SESSION 9:

ENZYMES
 Complex organic compound w/c has the ability of speeding chemical reactions w/out being themselves affected in the
process.
 In the absence of an enzyme, the reaction may hardly proceed at all, whereas in its presence the rate can be increased up to
10-fold.
 The study of enzymes is known as Enzymology.
 Functions as being biological catalyst (catalyze only reactions in living systems)
 1,000 enzyme/cell = each enzyme has its own specific reaction

A. PHYSICOCHEMICAL NATURE OF ENZYME:

1. All enzymes are protein in nature. Enzymes follow the physical and chemical nature of proteins.
a. simple protein
b. conjugated protein: prosthetic group (non-protein in nature)
: makes the enzyme active
: "active site”

2. Because enzymes are protein in nature, they are sensitive to any or all of the denaturating agents including changes
on the pH of the medium
3. They are heat labile
4. They are water soluble
5. Colloids that are soluble in water
6. Work best at temperatures between 35-40*C
7. Activity is dependent on the pH of the medium
8. Highly selective
 Follow the principle of lock and key theory

9. They contain 16% weight as nitrogen.

10. Have unique characteristics such as shape, specificity, and function.
11. Have an active site for target molecules called substrates.
12. Are much larger in size than their substrates.
13. Are not used up or permanently changed by the reaction.
14. Can be recycled, thus function in extremely low concentrations.
15. Can be regulated by feedback and genetic mechanisms.

B. CO-FACTORS
 Non-protein group in an enzyme (prosthetic group)
 Where a co-factor enzyme lacks catalytic activity w/o them
 Protein portion is now called APOENZYME. It is heat labile.
 These two portions combined together are called the holo-enzyme.

*Some cofactors are only transiently associated with a given enzyme molecule, so that they function as cosubstrates
2 CO-FACTORS
1. Co-enzyme - nonprotein organic compounds and many coenzymes are derived from vitamin precursors which are often
essential components of the organism's diet, thus giving rise to deficiency diseases when in inadequate supply

 Enzymes + other substances such as vitamins or organic and inorganic substances -

 Non-enzyme part of the Co-enzyme is termed as Co-factor
 Zymogens or Proenzymes - inactive form of the enzyme
 Eg pepsinogen

Examples:
a. FAD - Flavine Adenine Dinucleotide

b. NAD - Nicotinamide Adenine Dinucleotide } commonly found in vitamins

c. FMN- Flavine Mononucleotide

Selected Vitamins and Their Coenzymatic Functions

 Biotin: involved in carbon dioxide fixation reactions and fatty acid synthesis.
 Vitamin B12 (cyanocobalamin): Coenzyme involved in the transfer of methyl groups; active in amino acid metabolism.
 Vitamin E: Needed for cellular and macrocellular syntheses
 Vitamin K: Coenzyme used in electron transport (naphthaquinones and quinones)

Salient features of co-enzymes

 The protein part of the enzyme gives the necessary three dimensional infrastructure for chemical reaction; but the
group is transferred from or accepted by the co-enzyme
 The co-enzyme is essential for the biological activity of the enzyme
 Co-enzyme is a low molecular weight organic substance. It is heat stable
 Generally, the co-enzymes combine loosely with the enzyme molecules. The enzyme and co-enzyme can be separated
easily by dialysis
 Inside the body, when the reaction is completed, the coenzyme is released from the apo-enzyme, and can bind to
another enzyme molecule.
 One molecule of the co-enzyme is able to convert a large number of substrate molecules with the help of enzyme
 Most of the co-enzymes are derivatives of vitamin B complex substances.

2. Metal-ion activator - inorganic ions

Examples:

FI, Cr, Mn, Fe, Co, Cu,

*Metalloenzyme contains an apoenzyme and a metal ion cofactor

REACTIONS:

1 Co-enzyme + apoenzyme - holoenzyme

2. Metal ion activator + apoenzyme - holoenzyme

Active site: is the region of the enzyme that binds the substrate, to form an enzyme-substrate complex, and transforms it into
product. The active site is a three-dimensional entity, often a cleft or crevice on the surface of the protein, in which the
substrate is bound by multiple weak interactions.

Coenzymes Must Be Regenerated.

Coenzymes are chemically changed by the enzymatic reactions in which they participate. In order to compete the catalytic
cycle, the coenzyme must return to its original state. For a transiently bound coenzyme (cosubstrate), the regeneration
reaction may be catalyzed by a different enzyme as we have seen to be the case for NAD+. However, for a prosthetic group,
regeneration occurs as part of the enzyme reaction sequence

NOMENCLATURE OF ENZYME:

1. Add - ase to the substrate eg. carbohydrase, lipase, protease

2. Add - ase to the reaction catalyst eg. oxidase, reductase, hydrolase

Example:

Lactase: the substrate catalyzed is lactose

Maltase: the substrate catalyzed is maltose

International Union of Biochemistry and Molecular Biology (IUBMB) in 1964, (modified in 1972 and 1978), suggested the
IUBMB system of nomenclature of enzymes. As per this system, the name starts with EC (enzyme class) followed by 4 digits.

 First digit represents the class

 Second digit stands for the subclass
 Third digit is the sub-sub class or subgroup
 Fourth digit gives the number of the particular enzyme in the list

Examples:

EC 1.1.1.2 (Alcohol Dehydrogenase)

SIX MAJOR CLASSIFICATIOPOF ENZYMES

CLASS NAME TYPE OF REACTION CATALYZED EXAMPLES

1 Oxidoreductases  Catalyzing oxidoreduction between two substances
2 Transferases
3 Hydrolases
Dehydrogenase, Oxidase
Lactate dehydrogenase
(NAD); Glucose-6-
phosphate
dehydrogenase (NADP);
Succinate
dehydrogenase (FAD):
dioxygenases.
 Catalyzing a transfer of group, other than hydrogen, Aminotransaminase,
between a pair of substrates. Hexokinase
 Catalyzing hydrolysis of ester, ether, peptide, Acetylcholine sterases,
glycosyl, acid-anhydride, C-C, C-halide or P-N bonds Digestive enzymes such
 Cleave bond and add water/Hydrolysis reaction as trypsin

4 Lyases  Catalyzing removal of groups from substrates by Decarboxylase, Aldolase

mechanisms other than hydrolysis, leaving double
bonds.
 Cleave without adding water
 5 Isomerases  Catalyzing interconversion of optic, geometric or Phosphohexoisomerase,
positional isomers. Fumarase, Triose
 Intramolecular transfers. phosphate isomerase.
 Transfer of groups within a molecule
6 Ligases  Catalyzing linkage of 2 compounds coupled to the Citric acid synthetase,
breaking of a pyrophosphate bond in ATP or a Acetyl COA carboxylase:
similar compound Glutamine synthetase;
 ATP dependent condensation of two molecules/ PRPP synthetase.
Joining of two molecules
 Ligase is also known as synthetase
 Difference between synthetase and synthase

Synthetases are ATP-dependent enzymes catalyzing biosynthetic reactions; they belong to Ligases (class 6). Examples are
Carbamoyl phosphate synthetase; Arginino succinate synthetase; PRPP synthetase and Glutamine synthetase.

