U R A T I O N O R T A N G B E E E P Z L
C S A N G Y O P S U M G R E E N C C Y S O A B E P C L I Y L
E O N E N U T R I T I O N O N E H F N H A N T E A I I N M A
N O N M A C R O N U T R I E N T I J E O O A E R I N N K E S
E G U M A M E L A J E L L Y N E C F A R I T H S R M S Y S J
B I O M O L E C U L E A M R I E O S E M R B A K I M B A B F
C H O L E S T E O L S Y M A N G O T O I I A W A S A B E L
N K I M C H I C R U S T O A C E A N S N A N T D C A N D Y A
R E T I N O L W A X E S S F I S H A N E D C H I P S R A Y N
R F P R E C I P I T A T I O N S H A R S M A I N N E U Z Z I
B I L E P E P S I N I M N O R L I F E H O U S E C H I T I N
SOURCES PROPERTIES EXAMPLES FUNCTIONS CLASSIFICATIONS
LABORATORY
INTRODUCTION
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Proteins are macromolecules with three dimensional structures called conformation. This 3D conformation of
the protein plays an important role in its function and solubility in a variety of solvents.
Several factors can affect this conformation; this includes temperature, extremes of pH, UV light, just to
mention a few. Proper adjustment to these factors can lead to the rearrangement of molecules in the protein
structure causing them to precipitate, coagulate or become denatured.
NAME: _____________________________ DATE: _____________________________

Activity No. 18
PROPERTIES OF PROTEINS
OBJECTIVES:

1. ascertain correctly the effects of acids, bases, inorganic salts, organic solvents and temperature on
the physical properties of proteins
2. appreciate completely the relationship of the structure of the protein to its function
MATERIALS
Reference material Pen/pencil

Scoring rubric
For Procedures:
o 1 point for the answer or written observation
For Rationalization
not clear
not clear
A. Solubility and Protein Denaturation
Effect of acids
1. Should you have 3 test tubes with albumin, casein and gelatin and you added 2 drops of 3M HCl, what
would be the possible result? What will happen when you add 5 more drops to the same test tubes?
How about when you add 10 more drops? Write the possible observation on the table below and
download a photo of the positive result. Attach these photos to your paper.
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Observations:
3M HCl
Initial appearance 2 drops 5 drops 10 drops
2% albumin
2% casein
2% gelatin
How will the 3M HCl affect each protein?

__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
Effect of bases
1. Should you have 3 test tubes with albumin, casein and gelatin, what would be the possible result when
you add it with 5 drops of 3M NaOH? How about when you add 10 more drops of NaOH? Write the
possible result on the table below. Download a photo of the positive result and attach it to your paper.
Observations
3M NaOH
5 drops 10 drops
2% albumin
2% casein
2% gelatin
Effect of heavy metals

1. Should you have 3 test tubes filled with albumin and each will be added with 0.1M CuSO 4 to the first
tube, 0.1M AgNO3 to the second tube and 70% isopropyl alcohol to the third tube, what would be the
possible result? Write your answer on the table below. Download a photo of the positive result and
Test tubes 0.1M CuSO4 0.1M AgNO3 70% isopropyl alcohol

1
2
3
How will the metal salts affect albumin?

__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
How will the isopropyl alcohol affect the albumin?

__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
B. Salting Out with Ammonium sulfate
1. Should you have 2% albumin in a beaker and you added 25 mL of saturated ammonium sulfate
solution, what would be the possible result?
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
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2. Supposed you filter the solution and collect the filtrate and the precipitate and you did the following:
➢ Place 2 mL of 2% albumin to the first test tube
➢ Place 2 mL of the filtrate into the second test tube
➢ Place a small portion of the precipitate into the third test tube and then add 2 mL of distilled
water. Swirl to mix the contents.
And then you add 10 drops of 3M NaOH into each of the test tubes followed by 5 drops of 0.1M
CuSO4, what would be the possible result for each test tubes? Write your answer below. Download a
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
C. Effect of Heat
How do heat affect albumin?
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________

Scoring Rubric
1. What are the differences between the different levels of protein structure
2. Differentiate coagulation, precipitation and denaturation
3. What is “salting in” and “salting out” in relation to proteins
4. What is the importance of being able to separate protein fractions in a sample?
5. How do heavy metal ions precipitate proteins from solutions?
6. A person accidentally swallowed a poison believed to contain heavy metals. Suggest a first aid method.
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Week no. 13
SUBMODULE NO. 13
PROTEIN
METABOLISM
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LESSON: Protein Metabolism
TOPICS:
1. Nitrogen Equilibrium
2. Biosynthesis of Proteins
3. Biosynthesis of NPN Compounds and Urea
4. Degradation of Amino Acids -Decarboxylation -Transamination -Oxidative deamination
5. Metabolism of the Carbon Skeletons of Amino acids
5.1. Ketogenic Amino Acids
5.2. Glucogenic Amino Acids
5.3. Clinical Significance of Protein Metabolism
TIME/SCHEDULE: WEEK 15
LECTURE: 5 hours
LABORATORY: 4 hours
LEARNING OBJECTIVES
1. Discuss the metabolism of amino nitrogen;
2. Describe the metabolism of the different amino acids;
3. Discern the importance of the knowledge of metabolism in keeping oneself healthy;
4. Appreciate the significance of the metabolism of different biomolecules in the maintenance of normal
life processes;
5. Internalize the relationship that exists among the different metabolic pathways;
6. Develop a persevering attitude in studying intricate metabolic processes;
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LESSON PROPER:
INTRODUCTION
An animal is said to be in a biological steady state when the composition and quantity of the diet are such
that no significant changes are occurring in the composition of the tissues of the body or in the total body weight.
All the proteins in the body are constantly breaking down to their constituent amino acids and being
resynthesized. The body proteins are said to be in a dynamic state. The rate of turnover varies from different
proteins in different tissues. Turnover rate of a compound is the percentage of the amount present which is
metabolized per unit or it may be expressed in terms of mass of compound transformed per unit weight of tissue
or animal. (Bernaldez & Hessari, 2001)
Lesson 1. Protein Metabolism
Activity 1. Given the following figure, answer the question that follows:
Stoker, H. Stephen. Biochemistry, 1st Philippine Reprint 2011.

Based on the information in the previous figure, determine the location within the body where each of the
following aspects of protein digestion occurs:
1. The enzyme aminopeptidase is active _______________________________

2. Amino acids are produced _______________________________
3. Peptide bonds are hydrolyzed under the action of trypsin _______________________
4. Large polypeptides are produced __________________________
5. Protein is denatured by HCl _____________________________
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PROTEIN METABOLISM
Protein forms the primary constituent not only of the cell but also of such regulatory agents as hormones
and enzymes. It confers upon the tissue a sort of biological specificity; that is, the protein of a particular tissue is
distinct and different from those of other tissues. This is so because complex proteins are made up of varying
proportions of essential and non-essential amino acids depending upon the tissue, organ or specie.
ESSENTIAL OR INDISPENSABLE AMINO ACIDS:

- Methionine - Leucine
- Threonine - Phenylalanine
- Lysine - Tryptophan
- Valine - Histidine
- Isoleucine - Arginine
The term essential and non-essential relates only to dietary requirements, not in relation to importance
in metabolism.
The study of protein metabolism is centered on the metabolism of amino acids, mostly concerned with
the fate of the amino nitrogen and those of the non-nitrogenous residues.
The amino nitrogen enters in the formation of urea while the non-nitrogenous residues participate either
in the carbohydrate or lipid metabolism.
ABSORPTION OF AMINO ACIDS

After a complete hydrolysis in the gastro-intestinal tract, the proteins are split into their component amino
acids. These amino acids are absorbed mainly in the small intestines through the portal circulation, and only
slightly through the lymphatics. The absorption of amino acids is very rapid so that the maximum concentration
in the blood is attained 30 to 50 minutes after eating.
There is rapid removal of the absorbed amino acids from the blood. The amino acids are quickly disposed
and appear in all tissues and organs of the body. This is evident from the fact that the amino acid nitrogen level
is kept more or less constant between 4-8 mg/100 ml of blood plasma.
FACTORS THAT AFFECT DISTRIBUTION OF AA IN METABOLIC REACTIONS

1. Convalescing or growing individuals – for constructions of new proteins
2. The availability of new carbohydrate and lipid - influence the amount of total caloric requirement which must
be supplied by amino acids
3. The pattern of amino acid mixture supplied to the tissues
4. The influence of hormones in modifying the direction and the rate of certain metabolic reactions
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Three sources of AA in the AA pool:
1. Dietary protein
2. Protein turnover – repetitive
degradation and resynthesis of protein
(degradation is from disease, injury and
wear and tear)
3. Biosynthesis of AA in the liver
Decarboxylation – elimination of the N
portion via urea; biosynthesis of NEAA
and NPNC compounds
Deamination – carbon portion: TAG via
FA biosynthesis; glucose via
gluconeogenesis; ATP via CAC, ketone
bodies via ketogenesis. Reamination
through reverse
Metabolic pathways of absorbed Amino acids

