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1) Glycolysis is an almost universal pathway for extraction of the energy available from
carbohydrates, shared among prokaryotes and eukaryotes, aerobes and anaerobes
alike. By your understanding, discuss it and classify the 10 enzymes into six categories
as you learnt.
Glycolysis is an almost universal pathway for extraction of the energy available
from carbohydrates, shared among prokaryotes and eukaryotes, aerobes and anaerobes
alike. In anaerobes, glycolysis is the only significant source of energy from
carbohydrates. In aerobic organisms, considerably more energy can be harvested
downstream from glycolysis in the citric acid cycle. Glycolysis produces energy in the
form of ATP and NADH.
The intermediate products of glycolysis can be used for other pathways.
-For instance, during active energy production, pyruvate, the product of
glycolysis, enters the citric acid cycle. In the absence of oxygen, fermentation
converts pyruvate into lactic acid.
-When energy levels are high, glucose-6-phosphate, an intermediate of
glycolysis, can enter glycogenesis to be stored as glycogen, or enter PPP to
provide R5P for nucleic acid synthesis. PPP also produces GAP, an intermediate
for glycolysis.
- Glyceraldehyde-3-phosphate, which is produced by photosynthesis, is
also a glycolytic intermediate, so it can be directed from this anabolic pathway
into glycolysis when energy is needed
-Glycogenolysis can degrade glycogen to Glucose-6-phosphate for
Glycolysis to go on in situation where there is shortage of glucose.
(pathway glycolysis xem cu 7)
Step 1: Hexokinase: transferase
Step 2: phosphoglucoisomerase: Isomerase
Step 3: phosphofructokinase: transferase
Step 4: aldolase: lyase
Step 5: tri-isomerase: isomerase
Step 6: triose phosphate dehydrogenase: oxidoreductase
Step 7: phosphoglycerokinase: transferase
Step 8: phosphoglyceromutase: isomerase
Step 9: enolase: lyase
Step 10: pyruvate kinase: transferase
2) You have learnt methods to determine soluble protein, total soluble sugar, calcium
content and enzymatic activity (Bromelain) in a sample. Choose one and describe the
principle, chemicals, experimental design and analysis for the final result.
Hartree Lowry method to determine soluble protein in a sample.
1. Principle: The protein content in a sample is determine based on the quantitative reaction
between copper ion and phosphomolybdic/phosphotungtic acid (Folin Cioucalteu reagent)
with protein.
Glucose
oxidized
glycolysis,
ku
draw
thi
nhng vit ra
cho c hiu hnh cho d thuc d vis
by
an
energy-
(~90%) and kidneys from pyruvate, lactate, glycerol, amino acids, and
TCA cycle intermediates.
Gluconeogenesis pathway consists of 11 steps which are reverse of
glycolysis and there are 3 bypass steps- which use different enzymes and
mechanisms compare with glycolysis. (ci bng l so snh bypass steps)
(hnh cho d hnh dung Gluconeogenesis)
Storage of glycogen:
In muscle forms as cytosolic granules with a diameter of 10 to 40 nm - the same
as the size of ribosome.
Concentration (liver cells) > concentration (muscle cells)
Total amount of glycogen in muscle cells > in the liver.
In the uterus, glycogen is stored during pregnancy to nourish the embryo.
In vagina, glycogen is secreted and then converted into lactic acid to maintain the
acidic environment in order to protect vagina from outside bacteria infections.
In kidneys, brain and white blood cells.
Glycogenesis
Excess of glucose
Glycogen is synthesized from glucose-6-phosphate.
Glycogen can either enter into pentose-phosphate pathway or contribute to the
process of glycolysis.
Involves 3 steps:
1. Synthesis of glucose-1-phosphate
2. Synthesis of UDP-glucose
3. Synthesis glycogen from UDP-glucose
Glycogenesis is the formation of glycogen from glucose. Glycogen is synthesized
depending on the demand for glucose and ATP (energy). If both are present in relatively
high amounts, then the excess of insulin promotes the glucose conversion into glycogen
for storage in liver and muscle cells. (It means that insulin inhibits glycogenolysis by
inhibiting Glycogen phosphorylase enzyme activity.)
In muscle cells, glycogen degradation serves to provide an immediate source of glucose6-phosphate for glycolysis, to provide energy for muscle contraction.
In liver cells, the main purpose of the breakdown of glycogen is for the release of
glucose into the bloodstream for uptake by other cells. The phosphate group of glucose-6phosphate is removed by the enzyme glucose-6-phosphatase, which is not present in
myocytes, and the free glucose exits the cell via GLUT2 facilitated diffusion channels in
the hepatocyte cell membrane.
9) Fatty acids are an important energy source as their yield over twice as energy as an
equal mass of carbohydrate or protein. The two main pathways of fatty acid
metabolism are oxidation and fatty acid synthesis. oxidation result in the formation
of reduced cofactors and acetyl-CoA molecules, which can be further catabolized to
release free energy. By what you have learnt, discuss a round of oxidation and focus
on the changing of each reaction, the energy yield, and name of enzymes.