Synthases are enzymes catalyzing biosynthetic reactions, but they do not require ATP directly, they belong to classes other
than Ligases. Examples are Glycogen synthase and ALA synthase.

CLASSIFICATION OF ENZYMES ACCORDING TO CHEMICAL REACTION CATALYZED:

1. ADDITION OF WATER MOLECULES

A. HYDROLASE

a) Carbohydrase - aid in the hydrolysis of carbohydrates

 alpha amylase - starch & glycogen to dextrin

 beta amylase - starch & glycogen to dextrin & maltose
 lactase - lactose to glucose and galactose
 maltase - maltose to glucose
 sucrase - sucrose to glucose and fructose

b) Esterases - aid in the hydrolysis of esters

 lipases - glycerides to fatty acids and glycerol

 phosphatase - organic phosphates to phosphoric acid
 choline esterase/cholinesterase - acetyl choline to acetic acid and choline

c) Nucleases - aid in the hydrolysis of nucleic acids

 polynucleotidases - nucleic acid to nucleotides

 nucleotidases - nucleotides to nucleosides and phosphoric acid
 nucleosidase - nucleosides to sugar and purine or pyrimidine

d) Amidases - aid in the hydrolysis of amides

 ureases - urea to ammonia and CO2

 arginase-arginine to ornithine and urea

e) Proteases - aid in the hydrolysis of proteins

 pepsin - proteins to proteoses and peptones
 trypsin - cleaves the polypeptide chain from the carboxyl end of arginine and lysine
 chymotrypsin - cleaves the polypeptide chain from the carboxyl end of phenylalanine, tyrosine and tryptophan
 rennin - casein to paracasein
 cathepsin - proteins to proteoses and peptones
 bromelin/bromelain - proteins to proteoses and peptones

f) Peptidases - hydrolyze peptides to simple peptides and amino acids

 aminopolypeptidases - sequentially cleave the polypeptide chain rom the amino terminal
 carboxypolypeptidases - sequentially cleave the polypeptide chain from the carboxyl terminal
 prolinase - polypeptides containing praline to simpler peptides and praline
 dipeptidases - dipeptides to amino acids
II. TRANSFER OF ELECTRONS

A: OXIDASES - are enzymes which catalyze the removal of hydrogen from a substrate and pass it directly to oxygen

a) Dehydrogenases - activate H atoms of organic compounds

b) Catalase - acts on hydrogen peroxide to give water and oxygen

c) Peroxidases - act on organic peroxides giving nascent oxygen

d) Tyrosinase - acts on tyrosine

e) Ascorbic acid oxidase - acts on ascorbic acid

III- SPLITTING OR FORMING A C-C BOND

A. DESMOLASES - catalyze the linkage not broken by water, splitting or forming a C-C bond

a) Decarboxylases

i. carboxylase - alpha keto acids to CO2 and aldehydes

ii. carbonic and anhydrase - carbonic acid to water and carbon dioxide

IV. TRANSFER OF A RADICAL

A. TRANSAMINASES - catalyze the transfer of amino groups from amino acids to ketoacids and thus promote the formation of
new amino acids

A. alanine transaminase - catalyzes the transfer of an alpha-amino group from an amino acid to pyruvic acid to produce
alpha-keto acid and alanine

B. glutamate transaminase - catalyzes the transfer of an alpha-amino group from an amino acid to alpha-ketoglutyaric acid,
to produce an alpha-keto and glutamic acid

SESSION 10:
SOME ENZYMES WITH THEIR SOURCES AND USES (Commercial, Pharmaceutical
and Medicinal):
1. PROTEOLYTIC ENZYMES

a. Pepsin

 obtained from the stomach of Sus scrofa, Fam. Suidae

 Use: assist in gastric digestion
 Found in gastric juice
 In neutral or alkaline environment, it is inactive
 Converts proteins to proteases and peptones
 Proenzyme: Pepsinogen which is activated by HCI
 Protein digestion starts in the stomach
 Digestive aid in pre-cooked foods

b. Alcalase
 additive to remove protein stains

c. Bromelains
 mixture of protein-digesting & milk clotting enzymes from the juice of Ananas comosus, Fam. Bromeliaceae
 Meat tenderizer
 Use: Adjunctive therapy to reduce inflammation & edema
 Accelerate tissue repair, especially following episiotomy
d. Papain

 Obtained from the latex of Carica papaya, Caricaceae

 Use - Relieve symptoms of episiotomy - Cleaning solutions of soft contact lenses
 Meat/Beef tenderizer
 As skin whitener (Papain, natural enzyme that promotes skin renewal and cell turnover. It helps in exfoliating the skin
to reveal newer cells, and has restorative properties that soften the skin.)
 Stabilizer for chill-proof beer
 Chymopapain
 Nonpyrogenic proteolytic enzyme obtained from the latex of Carica papaya, Fam. Caricaceae
 It is a sulfhydryl enzyme similar to papain with respect to substrate specificities but differing in electrophoretic,
mobility, stability and solubility
 Employed in the treatment of herniated lumbar intervertebral discs, chymopapain is injected into the nucleus
pulposus to hydrolyze the noncollagenous polypeptides or proteins that maintain the tertiary structure of the
chondromucoprotein
 Relieves the comprehensive symptoms of lower back pain by lessening osmotic activity and thereby decreasing
fluid absorption and reducing intradiscal pressure

e. Trypsin

 Converts proteases and peptones to polypeptides and amino acids

 Secreted by the pancreas - Released in the intestines - Active in an alkaline environment
 For wound debridement
 Chymotrypsin
 Crystallized from an extract of the pancreatic gland of Ox (Bos Taurus, Fam Bovidae):
 For ophthalmic solution
 Administered in solution to the posterior chamber of the eye, under the iris, to achieve zonal lysis

f. Sutilains

 from Bacillus subtilis

 Use: Wound Debridement

g. Rennin

 Coagulating enzyme that is present in the mucous membrane of the stomach of animals
 It curdles the soluble casein in milk
 For cheesemaking

h. Erepsin
 Found in the intestinal juice
 Converts proteoses and peptones into amino acids

i. Streptokinase
 It is a purified bacterial protein elaborated by group C B-hemolytic streptococci
 Supplied as lyophilized powder
 Acts to convert plasminogen to plasmin (degrades not only fibrin clots but also fibrinogen and other plasma proteins)
 Use: treatment of pulmonary embolism, deep vein thrombosis, arterial thrombosis and embolism, arteriovenous
cannula acclusion and coronary artery thrombosis

j Urokinase
 Isolated from human urine or obtained from human kidney cells by tissue culture techniques
 Converts plasminogen to enzyme plasmin
 Use: treatment of pulmonary embolism, coronary artery thrombosis, and restoring the patency of intravenous
catheters

k. Fibrinolysin
 In the blood serum as a protease and in plasma as the inactive precursor, profibrinolysin (or plasminogen)
 It is prepared commercially by activating a human blood plasma fraction with streptokinase
 Use: treatment of blood clots within the cardiovascular system, exclusive of thrombi of the coronary and cerebral
ateries
2. OXIDIZING ENZYMES

i. Peroxidase

 Widely distributed in plants

 Bring about the oxidation reactions that cause the discoloration of bruised fruits

ii. Thrombin

 Converts the fibrinogen of the circulating blood into the insoluble fibrin of the blood clot
 Also known as Coagulation factor II, plays a physiological role in regulating hemostasis and maintaining blood
coagulation. Once converted from prothrombin, thrombin converts fibrinogen to fibrin, which, in combination with
platelets from the blood, forms a clot