1. Synthesized into new tissue proteins including enzymes and hormones
2. Synthesized into plasma proteins
• Liver is the site for biosynthesis of plasma proteins, albumin, globulin and fibrinogen
Synthesis is dependent upon the presence of indispensable amino acids and is increased when there is
generous supply of dispensable AA.
3. Synthesized into liver proteins
4. Synthesized into non-protein nitrogenous components of various cells
• Purine, pyrimidine, porphyrins, glutathione, creatine, thyroxin, nicotinic acid, etc.
5. Decarboxylated with formation of physiologically active amines
6. Deaminated, with the subsequent formation of urea and oxidation of α-ketoacid or its conversion into
carbohydrate or fat
7. Take part in the glucose-alanine cycle
• Prevents the fluctuations in the blood glucose level between meals
• Functions
o To carry amino groups from skeletal muscles to the liver to be converted into urea for excretion
o To provide the working muscles with blood glucose made by the liver form the carbon structure
of alanine
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Lesson 2. Nitrogen Metabolism
DYNAMIC ASPECTS OF NITROGEN METABOLISM
NITROGEN BALANCE- quantitative difference between the nitrogen intake (from food) and the nitrogen output
(represented by nitrogen lost in the urine and feces)
POSITIVE NITROGEN BALANCE- intake exceeds the output; this is found whenever new tissues are being
synthesized as in growing young, convalescence, pregnancy, etc.
NEGATIVE NITROGEN BALANCE- output exceeds the intake; this is found in:
1. inadequate intake of proteins as in fasting, malnutrition, diarrhea;
2. increased catabolism as in fevers, infections, wasting diseases; and
3. increased loss of body proteins as in lactation, albuminuria
UTILIZATION OF INORGANIC NITROGEN

The only form of inorganic nitrogen, which can be utilized by living cells is ammonia.
Fixation of ammonia
1. Glutamic Acid synthesis
a-ketoglutaric acid + NH3 + NADH + H+ Glutamic acid + NAD+ + H2O
Enzyme: Glutamic acid dehydrogenase

• The α-amino group of glutamic acid can be transferred to other keto-acids by transamination process
• Provides a mechanism for synthesis of other amino acids
TRANSAMINATION
Is a biochemical reaction that involves the interchange of the amino group of an a-AA with the keto group
of an a-keto acid.
2. Synthesis of Glutamine
Glutamic acid + NH3 + ATP Mg++ Glutamine + ADP + Pi + H2O
Enzyme: glutamine synthetase
• Glutamine serves as a means of presenting to an enzyme an unprotonated nitrogen atom at reduction
level of NH3
• Glutamine serves as a store of ammonia
3. Carbamoyl Phosphate synthesis
CO2 + NH3 + ATP + H2O Carbamoyl PO4 + ADP + Pi + 3 H+
Enzyme: Carbamoyl phosphate synthetase
• Carbamoyl PO4 becomes available to the cells for carbamylation of amino groups as in the synthesis of
citrulline from ornithine
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FATE OF AMMONIA:
The ammonia liberated during the deamination process is disposed off in several ways:
1. It enters the general ammonia pool of the body and may be drawn upon either for anabolic or catabolic
purposes. It may be used in the reductive amination of keto acids derived from CHO to form new amino acids.
2. Since ammonia is toxic in large concentrations, it is being detoxified by synthesis to glutamine.
3. Ammonia may be excreted directly into the urine.
4. Ammonia may enter the ornithine cycle to form urea.
UREA CYCLE:
Mammals utilize urea as the major vehicle for excretion of surplus nitrogen (ureotelic). On the basis of
their observations, Krebs and Henseleit proposed a cyclic mechanism for the synthesis of urea involving the
amino acids ornithine, citrulline and arginine in the presence of the enzyme arginase, which catalyzes the
irreversible hydrolysis of arginine to ornithine and urea.
1. Nitrogen enters the urea cycle in the form of ammonium ion.

2. A reaction between ammonium ion and CO 2 produces Carbamoyl phosphate in a reaction that requires two
molecules of ATP for each molecule of carbamoyl phosphate.
3. Carbamoyl phosphate reacts with Ornithine to form Citrulline. (ornithine transcarbamoylase)
• The reactions of the cycle at this point take place in the mitochondrion.
• Citrulline is then transported to the cytosol.
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4. A second nitrogen enters the urea cycle when aspartate reacts with citrulline to form argininosuccinate
(arginosuccinate synthase) in another reaction that requires ATP (AMP and Ppi are produced in this reaction).
• The amino group of the aspartate is the source of the second nitrogen.
5. Argininosuccinate is split to produce arginine and fumarate. (argininosuccinate lyase)
6. Finally, arginine is hydrolyzed to give Urea (arginase) and to regenerate ornithine, which is transported back
to the mitochondrion.
7. The synthesis of fumarate is a link between the urea cycle and the citric acid cycle.
• Fumarate is an intermediate of the citric cycle, and it can be converted to oxaloacetate.
• A transamination reaction can convert oxaloacetate to aspartate, providing another link between the two
cycles
Degradation of Amino Acids for Energy Production

Five amino acids are degraded to Acetyl CoA by way of pyruvate
• Alanine
• Threonine
• Glycine
• Serine
• Cysteine
Five amino acids are degraded to acetyl CoA without forming pyruvate
• Lysine
• Tyrosine
• Phenylalanine
• Tryptophan
• Leucine
Five amino acids forming α-ketoglutarate

• Glutamate
• Glutamine
• Arginine
• Proline
• Histidine
Amino acids forming succinyl-CoA

• Methionine
• Valine
• Isoleucine
• Aspartate and Asparagine are deaminated to oxaloacetate
• Aspartate + α-ketoglutarate oxaloacetate + glutamate
• Asparagine + H2O Aspartate + NH4+
Activity 2. Supply the following figure with the proper amino acids required for each process.
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Assessment:
Test I: Identification. Identify what is being described. Write your answer legibly on the space provided before
each number. Answers with erasures will not be corrected.
______________________________1. What amino acid is considered to be dietary essential?

______________________________2. What structure of amino acid is responsible for its identity?
______________________________3. This is the structure of amino acid where the amino and carboxyl
groups are attached.
______________________________4. What amino acid is considered as basic apart from lysine and
arginine?
______________________________5. How many peptide bonds are present in a nonapeptide?
______________________________6. This is a type of amino acid where the amino group is attached to the
1st carbon adjacent to the carboxyl group.
______________________________7. This is the source of amino group in the synthesis of
argininosuccinate.
______________________________8. This is referred to as the repetitive process where protein is degraded
and resynthesized
______________________________ 9. TRUE or FALSE. 75% of the free AA in a healthy adult go into protein
synthesis to replace old tissue and build new tissue.
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______________________________10. When the protein of a particular tissue is distinct and different from
those of other tissues, what is this called?
Test II: Matching type. Match column A with Column B. Write your answer in capital letters on the space
provided before each number
A. Symbols. Match the amino acid with their respective symbols
Column A Column b
_____11. Phenylalanine A. W
_____12. Threonine B. T
_____13. Tryptophan C. F
_____14. Lysine D. G
_____15. Glycine E. K
F. T
G. P
B. Tests. Match the amino acid with the test specific for it.
_____16. Histidine A. Pauly’s reaction

_____17. Tryptophan B. Millon’s test
_____18. Cysteine C. Sakaguchi reaction
_____19. Tyrosine D. Nitroprusside test
_____20. Arginine E. Aldehyde reaction
F. Ninhydrin Test
G. All of these
H. None of these
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LABORATORY
INTRODUCTION
The term chromatography was originally applied by Michael Tswett, a Russian Botanist, to a procedure
where a mixture of different colored pigments (chlorophylls and xanthophylls) was separated from each other. It
was originally applied to separate colored substances but with a simple technique can also be used to separate
colorless substances such as amino acids.
Chromatography involves a sample (or sample extract) being dissolved in a mobile phase (which may be a
gas, a liquid or a supercritical fluid). The mobile phase is then forced through an immobile, immiscible stationary
phase. The phases are chosen such that components of the sample have differing solubilities in each phase. A
component which is quite soluble in the stationary phase will take longer to travel through it than a component
which is not very soluble in the stationary phase but very soluble in the mobile phase. As a result of these
differences in mobilities, sample components will become separated from each other as they travel through the
stationary phase.
The relative degrees of separation of the components of a mixture may be expressed by comparing their rate
factors or ratio of front or Rf values. To compute the Rf value, use the following formula:
Rf = Distance of compound
Distance of solvent
NAME: _____________________________ DATE: _______________________________

Activity No. 19
CHROMATOGRAPHY OF AMINO ACIDS

OBJECTIVES

1. identify correctly the importance of chromatography in biochemistry
3. State confidently the basic principles involved in this technique
MATERIALS

Scoring rubric
For Procedures:
For Picture
de
- 5 – outstanding – the photo/documentation shows genuine effort on the part of the maker and all parts are labelled with no
misspelled words
- 4 – excellent – the photo/documentation shows above average effort on the part of the maker, 1 or 2 parts are not labelled
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- 3 – Good – the photo/documentation shows average effort on the part of the maker, 3 to 4 parts are not
- 2 – Fair – the photo/documentation shows below average effort on the part of the maker, 5 or more parts
- 1 – Poor – the photo/documentation shows little effort on the part of the maker, and the illustration is not
labelled
The following are procedures in the performance of chromatography for amino acids. While it’s true that the
said procedures cannot be performed, it is still advised that you should be familiar with the steps. Read through
the procedures then proceed to the table that needs to be completed.
Preparation of the Chromatogram