3. CARBOHYDRASEIAMYLOLYTIC ENZYMES

a. Amylase

 found in the salivary glands

 a.k.a. salivary diastase or ptyalin
 digestive aid in pre-cooked foods

b. Diastase

 A yellowish white, amorphous powder obtained from an infusion of malt

 Can convert 50 times its weight of potato starch into sugars

c. Amylopsin

 found in the digestive tract of animals

 sometimes called "animal diastase"

d. Invertase or sucrase

 found in yeast and in the intestinal juice

 aids in the hydrolysis of sucrose into glucose and fructose
 prevention of granulation of sugar in soft-centered candies

e. Zymase

 Fermenting enzyme causing the conversion of monosaccharides (glucose, fructose) into alcohol and CO2

f. Emulsin

 Found in almonds
 Causes the hydrolysis of B-glucosides; thus, amygdalin is hydrolyzed into glucose, benzaldehyde and hydrogen cyanide

g. Myrosin

 Found in white and black mustard

 Hydrolyzes sinalbin, sinigrin and other glycosides

h. Amyloglucosidase

 Production of dextrose from starch

i. Cellulase
 Preparation of coffee liquid concentrate

j. Lactase
 Prevention of lactose crystals in ice cream

k. Pectinase
 Clarification of wines and juices
I. Hyaluronidase
 Mucolytic enzyme capable of depolymerizing and catalyzing hyaluronic acid and similar hexosamine-containing
polysaccharides
 Spreading and a diffusing factor/agent
 It occurs in human testes, in various bacterial cultures as a metabolic product, in heads of leeches and in snake venoms
 Promotes diffusion and hastens absorption of subcutaneous infusions

4. ESTERASES

a. Lipase

 lipolytic enzyme widely distributed in the animal and vegetable kingdom

 found in the pancreatic juice of animals and in oily seeds
 hydrolyzes fats into glycerin and fatty acids
 flavor production in cheese

b. Pectase

 Splits pectin into pectic acid and methyl alcohol

c. Steapsin

 Lipolytic enzyme capable of digesting dietary fat

d. Urease

 Obtained from soybeans

 Used as a laboratory reagent for converting urea to ammonia

5. OTHERS

a. Collagenase

 An enzyme preparation obtained from fermentive cultures of Clostridium histolyticum

 Cleaves collagen and is used topically to debride dermal ulcers and severely burned areas
 Care should be exercised to avoid heavy metal inactivation of the enzyme; Burow's solution can be used to stop the
enzyme's action if the risk of bacteremia develops

b. L-Asparaginase

 An enzyme obtained from cultures of certain stains of Escherichia coli

 Induces hematologic and clinical remissions of short duration in a significant percentage of children with acute
leukemia
 The antitumor effect may be attributed to degradation by the enzyme of circulating L-asparagine, which results in the
death of cells that require exogenous sources of this amino acid for survival
 Adverse reactions: allergic reaction and fatal anaphylaxis

c. Lipoxygenase

 Bread – whitening
SESSION 11:
MODE OF ENZYME ACTION:

1. The surface of the substrate contacts a specific region of the surface of the enzyme molecule called active sites

2. A temporary intermediate compound forms, called an enzyme-substrate complex

3. The substrate is then transformed into products.

4. The products are released.

5. The enzyme is recovered unchanged

 Activation Energy: the minimum amount of energy that is required to activate atoms or molecules to a condition in
which they can undergo chemical transformation or physical transport.

THEORIES EXPLAINING THE MORE OF ENZYME ACTION

I. FISCHER'S LOCK AND KEY THEORY

 Enzyme - substrate (F-5) complex theory

1. The active site of the enzyme and substrate have complementary structures, hence they fit together as a key fits a lock

2. While they are bounded in the enzyme-substrate complex, catalytic reaction occurs

3. The products of the reaction leave the surface of the enzyme & combine with another molecule of the substrate

Examples:

General Reaction:

1. E + S == E-S complex

2. ES complex - product + enzyme

Eg.

1. sucrase + sucrose == sucrase - sucrose complex

2. sucrase- sucrose complex == glucose + fructose + sucrose Lock and key hypothesis
 II. KOSHLANDSHJCED FIT THEORY
 In this model the substrate still needs to fit into the enzyme like a key, but instead of simply fitting into the "keyhole,"
some type of modification is induced in the substrate, enzyme, or both. The modification begins the process of the
reaction.
 At first, substrate binds to a specific part of the enzyme. This leads to more secondary binding and conformational
changes. The substrate induces conformational changes in the enzyme, such that precise orientation of catalytic groups
is effected

FACTORS THAT INFLUENCE EZNZYME ACTION:

1. Concentration of substrate

 | concentration of substrate = | enzyme action

2. Concentration of enzyme

 | concentration of enzyme = | enzyme action

3. Temperature

 Optimum temperature of enzyme in the body = 37°C

 With each 10°C rise in temperature usually doubles or triples reaction rate
 Rapid inactivation of the enzyme occurs at temperatures much above body temperature
 Low temperatures slow down the action of enzyme but do not destroy them

4. pH
 each enzyme has its specific optimum pH at which it exerts its maximum activity
 it loses activity rapidly on either side of this opt.

Eg. Pepsin - 1.5 to 2.2

Lactase - 5.7

Trypsin - 7.8

5. Presence of accelerators

 Activate the enzyme

Eg. Metallic ions such as Ferrous, Ferric, Co. Mn, Mg, Mo

6. Presence of inhibitors

 Have the ability to combine with enzyme in a reversible and irreversible reaction and hence block enzyme catalysis

Eg. Antibiotics - streptomycin

Antienzymes - antitrypsin from soybeans

Antimetabolites - sulfanilamide

Others (Poisons/Drugs) – formaldehyde, chloroform, CCI4, cyanides, heavy metals like Ag and Hg
7. Environmental hazards

Eg. 1. Hg (mercury)

a. Hg from phenyl mercuric acetate

 Fungicide to prevent cellulose from rotting

b Methyl mercuric acetate

 Prevent rotting of seed grains

2. Cd (cadmium)

 Derived from cigarette smoke

 1 pack = 15 mcg

3. Pb (lead)

 Present in gasoline
 Yellow-painted pencils

ENICAL ENZYNOLOGY:

 Enzymes are commonly used in medicine as:

1. Analytical tools or reagents in measuring quantities of various constituents in biologic fluids

Eg. Urease, alcohol dehydrogenase

Enzyme Used for Testing

Urease Urea
Uricase Uric acid
Glucose Oxidase Glucose
Cholesterol oxidase Cholesterol
Lipase Triglycerides
Reverse Transcriptase Polymerase Chain Reaction

2. As index for pathology of diseases:

Eg of some serum enzymes:

 a-amylase in pancreatitis - normal serum level is elevated

 Lipase - 1 in acute pancreatitis, perforated duodenal and peptic ulcers and intestinal obstruction
 Creatinine Phosphokinase (CPK) - | in blood after MI
 It is a sensitive indicator in the early stages of myocardial ischemia.
 Three CK isoenzymes
 MM(CK3) - found in skeletal and heart muscle
 MM(CK2) - found in heart muscle
 BB(CK1) - found in the brain
 Alkaline Phosphatase - | in rickets, obstructive jaundice, cirrhosis and cholestasis
 Acid Phosphatase - | in carcinoma of the prostate or prostate cancer
 Lactase dehydrogenase (LDH) - | in viral heap heart and skeletal muscle disease
 Aspartate transaminase (AST) - | in myocardial infarction
 Alanine transaminase (ALT) - | in liver disease (liver cell damage)
 Glutamyl transferase - | in liver disease, biliary obstruction (gallstone formation), and alcoholism
 5' Nucleotidase - Moderately increased in hepatitis and highly elevated in biliary obstruction.
SESSION 12:
NUCLEIC ACIDS
 Consist of long-chained nucleotides (polynucleotides) combined with another thru a PHOSPHATE DIESTER LINKAGEE
 Nucleotides linked by 3’, 5’phosphodiester bonds
 Sequence always specified as 5’-3’
 Macromolecules found in all cells, which participate in the storage, transmission and translation of genetic information

BIOLOGICAL SIGNIFICANCE:

1. Responsible for storage and transmission of genetic information code

2. For the precise synthesis of protein characteristics of individual cell

PROPERTIES OF NUCLEIC ACIDS:

1. Insoluble in alcohol
2. Slightly soluble in cold water but readily dissolved in hot water and dilute alkalies, forming alkali salts
3. Precipitated by HCI and by excess of acetic acid

2 TYPES OF NUCLEIC ACIDS:

1. RNA - Ribonucleic acid (the nucleic acid responsible for using the genetic information encoded in DNA to produce the
thousands of proteins found in living organisms.)

2. DNA-Deoxyribonucleic acid (the nucleic acid that stores genetic information)

DNA- Deoxyribonucleic Acid

 Molecules of heredity
 Made up of mononucleosides d-AMP; d-GMP; d-TMP linked in various sequence with 3' - 5' phosphate diester linkage
 Variable: base sequence
 Backbone: P - sugar - P - sugar
 DNA + histone - chromatin complex
 Base pairing rule: A = T

G=C

 Found in the nucleus of cells as a double-stranded, highly twisted polymer

 Double-coiled chain of nucleotides
 Resemble a ladder that has been twisted into a helix
 Anti-parallel - one side runs fr. 5'-3' while the other side runs Fr. 3-5' (opposite)

DNA-ANTIPARALLEL
DNA TYPES

There are three different DNA types:

 A-DNA: It is a right-handed double helix similar to the B-DNA form. Dehydrated DNA takes an A form that protects the DNA
during extreme condition such as desiccation, protein binding also removes the solvent from DNA and the DNA takes an A
form.
 B-DNA: This is the most common DNA conformation and is a right-handed helix. Majority of DNA has a B type
conformation under normal physiological conditions.
 Z-DNA: Z-DNA is a left-handed DNA where the double helix winds to the left in a zig-zag pattern. It was discovered by
Andres Wang and Alexander Rich. It is found ahead of the start site of a gene and hence is believed to play some role in the
gene regulation.

Different forms of DNA

WHO DISCOVERED DNA?

 DNA was first recognized and identified by the Swiss biologist, Johannes Friedrich Miescher in 1869 during his research on
white blood cells.
 The double helix structure of a DNA molecule was later discovered through the experimental data by James Watson and
Francis Crick. Finally, it was proved that DNA is responsible for storing the genetic information of a human being.

CHARGAFF'S RULE

 Erwin Chargaff, a biochemist, discovered that the number of nitrogenous bases in the DNA was present in equal
quantities. The amount of A is equal to T, whereas the amount of C is equal to G.

A=T; C=G

 In other words, the DNA of any cell from any organism should have a 1:1 ratio of purine and pyrimidine bases.

FUNCTIONS:

1 To make exact copies of itself in the process of replication

2. To transfer the genetic information to m-RNA in the process of transcription

 GENOME - DNA that constitutes the total genetic information content of an organism, the segments of the genome that can
be translated is called GENE
 Introns -DNA segment that do not convey code for genetic information
 Exons - DNA segments that convey genetic information
 ALPHA - HELIX - WATSON -CRICK MODEL

.
RNA- Ribonucleic Acid
 Single chain of nucleotides
 Made up of major mononucleotides: AMP, GMP, UMP, CMP
 Variable: base sequence
 Backbone: P- sugar - P- sugar

3 FORMS OF RNA:

1. Messenger RNA (MRNA)

 Made in the nucleus of the cell as a component to a DNA strand
 Pairing nile: UA; GC
FUNCTION:
 Serves as cytoplasmic messenger of genes and carrier of genetic information for protein synthesis
2. Ribosomal RNA (TRNA)
 Comprise 70-80% of the total cell RNA (most abundant)
FUNCTION:
 bind the m-RNA and a specific enzyme for protein synthesis
 It is used as a structural component of the ribosome.

3. transfer RNA (tRNA)

 10-15% of the total RNA content of the cell (second most abundant)
FUNCTION:
 Carry specific amino acids to the ribosomes and decodes the genetic information in mRNA in terms of proper amino
acid sequence

COMPOSITION OF NUCLEIC ACIDS:

 Nucleic acids are substances with high molecular weight ranging from 1286 to 3,000,000
 Made up of carbon, hydrogen, oxygen, nitrogen and phosphorous
 Nitrogen is from 15-16%, while phosphorous is from 9-10%

COMPOUND DNA RNA

1. Nitrogenous base
1.a. Pyrimidine base Cytosine/Thymine Cytosine/Uracil
1.b. Purine base Adenine/Guanine Adenine/Guanine
2. Sugar Deoxyribose Ribose
3. Acid H3PO4 H3PO4

1. N-base:

1.a. Pyrimidine

 Isolated from the hydrolysates of nucleic acid

 Pyrimidine bases have only one ring.
 Pyrimidine such as:
 Vitamin B1 (Thiamine) - plays an important role in metabolic processes
 Alloxan (2, 4, 5, 6-tetraoxypyrimidine) produces experimental diabetes and has helped in the study of the
nature of diabetes mellitus
 Thiouracil/Propylthiouracil - used as antithyroid drugs
1.b. Purine