1. Obtain a filter paper and cut it into 12 x 24 cm rectangle
2. Roll and see if it will fit a 400 mL beaker. Make the necessary adjustments if not.
3. Pull the filter paper from the beaker and place on top of a clean paper.
4. Using a pencil, draw a horizontal line 2.5cm from the bottom of the filter paper. This will serve as your
starting point
5. Draw 5 circles of approximately 2-3 mm in diameter on the horizontal line. The circles should be at least 2
cm apart.
6. Choose 5 amino acids from the counter and place 0.5 mL in a small test tube.
7. Using a capillary tube, apply the available amino acids on the corresponding spots. Let the spot dry first
before doing a second application. One amino acid is applied per spot. Use separate capillary tubes per
amino acid. Be sure to label everything properly using pencil.
Preparation of Developing Chamber

1. Place butanol-ethanoic acid mixture to a height of 1 cm in a 400 mL beaker
Chromatography proper
1. Get your chromatogram and roll it in to a cylinder. The edges should be touching end to end. Secure the
edges with a scotch tape
2. Carefully place the chromatogram in the beaker. The sample spots should not touch the solvent and be sure
not to touch the side of the beaker when putting your chromatogram inside.
3. Cover the beaker with parafilm and let it stand undisturbed for an hour or until the solvent front is 1 cm from
the top of the chromatogram.
Document your set-up and label with ink, to be passed together with the questions for research
Measurement and Computation of the Rf

1. When the solvent front is already 1 cm from the top of the chromatogram, remove it from the beaker.
2. Remove the scotch tape and spread it on top of a clean paper
3. Immediately mark the solvent front with a pencil at 5 points corresponding to the circles found at the bottom
of the chromatogram
4. You may air dry the filter paper or use an oven or hair drier. If heat will be used, you have to watch it since
butanol is flammable.
5. After drying, spray with ninhydrin and wait for spots to develop.
6. Mark the center of the spot with a pencil
7. Measure the distance travelled by the solvent and record on the data sheet below
8. Measure the distance travelled by the individual amino acids and record on the data sheet below.
9. Research on the literature rf of the different amino acids indicated: phenylalanine, tyrosine, tryptophan,
arginine, and methionine. Research as well on the expected color of spot formed. Download a photo of the
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Chromatography data sheet
Amino acid Literature Rf Color of spot
Scoring Rubric
1. State the principle behind chromatography.

2. What is the importance of chromatography in protein analysis?
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WEEK NO. 14
SUBMODULE NO. 14
LIPIDS:
CLASSIFICATION,
PROPERTIES AND
DIGESTION
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LESSON: LIPIDS
TOPICS:
1. Definition and Functions of Lipids
2. Classification of Lipids
2.1. Derived Lipids
2.2. Simple Lipids
2.3. Complex / Compound Lipids
2.4. Miscellaneous Lipids
3. Properties of Lipids
4. Digestion and Absorption of Dietary Lipids
LECTURE: 5 hours
LABORATORY: 4 hours

1. Define the different terms encountered in Biochemistry
2. Classify the different lipids correctly
3. Differentiate the properties of lipids from other biomolecules
4. Trace accurately how lipids are digested
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LESSON PROPER
INTRODUCTION:
LIPIDS
- Applies to a class of compounds that are soluble in organic solvents and nearly insoluble in water
- Chemically, lipids are either compounds that yield fatty acids on hydrolysis or complex alcolipids that
can combine with fatty acids to form esters
- Some lipids also contain non-lipid groups such as sialic, phosphoryl, amino or sulfate groups, which
tend to increase their solubility
- They are widely distributed in nature. True fats generally constitute the storage material for energy in
both plants and animals
o They are abundant in the subcutaneous and intramuscular connective tissues, the omentum,
etc., where they serve as heat insulator and reserve supply for energy
- Compound fats however are more often closely associated with the vital processes
o The cerebrosides for example are found only as constituents of highly specialized brain and
nervous tissues
o The phospholipids and sterols along with proteins and carbohydrates to the colloidal structure of
living matter
Functions of Lipids
1. Serves as hormones and hormone precursors

2. Aids in digestion
3. Provides energy storage and metabolic fuels
4. Acts as functional and structural components of biomembranes
5. Forms insulation to allow nerve conduction or to prevent heat loss
CLASSIFICATION OF LIPIDS
I. Simple lipids – ester of fatty acids with various alcohols

- Neutral fats – glycerol esters
- Waxes – esters of higher alcohols
o True waxes
o Cholesterol esters
o Vitamin A esters
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o Vitamin D esters
II. Compound lipids – those containing other radicals

- Phospholipids – contain H3PO4 and nitrogenous base
o Lecithin – choline as nitrogenous base
o Cephalin – ethanolamine
o Lipositol – inositol
o Phosphatidyl serine – serine
o Plasmalogen – resemble lecithin and cephalin in structure
o Sphingomyelin – sphingosine and choline
- Glycolipids – contain carbohydrates and nitrogenous base
- Ill-defined lipids – amino lipids and sulfolipids, having amino and sulfate groups respectively
III. Derived lipids – products of hydrolysis of I and II; but still exhibiting the general physical characteristics of
lipids
- Saturated and unsaturated fatty acids
- Mono and di-glyceride
- Alcohols
o Straight chain – products of hydrolysis of waxes
o Alcohols containing the b-ionone ring – vitamin A and some carotenoids
o Sterols
IV. Miscellaneous
- Aliphatic hydrocarbons – iso-octadecane from liver
- Squalene – hydrocarbons in shark liver and human sebum
- Carotenoids
- Vitamin D, E and K
FATS
- They are neutral ester of monobasic fatty acids with trihydric alcohol, glycerol. The general structure is:
- When R1, R2 and R3 represent the same fatty acid chain, the compound is a simple glyceride
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- More often however R1, R2 and R3 represent
different fatty acid chains and the compound
is called a mixed glyceride. Naturally existing
fats are usually of this type
- Naturally existing fats are usually of this type
by rearrangement of the fatty acid chains in
the fat structure a variety of glycerides result.
This helps to explain the presence of different
fats in the different organs of animals of the
same species
https://epomedicine.com/medical-students/structure-of-fatty-acids-and-derivatives-simplified/
Animal fat – contains mostly oleic, palmitic, and stearic acids

Mutton fat – has more stearic and less oleic than pork
Butter fat – composed mainly of palmitic and oleic acids with small amount
of butyric acid and caproic acid
Human fat – mostly oleic acid; has a yellowish tinge due to carotene and
xanthophylls pigments derived from the ingested food
Hydrolysis of Triglyceride
https://www.researchgate.net/figure/Fig-depicts-the-hydrolysis-of-triglyceride-to-glycerol-and-fatty-acids_fig2_317264701
- The hydrolysis of triglycerides is similar of ester hydrolysis wherein water (or a strong base) cleaves the
ester bond releasing the alcohol (glycerol) and carboxylic acids (fatty acids)
Fatty Acids
- These are products of fat hydrolysis
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1. Saturated Fatty Acids
- These belong to the acetic series and have the general formula of CnH2nO2
- Palmitic acid make up about 15 to 50% of the total fatty acids in fats. Myristic and stearic acids are also
present
- Tuberculo stearic (10-methyl stearic acid) acid occurs in the lipids of human tubercle bacilli
- They are abundant in butter, animal fat, lard, coconut and peanut oil
o Saturated fatty acids having low MW (10 or less C atoms) are liquid at ordinary RT. They have
low MP and are volatile. The rest are solids, with higher MP and are non-volatile. Those with 4
or less C atoms are miscible with water in all proportions
2. Unsaturated Fatty Acids

- The degree of unsaturation varies with the number of double bonds present in their structure
- They are unstable and reactive due to the presence of the double bonds in their molecules. The
reactivity increases with the increase of the double bond
- They are insoluble in ordinary solvents
- Liquid at RT and are non-volatile
- The greater the degree of unsaturation, the lower are the melting and congealing points (more fluid)
o Oleic acid is the most abundant fatty acid found in nature (about 50% of the total fatty acid in
many fats)
▪ Found in all naturally existing fats and phospholipids
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o In the single unsaturated fatty acids of animal lipids, the double bond in generally in the 9,10
position
OLEIC ACID
- The presence of the double bond gives rise to the possibility of geometric isomerism called cis-trans
isomerism
- If the radicals are on the same side of the bond, the compound is called a “cis” form, and at the
opposite is called “trans” form
- Common unsaturated fatty acids (with 2 or more double bonds)

o Linoleic acid*
o Linolenic acid*
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o Arachidic acid* *essential fatty acids
- Essential/indispensable fatty acids (EFA)

o Those that cannot be synthesized in the human body and must be supplied by the diet
o The most abundant EFA in mammals is the multiply unsaturated fatty acid linoleic acid
o The absence of linoleic acid and the rest of the EFA in the diet produces symptoms of EFA
deficiency such as sparse hair, scaly skin, and poor healing of wounds. Arachidonic acid is also
classified as an essential FA although the body can synthesize this substance from linoleic acid
o Deficiency in EFA in the diet of rats produce impairment of growth, some form of dermatitis,
excessive thirst and in advanced cases, even kidney damage
o Some cases of eczema and dermatitis caused by deficiency of vitamin A are cured by the
addition of these EFA in the diet
o The required amount of EFA in the diet is 2% of total caloric intake for adults and 3-4% for
infants
3. Hydroxy and Cyclic acids
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- Chaulmoogra oil has attracted the attention of medical practitioners because the ethyl esters and
soldium salts of hydnocapric and chaulmoogric acid are used in the treatment of leprosy
- Phthioic acid has been shown to produce proliferation of epitheloid and giant cells and is probably the
agent responsible for the manifestation of tuberculosis
4. Polyunsaturated Fatty Acids (PUFA)

- Have a carbon chain in which two or more carbon-carbon double bonds are present
- Omega Fatty Acids
o Fish and fish oil are the most concentrated source but maybe found in some plants
▪ eicosapentaenoic acid – EPA
▪ docosapentaenoic acid – DPA
▪ docosahexaenoic acid – DHA
- they lower plasma triglycerides
- they have beneficial effects on coronary heart disease because of their thrombotic action (they reduce
platelet aggregation and blood clots)
5. Eicosanoids
- The biochemical derived from the fatty acid arachidonic acid
- Prostaglandins are the best known of the eicosanoid class. Which also includes the leukotrienes,
prostacyclins and thromboxanes
- Cell membranes release arachidonic acid in response to a variety of circumstances, including infection
and allergic reactions
- Prostaglandins
o Lipids that contain 20 carbon atoms including a five membered ring structure
o First isolated from seminal fluid and from the prostate gland (originally thought to be the only
prostaglandin, a single substance secreted by the male genital tract)
o Many different prostaglandins were found to be present in almost all animal tissues
o According to one theory, aspirin kill pain by inhibiting the synthesis of prostaglandin (PG)
The labels in parenthesis indicate specific

types of PG. Linoleic acid leads to PG (E1), a
potent fever-inducing agent, via a similar
pathway
Arachidic acid prostaglandin synthetase PG(G2) and PG (H2)

(Pain causing)
PG(G2) + PG (H2) prostaglandin synthetase PG (E2) + PG (F2)
(Fever Producing)
o They regulate a number of biological activities and thus exhibit hormonal behavior (e.g. known to
cause smooth muscles to contract)
o A clinical use for some prostaglandins is to induce labor
o Prostaglandins and some of their precursors are also thought to cause inflammation. The anti-
inflammatory, antipyretic and analgesic actions of aspirin and other non-narcotic analgesics are
believed to result from their ability to interfere with the synthesis of prostaglandins.
o The non-narcotic analgesics inhibit the action of the enzyme prostaglandin synthetase thus
reducing the formation of excess prostaglandins and the pain, fever, and inflammation
- Thromboxane
o Formed by platelets in the blood stream, which act as vasoconstrictors and stimulate platelet
aggregation as an initial step in blood clotting
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- Leukotrienes
o Formed by a variety of white blood cells as well as other tissues and cause many symptoms
associated with an allergy attack
Group of drugs used for eicosanoid control
Activity: Describe how the following drugs control eicosanoid production and name specific examples
for each group of drugs.
1. NSAID (non-steroidal anti-inflammatory drugs) –

2. COX-2 inhibitors –
3. Aspirin –
4. Cortisone –
Glycerol
- Simplest trihydric alcohol commonly known as glycerine

- Oily, colorless, heavy liquid with sweet taste
- By-product in the manufacture of soap
- Component of fat responsible for the (+) acrolein test (easily detected by its characteristic acrid odor)
USES OF GLYCEROL
- Widely used both for pharmaceutical and cosmetic preparation due to its solubility, hygroscopic nature
and solvent action
- When glycerol undergoes hydrolysis under alkaline conditions, it is split into glyceride and
dihydroxyacetone
- It can reduce Cu++ to Cu+
- Positive for Benedict’s and Fehling’s test
- when treated with HNO3, it forms nitroglycerine which is used in making dynamites and smokeless
powders
o used in medicine as vasodilation drug in the treatment of hypertension
o in the body, glycerol is liberated during digestion of fat, when free glycerol is absorbed and
metabolized, it yields a caloric value similar to glucose
General Properties of Fats
Physical Properties
Neutral fats
- characteristic greasy feel and penetrates through paper producing a translucent spot
- odorless, tasteless and colorless (when pure)
- color due to pigments present
- insoluble in ordinary solvents
- soluble in organic solvents (chloroform, benzene, ether, hot alcohol)
- non-volatile, produce characteristic crystals with definite melting point
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- neutral fats containing large amounts of unsaturated fatty acids are liquid at room temperature, hence
they are called oils. In the presence of catalysts (finely divided nickel) take up hydrogen and become
solid
CH3(CH2)4CH=CHCH2CH=CH(CH2)7COOH + H2 Ni CH3(CH2)16COOH
- principle involved in the manufacture of artificial butter (oleomargarine) from vegetable oils
Fats and Oils
- In general, fats are solid are room temperature and have a relatively high percentage of saturated fatty
acids and have a relatively high percentage of unsaturated fatty acid residues
- Oils which are liquids at room temperature have relatively high percentage of unsaturated fatty acid
residues
- Fats are obtained from animal sources and oils are obtained from plant sources
- Fats may be considered to be triesters formed from the glycerol and three molecules of fatty acids
- Most of the fatty acids in these esters have 14-18 carbons
- Because there are three ester groups per glycerol, the molecules are called triacylglycerols, or
triglycerides
Fat floats on water because it has a lower specific gravity than the
latter. When shaken vigorously with it, fats broke up into fine
particle forming a temporary emulsion. Soap, acacia, albumin or
bile salts are used as emulsifying agents (lower the surface
tension)
Chemical Properties
1. Hydrolysis
- the hydrolysis of a fat molecule generates 3 molecules of fatty acid a glycerol molecule
- catalysts: acids, enzymes (lipases), superheated steam
Stearin + 3H2O + acid/enzyme 3 stearic acid + glycerol

2. Saponification
- Base-catalyzed hydrolysis of a fat, when the base is NaOH, the sodium salts of the fatty acids are
produced
https://byjus.com/chemistry/saponification/
- Sodium soaps – hard
- Potassium soap – soft
- Ca and Mg – insoluble soap
- Detergency – property of the soap of causing and maintaining the emulsification of the greasy
substances
- The cleansing property of soap is attributed to its emulsifying property
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3. Hydrogenation
- Unsaturated fats (oils) are generally liquids. It is possible to obtain solid, saturated fats by catalytic
hydrogenation of oils
http://www.docbrown.info/page04/OilProducts14.htm
4. Rancidification
- Fats are neutral in reaction, but when exposed to air for some time, they become acidic due probably to
hydrolysis which results in the liberation of volatile fatty acids. The subsequent oxidation of the fatty
acid chains with the formation of odoriferous volatile aldehydes and ketones
Factors that contribute to the rapid onset of rancidity

o Enzyme
o Heat
o Light
o Moisture
o Bacteria
- Fats containing oleic acid become rancid quite easily
- To prevent rancidity in products containing unsaturated fats, compounds are added to serve as
antioxidants (e.g. vit. E and phenols)
- Rancidity results in the destruction of the accessory food factors like carotene, Vit. A and vit. E. Rancid
fat therefore s not only unpalatable but may even be toxic
5. Halogenation
- The halogens chlorine or bromine readily add across the double bonds of unsaturated fatty acids
- Halogenation of an unsaturated fat is used to estimate its degree of unsaturation. The IODINE
NUMBER is expressed in terms of the number of grams of iodine that would be absorbed by 100g of fat
Waxes and Terpenes
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Lipid waxes
- Esters formed from long-chain fatty
acids and high molecular weight
alcohols
- Examples
o a. Spermacetic wax
▪ Found in the head of
sperm whale, soft wax used in ointments
o b. Carnauba wax
▪ Found on Brazilian palm leaves, used in polishes and mimeograph stencils
- In nature, waxes serves as protective coatings on leaves, skin, feathers, fur and the outer skeletons of
some insects
Lanolin
- A mixture of waxes derived from wool; used as a base for ointments, salves and creams
Terpenes
- Compounds containing isoprene fragments
- Classified as lipids because they are biological molecules that dissolve in non-polar solvents
- No fatty acid residues in their structures
- Vitamin A and its precursors – B- carotene are terpenes
STEROIDS
- Derivatives of the hydrocarbon ring system
- One type of steroid found in living cells is a STEROL, a
compound in which a hydroxyl (OH) group is attached to
the steroid ring system
Cholesterol
- Most abundant sterol in animal tissues
- Present in nerve tissue, blood and bile (the greenish fluid
secreted by the liver that aids in the digestion of fat)
- Greek “chole” for bile and “steros” for solid
- Gallstones are nearly pure cholesterol (80 – 90%)
- In the body, cholesterol is the precursor of biological
steroids/steroid hormones which perform crucial body
functions (e.g. bile salts, androgen, estrogen, etc)
7-dehydrocholesterol
- Cholesterol derivative which is converted to an active
form of vitamin D (Vitamin D3 or cholecalciferol) when
skin is exposed to sunlight
https://www.sigmaaldrich.com/catalog/product/sigma/30800?lang=en&region=US
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FAT-SOLUBLE VITAMINS
Vitamins
- Organic compounds needed for normal growth and maintenance
2 major Classes
- Water soluble vitamins
o Vitamins that are soluble in water, can be excreted in the urine
o Not stored in the body inappreciable amounts and must be constantly applied in the diet
o Vitamin C and the B complexes
- Fat-soluble vitamins
o Absorbed along with dietary fats and are not excreted in the urine
o The body can store fat-soluble vitamins in moderate amounts and is thus not so dependent
upon daily supply of them in the diet
o Vitamin A, D, E, K
DERIVED LIPIDS
https://ibiologia.com/phospholipid-bilayer/
1. PHOSPHOLIPIDS
- Type of lipid found in all biological
membranes
- Composed of
o 2 fatty acids
o 1 glycerol
o 1 phosphoric acid
o 1 alcohol
Important Phosphoglycerides
Cephalin
- ethanolamine phosphoglyceride; phosphatidyl
ethanolamine
- The A (Alcohol) group in the formula is
derived from ethanolamine, HOCH2CH2NH2 (or cholamine)
- Found in brain tissue
- Cephalin is the thromboplastic substance which initiates the process of blood coagulation
http://www.tutorsglobe.com/homework-help/biochemistry/cephalin-71946.aspx
CEPHALIN
Lecithin
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- choline phosphoglyceride, phosphatidyl choline
- has a structure in which the A residue is derived from
the cation choline
- present in great quantities in the egg yolk, liver and
nervous tissue (arachidonic acid is one of its
component fatty acid)
- freshly prepared lecithin is waxlike, turns brown on
exposure to air; soluble in organic solvents and
yields acrolein when heated with dehydrating
agent
- 2 fatty acids + 1 phosphoric acid + 1 glycerol + 1
choline
- In the food industry, lecithins are used as emulsifiers
(agents that break-up mixtures of oil and water into an emulsion of tiny drops of oils suspended in water
- When only the unsaturated acid chain (R’) is removed by