 Made up of six membered pyrimidine ring and a five membered imidazole ring (two rings)
 Important purines occurring in nature which are metabolic products in animals
 Uric acid - weak acidic property
 Interacts with sodium potassium forming mono and disodium or potassium urates
 With its salts are sparingly soluble in water, hence the gradual precipitation of urates from the urine
upon standing
 Hypoxanthine
 Xanthine
 Caffeine (1, 3, 7-trimethylxanthine) from tea and coffee
 Theobromine (3,7-dimethylxanthine) from tea and cocoa

2. SUGAR

3. ACID PHOSPHORIC ACID

HYDROLYSIS OF NUCLEIC ACID:

NUCLEOSIDES

 Nitrogenous base + sugar

 Are derivatives of purines and pyrimidines that have a sugar linked to a ring nitrogen of a purine or pyrimidine
 Formed between C-1 of the sugar and N-1 of pyrimidine base or N-9 of Purine base; water is eliminated in the process

COMMON NUCLEOSIDES OF RNA:

A. Pyrimidine:
1 Cytidine = (Cytosine + Ribose)
2. Uridine = (Uracil + Ribose)

B. Purine:
1. Adenosine = (Adenine + Ribose)
2. Guanosine = (Guanine + Ribose)

COMMON NUCLEOSIDES OF DNA:

A. Pyrimidine:
1 Deoxycytidine = (Cytosine + Deoxyribose)
2. Deoxyuridine = (Uracil + Deoxyribose)
B. Purine:
1. Deoxyadenosine = (Adenine + Deoxyribose)
2. Deoxyguanosine = (Guanine - Deoxyribose)

NUCLEOTIDES
 Functional subunits of nucleic acids
 Are formed when 1 or more phosphate groups is attached to the 5'carbon of a nucleoside
 Nucleoside + Phosphoric acid
 The sugar component of nucleotide is in D-configuration.

NAMING OF NUCLEOTIDES:

A. As an ordinary compound B. As an acid

1. Name the nucleoside used 1. Change the -ine ending of the nucleoside to-ylic
2. Indicate the point of attachment (may be omitted) 2. Affix the term “acid"
3. Attach the word "monophosphate"

Eg cytidine - 5’ - monophosphate or cytidine Eg. cytidilic acid

monophosphate

COMMON NUCLEOTIDES OF RNA:

1. Cytidine monophosphate or cytidylic acid
2. Uridine monophosphate or uridylic acid
3. Adenosine monophosphate or adenylic acid
4. Guanosine monophosphate or guanylic acid

COMMON NUCLEOTIDES OF DNA:

1. Deoxycytidine monophosphate or Deoxycytidylic acid
2. Deoxyuridine monophosphate or Deoxyuridylic acid
3. Deoxyadenosine monophosphate or Deoxyadenylic acid
4. Deoxyguanosine monophosphate or Deoxyguanylic acid
SESSION 13:
THE CENTRAL DOGMA
 Is a series of theories of the transmission of hereditary information and protein synthesis.
 In Molecular Biology-this is the concept that regulate cellular activities through REGULATION OF PROTEIN

SYNTHESIS
 By Francis Crick in 1956, when he first proposed that the sequence of nucleotides in DNA determines the sequence of
amino acids in a protein
 Gene Expression: the process by which the genetic code - the nucleotide sequence - of a gene is used to direct protein
synthesis and produce the structures of the cell. Genes that code for amino acid sequences are known as

 DNA when it replicates will transfer genetic information from parent to offspring
 Replication DNA to DNA
o is a copying process by which DNA is applied to the new cells formed by cell division
o This genetic information is transferred to mRNA through Transcription
 Transcription DNA to RNA
o This process involves the transfer of genetic info from a DNA strand thru base pairing to form
complementary ribonucleotides, an RNA chain
 Translation RNA to PROTEINS
o This information is translated from nitrogenous base sequence to an amino acid sequence by tRNA as
presented to it by the ribosomes forming proteins RNA to PROTEIN

3 REACTIONS:
1. Replication - parent DNA to daughter DNA
2. Transcription - DNA to messenger RNA
3. Translation - mRNA to CHỌN's (reading is from 5 to 3')
 SEME-CONSERVATIVE REPLICATION-only 1 strand will undergo transcription

DNA TRANSCRIPTION
 Transcription is the synthesis of a complementary strand of RNA from a DNA template which takes place inside the
nucleus.
During transcription, a strand of mRNA is synthesized using a specific portion of the cell's DNA as a template. In other words,
the genetic information stored in the sequence of nucleobases of DNA is rewritten so that the same information appears in the
base sequence of mRNA.

As in DNA replication, a guanine (G) in the DNA template dictates a cytosine (C) in the mRNA being made and a C in the DNA
template dictates a G in the mRNA. Likewise, a thymine (T) in the DNA template dictates an adenine (A) in the mRNA. However,
an adenine in the DNA template dictates an uracil (U) in the mRNA, because RNA contains uracil instead of thymine. (Uracil has
a chemical structure slightly different from thymine, but it base-pairs in the same way.) If, for example, the template portion of
DNA has the base sequence 3'-ATGCAT, the newly synthesized mRNA strand will have the complementary base sequence 5'-
UACGUA.
The process of transcription requires both an enzyme called RNA polymerase and a supply of RNA nucleotides. Transcription
begins when RNA polymerase binds to the DNA at a site called the promoter. Only one of the two DNA strands serves as the
template for RNA synthesis for a given gene. Like DNA, RNA is synthesized in the 5' to 3' direction. RNA synthesis continues
until RNA polymerase reaches a site on the DNA called the terminator.

INHIBITORS OF RNA SYNTHESIS

1. Actinomycin D and Mitomycin: intercalate with DNA strands, thus blocking transcription. They are used as anticancer drugs.
2. Rifampicin is widely used in the treatment of tuberculosis and leprosy.
3. Quinolones and Fluoroquinolones: inhibit DNA gyrase (prokaryotic topoisomerase II), preventing DNA replication and
transcription.

3 STEPS OF TRANSCRIPTION
1. Initiation is the beginning of transcription. It occurs when the enzyme RNA polymerase binds to a region of a gene
called the promoter. This signals the DNA to unwind so the enzyme can "read" the bases in one of the DNA strands.
The enzyme is now ready to make a strand of mRNA with a complementary sequence of bases.

2. Elongation is the addition of nucleotides to the mRNA strand. RNA polymerase reads the unwound DNA strand
and builds the mRNA molecule, using complementary base pairs. There is a brief time during this process when the
newly formed RNA is bound to the unwound DNA. During this process, an adenine (A) in the DNA binds to a uracil
(U) in the RNA.

3. Termination is the ending of transcription, and occurs when RNA polymerase crosses a stop (termination)
sequence in the gene. The mRNA strand is complete, and it detaches from DNA.

DNA TRANSLATION
During translation, codons of an mRNA are "read" sequentially; and, in response to each codon, the appropriate amino acid is
assembled into a growing chain. The site of translation is the ribosome, and transfer RNA (TRNA) molecules both recognize the
specific codons and transport the required amino acids.