hydrolysis from lecithin, a lecithin derivative known as lysolecithin forms
- Lysolecithin is a toxic substance that causes destruction of RBC
(hemolysis). Some snake venoms and insect poisons are toxic because
they contain enzyzmes capable of catalyzing the hydrolysis of lecithin to
lysolecithin
http://osp.mans.edu.eg/medbiochem_mi/cources/biochemistry/1st_year_dentistry/Basic_chemistry/lipid_chemistry/Files/Lecture_02.htm
Sphingolipids
- Also found in all membranes are particularly abundant in brain and nerve tissue
- Sphingomyelin and glycolipids are both classified as sphingolipids because they include a backbone of
sphingosine, an amino alcohol
Sphingosine
www.lipidhome.co.uk/lipids/sphingo/lcb/index.htm
- Replacing one of the amino H with a fatty acid acyl group R-
CO produces an amine of sphingosine known as ceramide
(basic structural units for all sphingolipids)
Ceramide
https://en.wikipedia.org/wiki/Ceramide
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-The myelin sheath that wraps nerve fibers contains phospholipids and
sphingolipids. In multiple sclerosis, lipids are lost from the sheath, which
leads to nerve damage
https://beyondthedish.wordpress.com/tag/myelin-sheath/
Sphingomyelin
- The most abundant sphingolipid found in the tissue of higher animals
- Contain PO4 groups
thus they have polar as
well as nonpolar
segments and
properties that are
similar to
phosphoglycerides
2. GLYCOLIPIDS
- Also known as “Sugar lipids” https://in.pinterest.com/pin/212654413633150198/
- Also include the ceramide (sphingosine + fatty acid)
structure, but their polar heads are composed of CHO
group in place of the phosphate ester of sphingomyelins
1. Cerebroside
- A glycolipid found in the membranes of brain tissue as its
name suggests
2. Ganglioside
- Found on the outer surface of nerve cells
- Glycolipids in which the CHO that is attached to ceramide
is musch more complex than a monosaccharide
- Tay-Sach’s disease
o the enzyme (hexosaminidase A) needed to break down a ganglioside is deficient leading to the
accumulation of the latter in the brain and spleen.
o This accumulation leads to neurological deterioration, which occurs after the first month of life
and leads to death within 5 years.
o a hereditary disorder
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o one of many genetic disorders in lipid metabolism that have been discovered in humans
DIGESTION AND ABSORPTION OF LIPIDS
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https://www.slideshare.net/AnupShamsherBudhatho/lipid-digestion-and-absorption
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Hydrolysis of fats may involve:
1. Complete hydrolysis – wherein triglycerides are hydrolyzed to glycerol and 3 fatty acids
https://www.researchgate.net/figure/Hydrolysis-of-triglycerides_fig6_253638502
2. Incomplete hydrolysis – happens when the fatty acid from the alpha – carbon of the glycerol is
removed resulting to the formation of B-monoglyceride and water.
https://guides.hostos.cuny.edu/che120/chapter11
Intestinal Absorption
- Complete hydrolysis is NOT a prerequisite to absorption. Fatty acid esters which are resistant to
hydrolysis are absorbed and deposited in the fat depot. Those which are not absorbed are also
completely hydrolyzed in the intestinal lumen and are excreted in the feces.
- Around 40% of ingested triglycerides are hydrolyzed to fatty acids and glycerol, 3-10% are absorbed as
triglycerides, while the rest is partially hydrolyzed to B-monoglycerides
- The glycerol portion, which is water soluble is absorbed through the portal route
- The fatty acid, which is water-insoluble is predominantly absorbed through the intestinal lymph (appear
in the form of triglycerides)
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- Answer activities listed in this unit
Scoring Rubrik for the short response assignment and case analysis
4 3 2 1
unsuccessful
accuracy
correctly cited
slows readability
ASSIGNMENT:
1. Olestra is an artificial fat substitute designed to allow individuals to obtain the taste and food
consistency of fat, without the calories from fat.
a. Draw and describe the structure of olestra
b. Describe how olestra is metabolized.
2. The mammary gland produces milk, which is the major source of nutrients for the breastfed human
infant. The fatty acid composition of human milk varies depending on the diet of the mother.
a. Differentiate the fatty acid content of human milk and cow’s milk (used in milk formula)
Although the concentration of pancreatic lipase and bile salts are low in the intestinal lumen of the
newborn infant, the fat of human milk is still readily absorbed.
b. Explain why this is so.
3. Will inhibiting pancreatic lipase help a person lose weight? Explain you answer.
Case:
Lucky Chan has several episodes of mild back and lower extremity pain over the last year, probably
caused by minor sickle cell crises. He then developed severe right upper abdominal pain radiating to his lower
right chest and his right flank 36 hours before admission to the emergency room. He states that the pain is not
Page 305 of 334

like his usual crisis pain. Intractable vomiting began 12 hours after the onset of these new symptoms. He
reposts that his urine is the color of iced tea and his stool now has a light clay color.
On physical examination, his body temperature is slightly elevated and his heart rate is rapid. The
whites of his eyes are obviously jaundiced. He is tender to pressure over his right upper abdomen.
The ER physician suspects that Lucky is not in sickle cell crisis but instead has with acute cholecystitis
or a gallbladder stone lodged in his common bile duct, causing cholestasis. His hemoglobin level was low at
7.6 mg/dL (reference range = 12 – 16) but unchanged from his baseline 3 months ago. His serum total bilirubin
level was 3.2 mg/dL (reference range = 0.2 – 1.0) and his direct bilirubin level was 0.9 mg/dL (reference range
= 0 – 0.2).
Questions:
1. If a gallbladder stone was lodged in the common bile duct of Lucky, what substances that aid in fat
digestion and absorption will be affected? Elaborate your answers.
2. If the stone stays in the common bile duct for prolonged periods of time. What possible deficiency will
occur. Explain your answer.
3. What is the possible cause of Lucky’s jaundice?
Page 306 of 334

LABORATORY
INTRODUCTION
The term “lipids” applies to a class of compounds that are soluble in organic solvents and insoluble in
water. Lipids contain carbon, hydrogen and oxygen but have far less oxygen proportionally than
carbohydrates.
Lipids are an important part of living cells. Together with carbohydrates and proteins, lipids are the main
constituents of plant and animal cells.
The triesters of fatty acids with glycerol makes up the class of lipids known as fats and oils. The
triglycerides are found in both plants and animals and compose one of the major food groups in our diet.
The steroids are one of the best known and best studied of the lipid groups, and example of which is
cholesterol. Cholesterol is amphiphilic having a polar head, represented by the –OH group and an extensive
nonpolar region made up of fused rings and hydrocarbon tails. The functional group of the cholesterol structure
is the hydroxyl group. Fatty acids are esterified at this position especially under physiological conditions.
Cholesterol and its ester derivatives are abundant in plasma proteins called lipoproteins whose function it is to
transport the cholesterol to peripheral tissue.
NAME: _____________________________ DATE: _______________________________

Activity No.20
PHYSICAL AND CHEMICAL PROPERTIES OF LIPIDS
OBJECTIVES

1. determine exactly the solubility of lipids in polar and non-polar solvents
2. perform precisely some tests that determine the presence of lipids in a sample
3. appreciate completely the importance of lipids in the body
MATERIALS
Test tubes Stearic acid Corn oil

Test tube rack Conc. Sulfuric acid Cholesterol powder
Bunsen burner Chloroform Olive oil
Test tube holder acetic anhydride Linseed oil
Test tube brush Chocolate Butter
Pasteur pipet Milk Lard
Watch glass Acetone Glycerol
Porcelain tile Raw egg yolk Potassium bisulfite
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Scoring rubric
For Procedures:
For Rationalization
o 3 – all concepts were mentioned and a clear hypothesis, generalization or conclusion was derived from the activity
o 2 – one or 2 concepts were not mentioned and the hypothesis, generalization or conclusion is not clear
o 1 – 3 or more concepts were not mentioned and the hypothesis, generalization or conclusion is not clear
Solubility and Physical Appearance: Watch: https://www.youtube.com/watch?v=l2QOi9mZoFc

1. Prepare 6 test tubes and label them properly: Corn oil, linseed oil, lard, butter cholesterol and stearic
acid
2. Place 5 drops of corn oil, and linseed oil to the corresponding test tube while a matchstick-head sized
portion of lard, butter, cholesterol and stearic acid will be used.
3. Note their physical state and write your observations on the data sheet below
4. Add 1 mL of water into each of the test tube
5. Shake and note their solubility
6. Write your observations on the data sheet below
7. Repeat procedure “a” and “b”
8. Add 1 mL of petroleum ether into each of the test tube
9. Shake and note their solubility
10. Write your observations on the data sheet below
11. Save the test tubes with ether for the next procedure
• You may want to try this at home by dissolving your mineral oil (baby oil), cooking
oil and lard in water.
pH
1. Prepare 5 sets of blue and red litmus paper and place them side by side on a watch glass or a
porcelain tile
2. Using the test tubes with ether from the procedure on solubility, test the pH of each of the lipids in the
test tube by adding a drop of each into the corresponding litmus paper
3. Record your results
Lipid Physical state Solubility in water Solubility in pH (using litmus

ether paper)
Corn oil
Linseed oil
Lard
Butter
Cholesterol
Page 308 of 334
Stearic acid
Make a general statement as to the solubility of lipids in various solvents:

________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
General tests
Spot test: Watch: https://www.youtube.com/watch?v=wpCtf2aqjs8

1. Prepare 3 test tubes and label them with the substances to be tested: chocolate, milk and cooking oil.
2. Place 5 drops of milk and corn oil to the corresponding test tube. For the chocolate, get a matchstick-
head sized portion.
3. Add 1 mL of acetone to the three test tubes and shake to dissolve the contents
4. Get one drop from the first tube containing corn oil and drop it on a 1/8 sheet of paper that is properly
labelled using pencil. Do the same for the other two test tubes using separate 1/8 sheets of paper.
5. Let the acetone dry
6. Hold the paper against the light and observe.
• Your may do this at home by placing a drop or a smear of oil onto a small piece of clean
paper and letting this stand for 5 minutes. Observe this against the light and take a photo
then attach this below this.
SPOT TEST
Observations:
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
Acrolein test: Watch: https://www.youtube.com/watch?v=l2QOi9mZoFc

1. Prepare 4 test tubes and label them with the substances to be tested: Glycerol, corn oil, lard and stearic
acid
2. Place 5 drops of glycerol and corn oil to the appropriate test tube while a matchstick-head sized portion
is used for the lard and stearic acid
3. Add a pinch of potassium bisulfate into each of the tube
Page 309 of 334

4. Carefully heat each of the tube until the potassium bisulfate is melted and slightly turns black. Do not
overheat since it would be difficult to clean the test tube afterwards!
5. Waft the odor from the test tube towards your nose. Do not inhale directly and note the odor produced
Observations:
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
______________________Which of the substances tested did not give a positive result?
___________________________________________________________________Why?__________
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
Test for degree of unsaturation: Watch: https://www.youtube.com/watch?v=_5ObG6fIAdQ

1. Get 4 test tubes and label them with the substances to be tested: Olive oil, corn oil, lard
2. Place 3 mL of chloroform into each of the test tubes
3. Add 5 drops of Lugol’s iodine
4. The solution should turn pink in color due to the presence of Iodine.
5. Get the test tube labelled as olive oil and add olive oil drop by drop until the pink color is discharged.
Count the number of drops. Repeat this procedure for the other test tubes.(melt a small amount of lard
in an evaporating dish, do not boil)
6. Observations:
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
______________________What is the principle behind this test?
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
Which of the substances is the most unsaturated?
__________________________________________________________________
Which of the substances is the least unsaturated?
__________________________________________________________________
Cholesterol
Caution: sulfuric acid is corrosive, be sure to wear goggles and gloves for this activity.
Leibermann-Burchard Test
1. Prepare 2 test tubes and label with the substances to be tested for: cholesterol and egg yolk
2. Place a pinch of cholesterol and a small amount of raw egg yolk in the appropriate test tube
3. Dissolve this in 0.5 mL of chloroform
4. Add 5 – 6 drops of acetic anhydride to each tube with gentle mixing
5. Carefully add 0.5 mL of concentrated sulfuric acid down the side of the test tube without mixing
Page 310 of 334

6. Immediately record and document your observations. Documentations should be passed together with
the questions for research.
__________________________________________________________________________________
__________________________________________________________________________________
Download a photo of the positive result for this test:
Salkowski test
1. Prepare 2 test tubes and label with the substances to be tested for: cholesterol and egg yolk
2. Place a pinch of cholesterol and a small amount of raw egg yolk in the appropriate test tube
3. Dissolve this in 0.5 mL of chloroform
4. Add 0.5 mL of concentrated sulfuric acid
5. Record and document your observations. Documentations should be passed together with the
questions for research.
__________________________________________________________________________________
__________________________________________________________________________________
Download a photo for the positive result of this test
Scoring Rubric
Page 311 of 334

1. Differentiate fats from oils.

2. What are glycolipids, phospholipids and sphingolipids? State their importance in the body
3. What is the importance of cholesterol in the body?
4. What is rancidity? What are the factors that bring about rancidity?
5. What is the Sudan III test?
6. What test can be used to differentiate saturated from unsaturated fatty acids. How is it done and what is
the positive result?
7. If the body cannot use carbohydrates as the source of energy, it will start using the fat reserves. What
are the advantages and disadvantages of this?
Week no. 17
Submodule no. 15
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Lipid metabolism
LESSON: LIPID METABOLISM
TOPICS:
1. Lipolysis
2. Knoop’s Beta – Oxidation of Fatty acids
3. Ketogenesis
4. Lipogenesis
5. Cholesterol metabolism
6. Clinical Significance of Lipid Metabolism
6.1. Obesity
6.2. Lipid Storage Diseases
TIME/SCHEDULE: WEEK NO 17
LECTURE: 5 hours
LABORATORY: 4 hours
LEARNING OBJECTIVES

1. Describe correctly terms associated with lipids;
2. Discuss correctly the metabolism of lipids in the body;
3. Correctly illustrate the pathways involved in the metabolism of fats
4. Differentiate accurately lipolysis from lipogenesis
5. Explain completely the importance of ketogenesis
6. Internalize thoroughly the relationship that exists among the different metabolic pathways;
Page 313 of 334

LESSON PROPER
INTRODUCTION
Fatty acids are a major fuel for humans and supply our energy needs between meals and during
periods of increased demand, such as exercise. During overnight fasting, fatty acids become the major fuel for
cardiac muscle, skeletal muscle and liver. The liver converts fatty acids into ketone bodies (acetoacetate and β
hydroxybutyrate), which also serve as major fuels for tissues (e.g. the gut). The brain, which does not have a
significant capacity for fatty acid oxidation, can use ketone bodies as a fuel during prolonged fasting
The route of metabolism of fatty acids depends somewhat on its chain length. Fatty acids are generally
classified as very long chain length fatty acids (greater than C20), long chain fatty acids (C12-C20), medium
chain fatty acids (C6 – C12), and short chain fatty acids (C4).
ATP is generated from oxidation of fatty acids in the pathway of β-oxidation. Between meals and during
overnight fasting. Long-chain fatty acids are released from adipose tissue triacylglycerols. They circulate
through blood bound to albumin.
LIPOLYSIS
In cells, fatty acids are converted to fatty acyl CoA derivatives of acyl CoA synthetases. The activated
acyl group is transported into the mitochondrial matrix bound to carnitine, where fatty acyl CoA is regenerated.
In the pathway of β-oxidation, the fatty acyl group is sequentially oxidized to yield FAD(2H), NADH, and acetyl
CoA. Subsequent oxidation of NADH and FAD(2H) in the electron transport chain, and oxidation of acetyl CoA
to CO2 in the TCA cycle, generates ATP from oxidative phosphorylation.
Steps of Beta-oxidation
Activation:
Page 314 of 334
- Once the triglycerides are broken down into glycerol and fatty acids they must be activated before they
can enter into the mitochondria and proceed on with beta-oxidation. This is done by Acyl-CoA
synthetase to yield fatty acyl-CoA.
- After the fatty acid has been acylated it is now ready to enter into the mitochondria.
- There are two carrier proteins (Carnitine acyltransferase I and II), one located on the outer membrane
and one on the inner membrane of the mitochondria. Both are required for entry of the Acyl-CoA into
the mitochondria.
- Once inside the mitochondria the fatty acyl-CoA can enter into beta-oxidation.
- Thiokinase – enzymes catalyzing the formation of fatty acyl CoA esters
- Found in the outer mitochondrial membrane
RCOOH + ATP + CoA RCO.CoA + AMP + Ppi

Transfer to Carnitine
- Higher fatty acids have limited ability to cross the inner membrane as CoA esters but their entry is
stimulated by carnitine
- Enzyme: Fatty Acyl CoA: Carnitine fatty acid transferase
RCO.CoA + Carnitine RCO.Carnitine + CoA
- The fatty acyl carnitine formed readily passes thru the inner membrane
Transfer to Intramitochondrial CoA
- The fatty acid is transferred from carnitine to intramitochondrial CoA