Each tRNA molecule has an anticodon, a sequence of three bases that is complementary to a codon. In this way, tRNA
molecules can base-pair with its associated codon. Each tRNA can also carry on its other end the amino acid encoded by the
codon that the tRNA recognizes. The functions of the ribosome are to direct the orderly binding of tRNAs to codons and to
assemble the amino acids brought there into a chain, ultimately producing a protein.

The two ribosomal subunits, a tRNA with the anticodon UAC, and the mRNA molecule to be translated, along with several
additional protein factors, all assemble. This sets up the start codon AUG) in the proper position to allow translation to begin.
After the ribosome joins the first two amino acids with peptide bond the first tRNA molecule leaves the ribosome. The
ribosome then moves along the mRNA to the next codon. As the proper amino acids are brought into line one by one, peptide
bonds are formed between them, and a polypeptide chain results.

Translation ends when one of the three nonsense codons in the mRNA is reached. The ribosome then comes apart into its two
subunits, and the mRNA and newly synthesized polypeptide chain are released. The ribosome, the mRNA, and the tRNAs are
then available to be used again.

INHIBITORS OF PROTEIN SYNTHESIS

1. Reversible inhibitors in Bacteria (Bacteriostatic Agents)
a. Tetracycline
b. Chloramphenicol
c. Erythromycin
d. Clindamycin
2. Irreversible inhibitors in Bacteria (Bactericidal Agents)
a. Streptomycin

IMPORTANT ENZYMES IN CENTRAL DOGMA

DNA Gyrase Relaxes supercoiling ahead of the replication fork
DNA Ligase Makes covalent bonds to join DNA strands; Okazaki fragments, and new segments in
excision repair
DNA Polymerases Synthesize DNA: proofread and facilitate repair of DNA
Helicase Unwinds double-stranded DNA
Primase An RNA polymerase that makes RNA primers from a DNA template
RNA Polymerase Copies RNA from a DNA template
MUTATION

 Any chemical or physical change that alters the sequence of bases in the DNA molecule.
 Any alteration in the protein as a result of a change in all cell structure, because when the genetic information in the
DNA is altered, the message transcribed into RNA will also be altered
 MUTAGENS - substance that causes mutation either physical or chemical form
TYPES OF MUTATIONS

SILENT MUTATION If abase substitution occurs in the third position of the codon there is a good chance that a
synonymous codon will be generated. Thus the amino acid sequence encoded by the gene is
not changed and the mutation is said to be silent. "new codon specifies same amino acid"
MISSENSE MUTATION This type of mutation is a change in one DNA base pair that results in the substitution of one
amino acid for another in the protein made by a gene. "New codon specifies different amino
acid"
NONSENSE MUTATION A nonsense mutation is also a change in one DNA base pair. Instead of substituting one amino
acid for another, however, the altered DNA sequence prematurely signals the cell to stop
building a protein. This type of mutation results in a shortened protein that may function
improperly or not at all. "new codon is stop codon"
FRAMESHIFT This type of mutation occurs when the addition or loss of DNA bases changes a gene's reading
MUTATION frame A reading frame consists of groups of 3 bases that each code for one amino acid. A
frameshift mutation shifts the grouping of these bases and changes the code for amino acids.
The resulting protein is usually nonfunctional. "Insertions deletions and duplications can all
be frameshift mutations.”

GENETIC CODE

 Set of rules which give a relationship between the nitrogenous bases and the amino acids in a polypeptide chain.

4. FEATURES:

1. The genetic code is degenerate - most amino acids, assume > 1 codon; however there is no one codon that specifies for > 1
amino acid
2. coding ratio - 3 bases (letters) coding for 1 amino acid
3. There are no punctuation marks to signify the start or the end of the chain
4. 3 codon will indicate the termination of the chain
Eg. UAG, UAA, UGA

BASIC PRINCIPLES OF HEREDITY

Terminologies

 Alleles: Genes that may replace one another at the same locus. responsible for alternate or contrasting characters.
 Homozygous: When both alleles carry the same defect.
 Heterozygous: When one allele is normal, and the counterpart is defective;
 Phenotype: The observed character expressed by the gene.
 Genotype represents the set pattern of genes present in the cell.
AUTOSOMAL DOMINANT INHERITANCE  Examples of diseases with autosomal dominant
inheritance are chondrodystrophy (dwarfism) and
 only one copy of a disease allele is necessary for an Huntington disease.
individual to be susceptible to expressing the
phenotype.
 Affected men and women transmit the abnormality
to their children. When an affected heterozygote
parent (Dd) has a normal spouse (dd), half of the
progeny will have the disease.
 With each pregnancy, there is a one in two (50%)
chance the offspring will inherit the disease allele.
 Male-to-male transmission can be observed.

AUTOSOMAL RECESSIVE INHERITANCE

 In autosomal recessive inheritance, two copies of a  Examples of diseases with autosomal recessive
disease allele are required for an individual to be inheritance include phenylketonuria, albinism,
susceptible to expressing the phenotype. galactosemia, sickle cell anemia and cystic fibrosis.
 Typically, the parents of an affected individual are
not affected but are gene carriers.
 When both father and mother are carriers, one-
quarter of siblings express the disease (both alleles
abnormal) another one-quarter of siblings are
normal, and half of the children are carriers.
 As with autosomal dominant inheritance, the
proportion of affected males should be equal to the
proportion of affected females in a given
population.

SEX-LINKED (X-LINKED) RECESSIVE INHERITANCE

 In the autosomal conditions, the disease occurs in in males who are hemizygous (XY) for the
both sexes with equal frequency. But in sex-linked condition, but not in-females who may be
conditions, X-chromosome carries the abnormal heterozygous (XX)
gene.  Hemophilia, glucose-6-phosphate dehydrogenase
 When a normal male carries a carrier female deficiency, pseudohypertrophic muscular
(unaffected parent), the children can be affected dystrophy (Duchenne type), and red-green color
male (25%), female carrier (25%), normal male blindness are examples of sex-linked recessive
(25%) and normal female (25%). inheritance.
 All male children of an affected male and normal
female will be normal; but all female children will
be carriers since they inherit the abnormal X from
their father
 There is no male-to-male transmission, but male-to
female and female-to-male transmission of the
affected X can occur. X-linked traits are expressed
FOUR CHROMOSOMAL ABNORMALITIES

1. Turner Syndrome

 Turner syndrome is a chromosomal condition that alters development in females. Women with this condition tend to
be shorter than average and are usually unable to conceive a child (infertile) because of an absence of ovarian function.
 Other features of this condition that can vary among women who have Turner syndrome include: extra skin on the
neck (webbed neck), puffiness or swelling (lymphedema) of the hands and feet, skeletal abnormalities, heart defects
and kidney problems.

2. Klinefelter Syndrome

 also known as the XXY condition, is a term used to

describe males who have an extra X chromosome in
most of their cells.
 Instead of having the usual XY chromosome pattern
that most males have, these men have an XXY
pattern
 Even though all men with Klinefelter syndrome
have the extra X chromosome, not every XXY male
has all of those symptoms.