- Enzyme: Carnitine fatty acid transferase
Fatty acyl carnitine + CoA Fatty acyl CoA + Carnitine
- The mitochondria contain another type of acyl thiokinase participating in fatty acid activation. The
function of this enzyme (which is located in the matrix) is to make possible the direct utilization of GTP
generated in the mitochondria
Fatty Acid Degeneration
- After the activation, the fatty acyl CoA ester undergoes enzymatic degradation. The shortening of the
fatty acid chain by 2 C atoms is due to four successive steps
Oxidation: (Dehydrogenation)
- A fatty acyl-CoA is oxidized by Acyl-CoA dehydrogenase to yield a trans alkene. This is done with the
aid of an [FAD] prosthetic group.
- Formation of α,β-unsaturated derivative
Page 315 of 334

Hydration:
- The trans alkene is then hydrated with the help of Enoyl-CoA hydratase
- Reversible hydration – formation of 3-hydroxyacyl CoA
Oxidation: (dehydrogenation)
- the alcohol of the hydroxyacyl-CoA is then oxidized by NAD+ to a carbonyl with the help of
Hydroxyacyl-CoA dehydrogenase. NAD+ is used to oxidize the alcohol rather then [FAD] because
NAD+ is capable of the alcohol while [FAD] is not.
- Dehydrogenation involving NAD to yield the β-keto derivative
Cleavage:
- Finally acetyl-CoA is cleaved off with the help of Thiolase to yield an Acyl-CoA that is two carbons
shorter than before. The cleaved acetyl-CoA can then enter into the TCA and ETC because it is already
within the mitochondria.
- Reaction of β-ketoacyl CoA with CoA
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- yields acetyl CoA and a fatty acid derivative of CoA which is shorter by 2 Carbon atoms (Thiolysis)
https://biologydictionary.net/beta-oxidation/
Page 317 of 334

BETA-OXIDATION PATHWAY
Page 318 of 334

Ketogenesis and Ketosis
Ketogenesis https://www.nutritionadvance.com/ketogenesis-benefits-ketone-bodies/
- Formation of ketone bodies
- Takes place in the liver
- Under physiologic conditions, fatty acid degradation and
synthesis occur without significant accumulation of the
intermediates
Ketone bodies
- Acetoacetic acid
- Β-hydroxybutyric acid
- Acetone
Reactions:
https://www.sciencedirect.com/topics/medicine-and-dentistry/ketogenesis
Ketogenic
- Substance which form ketone bodies
- Example: fatty acids and the amino acid leucine, Isoleucine, phenylalanine and tyrosine
Antiketogenic
- Substances which prevent the formation of ketone bodies
Page 319 of 334

- Example: CHO, glycerol, fat, glucogenic amino acids (glutamic acid, glycine, alanine, aspartic acid,
serine, valine, cysteine, methionine, proline, ornithine, arginine, threonine, and histidine)
Ketonemia
- Accumulation of ketone bodies in blood
Ketonuria
- Presence of ketone bodies in urine when the blood level of ketone bodies exceed renal threshold
Fat Storage and Utilization

- Fats (triacylglycerols) are stored primarily in adipose tissue, which is widely distributed in the body.
- Fat is the major reserve of potential energy.
Lipogenesis
- The biosynthesis of fatty acids from acetyl-CoA is called lipogenesis.
- Acetyl-CoA can be obtained from the catabolism of carbohydrates, fats, or proteins.
- After they are synthesized, fatty acids combine with glycerol to form triacylglycerols, which are stored in
adipose tissue.
- Consequently, lipogenesis is the pathway by which all three of the major classes of nutrients are
ultimately converted to fat.
Lipogenesis vs. Beta oxidation

- Fatty acid catabolism occurs in the mitochondria, but fatty acid anabolism (lipogenesis) occurs in the
cytoplasm.
- Lipogenesis requires a set of enzymes that are different from the enzymes used in the catabolism of
fats.
- In the anabolic pathway (lipogenesis), the growing fatty acid chain bonds to a special acyl carrier
protein, ACP-SH, which acts as a ‘handle’ to transfer the growing chain from one enzyme to another
through the series of enzyme-catalyzed reactions in the pathway. Coenzyme A is the carrier in fatty
acid catabolism.
- A preliminary set of reactions, involving malonyl-CoA, occurs for each two-carbon addition cycle in the
synthesis. “Malonyl-” refers to a three-carbon group and has no counterpart in the catabolic pathway.
Page 320 of 334

COMPARISON OF LIPOGENESIS AND BETA OXIDATION
LIPOGENESIS
Page 321 of 334

Metabolism of Phosphatides
Phosphatide
- phosphoric acid
- Glycerol
- Fatty acids
- Nitrogenous base
- Absorption of Phosphatides
o Dietary phospholipids as well as those contained in digestive secretions may be absorbed as
such or by hydrolyzed by lecithinase from pancreatic juice
Page 322 of 334
Lecithin fatty acids + glyceryl-phosphoryl-choline
Cephalin fatty acids + glyceryl-phosphoryl-cholamine
- The glyceroPO4 linkage may be hydrolyzed I the intestinal mucosa by phosphodiesterase
Phosphoglyceride Synthesis
1. De novo synthesis - in which phosphatidyl serine serves as a precursor for other phosphatides
1,2 diglyceride + ATP Phosphatidic acid + ADP
Phosphatidic acid + CTP CDP diacylglycerol + Ppi
• CMP moiety is displaced by serine, inositol or glycerol PO4
CDP diglyceride + serine phosphatidyl serine + CMP
CO2
Phosphatidyl serine Phosphatidyl ethanolamine + CO2 Phosphatidyl choline
- Phosphatidic acid is an important intermediate in the synthesis of phosphoglycerides and TAG

- The phosphatidic acid itself may be formed from glycerol – 3 phosphate or DHAP
- The synthesis of glycerophospholipids can occureither by activation into CDP – choline and CDP –
ethanolamine or by formation of active diacylglycerol, CDP-diacylglycerol
Page 323 of 334

2. Utilization of exogenous choline (salvage pathway)
Choline + ATP Phosphoryl choline + ADP
CTP + phosphoryl choline CDP choline + Ppi
CDP choline + 1,2 diglyceride Lecithin
https://quizlet.com/30201442/e-tag-and-phospholipids-flash-cards/
Cholesterol Metabolism
- Similar to fats, the route of absorption is through the lymphatics, while cholesterol is readily absorbed,
its reduction products (formed through bacterial activity) coprosterol and cholestanol are poorly
absorbed and are found abundantly in the feces
- Cholesterol enters the intestines not only through the foods but also through the intestinal secretions
and bile
Biosynthesis of cholesterol
- Conversion of acetyl CoA to Mevalonic acid

- Conversion of mevalonic acid to squalene
- Conversion of squalene to cholesterol
Synthesis of cholesterol esters
Most of the cholesterol in the tissues is present in esterified form
Page 324 of 334

1. Cholesterol + Fatty acid CoA cholesterol ester + CoA
2. Phosphatidyl choline + cholesterol Lysophosphatidyl choline + Cholesterol ester
_________________________1. Reaction involved when acetic acid is converted to acetone
_________________________2. What is the 3-carbon atom group in lipogenesis that has no counterpart in
beta-oxidation.
_________________________3. Pancreatic enzyme that hydrolyzes dietary phospholipids
_________________________4. In the de Novo synthesis, what serves as the precursor to other
phosphatides?
_________________________5. In the salvage pathway, what nitrogenous base is used to produce
phosphatides?
_________________________6. What enzyme hydrolyzes the glycerol-PO4 linkage in phosphatides?
_________________________7. What is the route of absorption of cholesterol?
_________________________8. What are the two reduction products of cholesterol that is abundantly found
_________________________9. in the feces?
________________________10. In the biosynthesis of cholesterol, what substance is converted to
cholesterol?
________________________11. In the biosynthesis of cholesterol, acetyl CoA is converted to what
substance?
________________________12. The addition of cholesterol to phosphatidyl choline will produce this
substance and cholesterol esters.
II. Matching Type: Match column A (Reaction) to column B (Enzyme). Write the letter of your choice on
the space provided before each item.
Answer Column A: Reaction Column B: Enzyme

1. Oxidation of the alcohol group of hydroxyacyl CoA A. Acyl CoA DH
2. Cleavage of Acetyl CoA B. Lipase
3. Oxidation of Fatty Acyl CoA to form an α,β unsaturated C. Hydroxy-acyl CoA synthase
derivative
4. Addition of water to the trans alkene D. Enoyl CoA hydratase
5. Hydrolysis of triglyceride to fatty acid and glycerol E. Thiokinase
6. Enzyme catalyzing the formation of Acyl CoA esters F. Hydroxy-acyl CoA DH
7. Stimulates the entry of higher fatty acids in the G. Carnitine fatty acid transferase
mitochondria
8. Electron acceptor if the alcohol group of hydroxy-acyl H. α,β unsaturated derivative
CoA is oxidized synthetase
I. FAD
J. Thiolase
K. NAD
ASSIGNMENT:
Scoring Rubrik
4 3 2 1
unsuccessful
question but had
Page 325 of 334

answered each part only partial
accurately accuracy
correctly cited
slows readability
1. Name 5 diseases associated with lipid metabolism. Describe their biochemistry clearly.
2. Why do some alcoholics have high VLDL levels?
SUMMATIVE ASSESSMENT: TBA
Page 326 of 334

LABORATORY
INTRODUCTION
Urine is a clear, transparent fluid that normally has an amber color. The average amount of urine
excreted in 24 hours is between 5 to 8 cups or 40 and 60 ounces. Chemically, urine is mainly a watery solution
of salt and substances called urea and uric acid. Normally, it contains about 960 parts water to 40 parts solid
matter. Abnormally, it may contain sugar, albumin, bile pigments or abnormal quantities of one or another of its
normal components.
The analysis of urine or urinalysis is an integral part of a doctor’s routine patient examination because
of the two unique characteristics of a urine specimen which are: (a) that urine is readily available and easily
collected specimen; (b) that urine contains information about many of the body’s major metabolic functions,
and this information can be obtained by inexpensive laboratory tests. These characteristics of urine have
become the basis of modern urinalysis which includes not only the physical examination of urine but also the
chemical analysis and microscopic examination of the urinary sediment.
In general, urine consists of urea and other organic and inorganic chemicals dissolved in water.
Considerable variations in the concentrations of these substances can occur owing to the influence of factors
such as dietary intake, physical activity, body metabolism, endocrine functions, and even body position.
Increased amounts of these formed chemical substances and elements are often indicative of disease.
Activity No. 21
BODY FLUIDS: URINE
OBJECTIVES

1. describe correctly the physical properties of urine
2. perform precisely some tests to identify the physical properties of urine
3. trace accurately how urine is formed
MATERIALS
Test tubes Bunsen Burner Urine test strips

Test tube rack Vial for urine 3% Acetic acid
Test tube holder Benedict’s reagent 5% acetic acid
Test tube brush 0.5% Na nitroprusside Hydrogen peroxide
Pasteur pipette conc. NaOH Benzidine
PROCEDURES AND OBSERVATIONS:

Scoring rubric
For Procedures:
Page 327 of 334

For Rationalization
not clear
not clear
Using an appropriate container, collect at least 20 mL of urine using the midstream catch. Follow the
universal precautions when performing this activity. Personal protective equipment should be worn all the time.
Treat all specimens as a potential pathogen.
Physical Properties of Urine
1. Place 3 mL of urine in a clean and dry test tube and note the following
• Color: _________________________________
• Turbidity : ______________________________
• pH: ___________________________________
• specific gravity: __________________________
2. Test strips are available for pH and specific gravity. Ask your instructor on how to use it.
Watch: https://www.youtube.com/watch?v=g_4PiTezF6s
Chemical Properties of Urine
Test for Proteins: Heat and Acetic Acid Test

Watch: https://www.youtube.com/watch?v=vfua4bB1Btg
1. Place 3 to 5 ml of urine in test tube

2. Tilt the tube and boil the upper portion using the Bunsen burner
3. If turbidity develops, add 1 – 2 drops of 3% glacial acetic acid.
• Sometimes turbidity may be due to phosphate or carbonate precipitation. The glacial acetic acid
is used to clear this up. If he turbidity is due to proteins, it will remain even after the addition of
proteins
4. Reboil the upper portion of the specimen
5. If turbidity persists on the upper part of the test tube, then protein is present
6. Grade the turbidity as follows
• Negative : No cloudiness
• Trace: Barely visible cloudiness
• 1+: definite cloud with granular flocculation
• 2+: heavy and granular cloud without granular flocculation
• 3+ denssed cloud with marked flocculation
• 4+: thick curdy precipitation and coagulation
Observations and Interpretations:
Page 328 of 334
___________________________________________________________________
___________________________________________________________________
Test for reducing sugars: Benedict’s test
Watch: https://www.youtube.com/watch?v=-atHARq0JbQ
1. Pipette 1 mL of Benedict’s reagent in the test tube

2. Add 8 drops of your urine sample
3. Place in a boiling water bath for 5 – 10 minutes
4. Remove from the water bath after the stipulated time
5. interpret as follows
• Negative: No change in color i.e. blue
• Trace: Pale green with slightly cloudy
• 1+ : Definite cloudy green
• 2+ : Yellow to orange precipitate with supernatant fluid pale blue
• 3+ : Orange to red precipitate supernatant fluid pale blue
• 4+ : Brick red precipitate supernatant decolorized
Observations and Interpretations:
__________________________________________________________________________________
____________________________________________________
What reducing sugar is usually tested in urinalysis?
___________________________________________________________________
Test for ketone bodies: Legal’s test

Watch: https://www.youtube.com/watch?v=fe_KGUeTH60&list=TLPQMTUwNzIwMjDYXHuIaRzR7Q&index=2
1. Place 2 mL of urine in a test tube

2. Add 2 drops of 0.5% sodium nitroprusside solution
3. Note any color changes:
___________________________________________________________________
4. Add 5 drops of conc. NaOH solution
5. Note any color changes:
___________________________________________________________________
What ketone body is usually tested for in urine?
___________________________________________________________________
Test for blood: Benzidine test
1. Place a pinch of benzidine in a test tube

2. Add 2- 3 drops of 5% glacial acetic acid and mix well
3. Add 2 ml of hydrogen peroxide solution and let stand.
4. Transfer the supernatant to another test tube and add a few drops of urine
Observe for any color change:
___________________________________________________________________
Page 329 of 334

___________________________________________________________________
__________________________________________________________________________________
____________________________________________________

1. Give the clinical significance of the following terms
a. Proteinuria
b. Glucosuria
c. Hematuria
d. Ketonuria
2. Briefy describe how urine is formed
3. Complete the table below:
Changes in Unpreserved Urine

Analyte Changes Cause/s
Color
Clarity
Odor
pH
Glucose
Ketones
Bilirubin
Urobilinogen
Page 330 of 334
Nitrite
RBC, WBC & Casts
Bacteria
Page 331 of 334

EVALUATION OF THE COURSE
Dear Student,
As we finish one semester of learning, the authors would like you to answer the following questions in all
honesty. This will not form part of your grade but will help us improve this module.
Thank you so much for your cooperation.
1. What lesson or activity did I enjoy most? Why?

2. What is the most important lesson which I can apply in my daily life?
3. What are the new insights/discoveries that I learned?
4. What topic/s do I find least important?
5. What possible topics should have been included?
Page 332 of 334

REFERENCES:
Angeles, Chita Celeste C. and Panlasigui, Leonora N. (2004) Fundamentals in Nutrition and Biochemistry, Manila,
Philippines: NDAP.
Bernaldez, Alicia T. & Hessari, Cecilia B. (2001) Basic Biochemistry for Health and Food Science Courses: Outlines and
Notes. Manila, Philippines: UST Publishing House.
Caret, Robert L.; Denniston, Katherine J. & Topping, Joseph J. (2004). Principles and Applications of Inorganic, Organic
and Biochemistry (6th ed) Iowa, USA: Times Mirror Higher Education Group, Inc..
Hein, Morris, Best, Leo, Pattison, Scott & Arena, Susan (2004). Introduction to General, Organic and Biochemistry (8th
ed.). California: Brooks/Cole Publishing Co.
McKee, T. & Mckee, J. (2003) Biochemistry: The Molecular Basis, (3rd ed.) Philadelphia, USA: McGraw-Hill Book Co.
Stoker, H. (2010). General, organic, and biological chemistry. (5th ed). Australia : Brooks, Cole, Cengage Learning,
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BIOCHEMISTRY

FOR UNIVERSITY OF BAGUIO

A Self-regulated Learning Module 2

Welcome to the world of Biochemistry!

A Self-regulated Learning Module 3

A Self-regulated Learning Module 4

COURSE TITLE: BIOCHEMISTRY

REQUIREMENTS OF THE COURSE:

A Self-regulated Learning Module 5

A Self-regulated Learning Module 6

A Self-regulated Learning Module 7

Teresa N. Villanueva, RMT, MACT

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Lesson 1. Definition, History and Importance of Biochemistry

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1. Freidrich Wohler (1828)

- His experiment laid the foundation for modern biochemistry

2. Two Major breakthroughs in the history of Biochemistry

a. Discovery of enzymes and its roles

b. Identification of Nucleic Acid as information molecules

• Oswald Avery, Colin Macleod and Maclyn McCarty (1944)

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Lesson 2. Branches of Biochemistry

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1. Fischer Projection – linear representation 2. Haworth Projection

- Ring form (biochemical form

Nucleic acids are polymers composed of monomers called nucleotides. Nucleotides

Molecular Physiology is concerned about the functions of the different biological

Work within cells

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3. Synthetic work – a change in chemical bonds required to generate complex molecules

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2 types of metabolic pathways

2. Cyclic – series of reactions regenerates the first reactant

1. ENZYMES glucagon lactase lipase sucrase

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2. CATABOLISM – is the destructive phase of metabolism. It is a continuous process concerned

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Oxidation Reduction Reactions

ELECTRON ACCEPTORS OR DONORS USED BY DEHYDROGENASE ENZYMES

FAD – from double to single bond

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1. Synthesis of a polysaccharide from monosaccharides ____________________

Lesson 4. Processes Undergone by Biomolecules

PROCESSESS UNDERGONE BY BIOMOLECULES

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A. IDENTIFICATION. Given the following descriptions/ definitions, identify what is being

_______________________________ 1. This a biomolecule that contains the elements

_______________________________ 3. In the following reaction:

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Name: _____________________________ Date: _____________________________

At the end of the laboratory session, you should be able to

PROCEDURES AND OBSERVATIONS:

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TIME/SCHEDULE: Week No. 2

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8 CELL is an independent, simplest structural unit of life. It is the fundamental unit of

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ORGANELLES OF LIVING CELLS

Name: _ Date: _

Name: _ Date: _

Name: _ Date: _

Name: _ Date: _