3. Down Syndrome/ Trisomy 21

 Decreased muscle tone at birth

 Excess skin at the nape of the neck
 Flattened nose
 Upward slanting eyes
 Small ears
 Small mouth
 Wide, short hands with short fingers
 Separated joints between the bones of the skull
 Single crease in the palm of the hand
 White spots on the colored part of the eye
  A genetic disorder and the most common autosomal chromosome abnormality in humans, where extra genetic
material from chromosome 21 is transferred to a newly formed embryo.
 These extra genes and DNA cause changes in development of the embryo and fetus resulting in physical and mental
abnormalities.
 Each patient is unique and there can be great variability in the severity of symptoms.
 In patients with Down syndrome, an error occurs in the coming together of chromosome 21.
 .The extra genetic material is responsible for the developmental abnormalities that occur.
 Instead of 46 chromosomes plus two sex chromosomes, there are 47.

4. Cri du chat or Cry of the Cat syndrome

a) Karyotype (G banding) b) Individual with Cri-du-chat syndrome

 Cri du chat syndrome (CdCS or 5p-) is a rare genetic disorder in which a variable portion of the short arm of
chromosome 5 is missing or deleted (monosomic).
 Symptoms vary greatly from case to case depending upon the exact size and location of the deleted genetic material.

Common symptoms include a distinctive cry that resembles the mewing of a cat, characteristic facial features, slow growth,
and microcephaly, a condition that indicates that head circumference is smaller than would be expected for an infant's age and
sex.

SESSION 14:
CARBOHYDRATES

Definition:
 The term carbohydrate originally referred to hydrates of carbon because the general formula of these compounds was
CnH2nOn or Cn(H20)n
 However, some materials with this general formula are not carbohydrates, and some carbohydrates don't have this
general formula
 These are compounds which are either polyhydroxyaldehydes or polyhydroxyketone or a compound which yields
either or both of these upon hydrolysis
 Are a product of photosynthesis, where inorganic carbon dioxide becomes organic carbon with the utilization of solar
energy, accompanied by the release of oxygen gas
 Saccharides - building blocks / fundamental sub-units of carbohydrates

Storage forms of Carbohydrate:

Glycogen - storage form of carbohydrate in animal tissue which is found in liver and muscle
Starch - storage form of carbohydrate in plant tissue

Biological Significance:
1.) Carbohydrates are the main source of energy in the form of ATP.
2.) Other carbohydrate metabolic products are used in the synthesis of other types of compounds.
Metabolism - process of breaking down into simplest forms
e.g. synthesis of fatty acids and amino acids
3.) Help in the breakdown of food stuff by acting as catalyst or promoters of oxidation.
CLASSIFICATION OF CARBOHYDRATES:
a. Simple sugars / monosaccharide
b. Oligosaccharides
c. Polysaccharides

A. SIMPLE SUGARS/MONOSACCHARIDES - these are CHO's that cannot be hydrolyzed to simplest forms. It has single
polyhydroxy aldehyde or ketone unit with general formula CnH2nOn

I. According to functional group present

1. Aldose - presence of an aldehyde group
2. Ketose - presence of a ketone group

II. Based on the no. of C-atoms in the molecule

a. Triose - (C3H6O3) - not found free in nature but as products of carbohydrates metabolism.
 simplest monosaccharide
o glyceraldehydes or glycerose (Aldose)
o dihydroxyacetone (ketose)

b. Tetroses - 4-C monosaccharide

- (C.H.O.)
o Erythrose - (aldose)
o Threose - (aldose)
o Erythrulose - (ketose)

c. Pentoses - 5-C monosaccharide

o Ribose: constituent of RNA
o Arabinose: present in cherries and seen in glycoproteins of the body.
o Xylose: is seen in proteoglycans
o Lyxose: occurs only rarely in nature, as a component of bacterial glycolipids
o Xylulose: is an intermediate in uronic acid pathway

d. Hexoses - 6-C monosaccharide

- are the most important monosaccharides found in plants.
- they are the first detectable sugars in plant
- glucose and fructose - occur in free state in plants, sweet fruits, honey and inverted sugar (came from the
hydrolysis of sucrose yielding equimolar quantities of glucose and fructose)
o Allose
o Altrose
o Glucose: most abundant monosaccharide in nature
 *3 epimers of glucose (Epimers: differ from each other only at a single carbon atom)
 Mannose: 2nd carbon
 Galactose: 4th carbon
 Idose: 5th carbon
o Mannose
o Gulose
o Idose
o Galactose
o Talose
o Fructose (ketohexose)

o Heptoses - 7-C sugar form a vital importance in the glucose metabolism of animals and in the photosynthesis
process of plants.
f. Octose - 8 - carbon sugar isolated from avocado pulp
- D-glycerol -D-manno octulose

B. OLIGOSACCHARIDES - carbohydrates which on hydrolysis will yield 2-10 units

 Disaccharides (C12H22O11) (considered as the most abundant oligosaccharide)
1.) Reducing disaccharides - those which contain a free aldehyde or ketone group

a. Lactose

o milk sugar least soluble and least sweet of all sugars

o B-D galactose + B-D glucose
o cow's milk - 4-5% lactose
o human-7-8% lactose

b. Maltose

o malt sugar found in germinating grains and in malt; are among the series of substances formed by the hydrolysis of
starch
o B-D glucose + B-D glucose

c. Cellobiose

o obtained from the hydrolysis of cellulose from wood & cotton

o B-D glucopyranose + B-D glucopyranose

Test for Reducing Sugar:

(+) result

1. Barfoed's test brick red ppt.

2. Benedict's test brick red ppt.

* BENEDICT'S REAGENT - CONSTITUENTS "3C's"

 C-Copper Sulphate (provides cuprous ions)

 C- Citrate (Sodium Citrate) (stabilizes reagent)
 C-Carbonate (Na Carbonate) (provides alkaline medium for benedicts reaction)

3. Nylander's test black ppt.

4. Fehling's test brick red ppt.

Fehling's A Fehling's B

- CuSO4 - NaOH
- Rochelle's salt

(+) results - formation of brick red ppt.

CuSO4

Principle: Reduction is due to the reduction of Cupric SO4 - Cuprous oxide

 Fehling's Test - basis of clinical determination of glucose in blood or urine

Detection of Reducing Sugar in:

1. Fischer Formula - free aldehyde or ketone group in the structure

2. Haworth Structure - presence of an OH group at C1 (aldose) or C2 (ketose)

2.) Non-reducing sugars:

o are those whose aldehyde groups are involved in the linkage

o no free aldehyde or ketone group

a. Disaccharide/s

 Sucrose
o a.k.a table sugar, cane sugar, saccharose, beet sugar
 o most widely distributed in nature
o obtained commercially from sugar cane and sugar beets
o a-D glucose + B-D fructose
o connected by alpha 1-2 glycosidic linkage.

* Honey contains invert sugar. Invert sugar is sweeter than sucrose.

b. Trisaccharide/s-C18H32016

 Raffinose
o forms in sugar beets
o made up of glucose, fructose and galactose

c. Tetrasaccharide/s - yield for monosaccharide molecules on hydrolysis

 Stachyose-fructose + glucose + 2 molecules of galactose

C.) POLYSACCHARIDES - carbohydrates which on hydrolysis will yield several monosaccharides units.

- more than 10 saccharides

- more complex, high ocular weight

1- CELLULOSE GROUP:

 Cellulose
o most abundant organic compound making up about 50% or more of the carbon vegetation
o the purest source is cotton
o serves as roughage for the evacuation of the bowel
o has D-glucose residues in 1-4 linkage and is not hydrolyzed neither a orb amylase and not digested
by vertebrate except by ruminant animals, in which cellulase secreted by bacteria, fragments
cellulose into D-glucose
 Pentosans - (C5H8O3)n - x H2O - long chain of pentose units

1. Xylans - found in wood, straw, rice bran and corn cobs

2. Arabans - found in gum Arabic, mucilage and fruit juices.

 Hexosans - (C6H10O3)n- x H2O

1. Glucosan - starch yielding glucose

2. Agar agar

 Dextran
 Polysaccharide produced by certain microorganisms when grown on sugar media
 Made up of units of D-glucose molecule having glycosidic linkages
 Synthesized by Leuconostoc dextranicum
 Used as blood extender due to its high viscosity, osmotic pressure, low disintegration and utilization
 Hexopentosans

1. Pectins

 Colloidal carbohydrates
 Responsible for jellying properties of fruits
 Commercially prepared from apples and lemons
 On hydrolysis it yields arabinose, galactose, acetic acid, methyl alcohol and galacturonic acid
SESSION 15:
 HETEROPOLYSACCHARIDES
 Polysaccharides which on hydrolysis yield mixtures of heteropolysaccharides and derived products
 2 MAIN GROUPS:

a. Neutral mucopolysaccharides

o Made up of N-acetyl-hexosamine and hexose

o Example those occurring in bacteria and 'so-called "mucoids" including important immunological, specific blood group
substances

b. Acid mucopolysaccharides

o Contain hexuronic acid, sulfate or phosphate

 HYALURONIC ACID
 Made up of d-glucoronic acid, d-glucosamine and acetic acid in equimolecular amounts
 Serves as a cementing substance in the tissues which allows the passage only of the metabolites but not the infecting
microorganisms. However it is fragmented by hyaluronidase ("'spreading factor") from bacteria, allowing the diffusion
and spread of bacterial infection

 HEPARINE (heparin)
 An important acid mucopolysaccharide secreted by the lung tissues and certain types of cells lining the arterial blood
vessels
 Powerful inhibitor of blood clotting, thus preventing intravascular coagulation

*Five Glycosaminoglycans (GAG) /Mucopolysaccharides

 Hyaluronic acid
o Contains D-glucoronic acid + glucosamine
o It is sulfate free GAG
o Present in high concentration in testes, seminal fluid, & in certain snake and insect venoms.
o Serves as a lubricant and shock absorbent in joints.
o Hyaluronidase enzyme of semen degrades the gel around ovum & allows effective penetration of sperm into ovum

 Chondroitin sulfate
o Most abundant GAG in the body.
o Contains D-Glucoronic acid + Galactosamine
o It is present in cartilages, tendons, ligaments, and aorta which helps to maintain their shapes.
o Osteoarthritis( the amount of Chondroitin sulfate decreases while that of Hyaluronic acid increases)

 Keratan sulfate
o Contains D-galactose + Glucosamine
o Only GAG with no uronic acid
o Found in cornea and tendon
o 2 types
 Keratan sulfate 1 - found in cornea
 Keratan sulfate II - found in skeletal muscle
o Maintains the corneal transparency

 Dermatan sulfate
o Contains L-iduronic acid + Glucosamine
o Found in Skin, Blood vessels ,Heart valves & Cornea
o It’s presence in the Sclera maintains overall shape of the eye.
o Major GAG synthesized by arterial smooth muscle cells
 Heparin
o Contains D-glucoronic acid + glucosamine
o It is the only intracellular GAG
o It is an anticoagulant

Naming of Polysaccharides:

Examples

a. hexose - it is termed hexosan

b. starch - yielding glucose is known as glucosan

c. starch - yielding fructose is known as fructosan

Differences between sugar and starch

Properties Sugar Starch

Physical State crystalline solid amorphous solid
Taste Sweet tasteless
Solubility in water soluble slightly soluble in H2 to insoluble in cold
H soluble in hot H2O - gel

PROPERTIES OF CARBOHYDRATES:

I - PHYSICAL

A. Monosaccharides and Oligosaccharides:

1. White crystalline solids

2. Soluble in water

3. Optically active - due to the presence of a chiral carbon

a. Dextrorotatory - right (+)

b. Levorotatory - left (-)

4. Sweet taste

B. Polysaccharides are:

1. Amorphous solid

2. Low molecular weight form a colloidal dispersion in H2O

3. High molecular weight are insoluble in H2O (eg. Cellulose)

4. Not sweet / no flavor at all

II - CHEMICAL

A. Hydrolysis

1. Disaccharides: C6H12O11+ H2OC6H12O6 + C6H12O6

Lactose + H2O glu + gal

Sucrose + H2O glu + fru

Maltose + H2O glu + glu

Cellullose + H2O 2 ß-glu

 2. Starch Hydrolysis

STARCH –AMYLODEXTRIN – ERYTHRODEXTRIN – ACHRODEXTRIN – MALTOSE - GLUCOSE

Agents of Hydrolysis:
1. Heat
2. Acid
3. Bases
4. Enzymes

B. Reducing Property or Action of Carbohydrates:

 Attributed to a free aldehyde or ketone group
 Reducing Tests
 All are oxidized into acids CHO-COOH
 Reducing sugars are called as such because
Eg. Fehling's Test - Reduction is due to the reduction of Cupric SO4-Cuprous oxide

C. Fermentation
o Complex process which involves the breaking down of complex substances with the aid of biological catalysts called
enzymes
o Ethyl alcohol + Co, are the products of fermentation

Eg. Fermentation of Sugar:

i. C12H22O11 + H2O Invertase C6H12O6 + C6H12O6

ii. C6H12O6 Zymase 2C5H5OH + 2CO2

D. Osazone Formation

Reagent: Phenylhydrazine
(+) Result: yellow crystals or yellow ppt.
Osazone formation aids in the identification by:

a. Each osazone has a definite crystal formation

 Maltose - petal-shaped
 Lactose - powder puff-shaped
 Galactose - rhombic-plate
 Glucose, Fructose, and Mannose - Broom shaped

b. Each osazone has a definite melting point

c. Length of time for osazone formation

3 Reactions Involved in Osazone Formation:

a. Formation of glucosazone
b. Formation of fructosazone
c. Formation of mannosazone
 Mannose - first one to form an osazone

E. Molisch Test

o a general test for carbohydrates except trioses and tetroses which lack the minimum requisite of 5C atoms
o Reagent: a-naphthol + conc. H2SO4
o (+) Result: violet ring at the junction

F. Oxidation

o Aldehyde - Acid

G. Reduction

o Aldehyde - Alcohol