The activated fatty acid is called a fatty acyl-coenzyme A, or fatty acyl-CoA.
-In the first step of oxidation, an acyl-CoA dehydrogenase catalyzes the
oxidation of the acyl group, resulting the formation of a double bond between carbons
two and three. The two electrons removed from the acyl group are transferred to an FAD
prosthetic group. These electrons are transferred to ubiquinone through a series of
electron transfer reactions.
-In the second step of oxidation, a hydratase adds a molecule of water across
the double bond produced in the first step.
-In the third step of oxidation, another dehydrogenase catalyzes the oxidation
of the hydroxyacyl group. In this case, NAD+ is the cofactor. The fourth and final step
of oxidation is called thiolysis. In this step, a thiolase catalyzes the release of acetylCoA from the ketoacyl-CoA.
Energy yield:
One round of oxidation yields three productsone ubiquinol cofactor, one NADH
cofactor, and one molecule of acetyl-CoA.
-During oxidative phosphorylation, 1 ubiqunol -> 2 ATP; 1 NADH -> 3ATP, 1
actyl-CoA -> 12ATP => total 17 ATP, and the net ATP is 15 (2 was used for fatty acid
activation)
10)
Call the names of below fatty acids and calculate the energy yield followed by
equation of oxidation
that is insufficient to meet the growth needs of the body and the majority that is synthesized is
cleaved to form urea. Methionine is required in large amounts to produce cysteine if the latter
amino acid is not adequately supplied in the diet. Similarly, phenylalanine is needed in large
amounts to form tyrosine if the latter is not adequately supplied in the diet.
12) Why do doers have to pay attention to the safety concern in the Biochemistry Lab? List
out the hazardous factors that doers may encounter. List out all the concentration units
that are commonly used in the Biochemistry Lab.
Safety concern is very important in the Biochemistry Lab. When perform tasks in the lab,
doers have to due with many toxic, erosive strong chemicals, as well as hard to handle
machines, and electricity. The safety concerns are to help the doers yield the best result
for their experiments in the safe way.
-Toxic chemicals, strong acids, base.
-Explosive, exothermal reactions.
-Burning of chemicals
-Fire
-Electric shock
-Production of hazardous gas.
-Breakage of instruments.
-Malfunctioning instruments.
Concentration units that are commonly used in the Biochemistry Lab:
n(mol solute)
-Molarity (M) M =
V (L solution)
n(mol solute)
kg( solvent )
n mol equivalents
N=
L solution
-Molality m=
-Normality
- g/mL
mi ngi gp , chnh sa b sung cu ny dm vi :3
13) Show your acknowledgement and evaluation upon the presence and roles of
Cholesterol ( in total, in HDL and in LDL forms) in human body
HDL cholesterol:
HDL stands for high-density lipoproteins, which is known as the "good" cholesterol.
HDL cholesterol carries the cholesterol away from arteries in into liver, thereby preventing heart
disease.
A high HDL level helps to preserve brain cell function and prevent against mental decline in the
elderly.
High levels of HDL are also used as a tool for diagnosing metabolic syndrome, thereby preventing
heart attack.
HDL contains antioxidant molecules that may prevent LDL from being changed into a lipoprotein,
which helps prevent heart disease.
LDL cholesterol:
LDL stands for low-density lypoproteins which is also known as the "bad" cholesterol for human
health;
High levels of LDL in blood stream, it deposits the cholesterol into arteries, which can cause
blockage and lead to heart attack.
LDL cholesterol an add to the build-up of plaque in arteries and increase risk of getting coronary
heart disease.
In total:
Total cholesterol is the sum of HDL cholesterol, LDL cholesterol and 20% of the triglyceride value.
o Total cholesterol = HDL cholesterol + LDL cholesterol + (Triglyceride/5)
- In case of setting ( or low) amount of total cholesterol in body, we have to consider carefully that:
o If high total cholesterol level is caused by high HDL level, we do not need to worry. Because HDL
cholesterol is good for our health. It transports cholesterol form the tissue back to the liver where
it is secretes in the bile.
o If a high total cholesterol level is caused by high LDL level, we need to pay more attention to our
diet, our inputs in order to avoid heart disease, obesity
In brief, both HDL and LDL play an important role in human body. Total cholesterol has the relation with HDL and
LDL. The higher or lower of HDL and LDL affects the level of total cholesterol, which has effects on human body.
-
14) What are the most commonly known functionalities of fatty acids in human body?
First, fatty acids are building blocks of phospholipids and glycolipidsamphipathic molecules that are important components of biological membranes.
Second, many proteins are modified by the covalent attachment of fatty acids,
which targets them to membrane locations.
Third, fatty acids are fuel molecules. They yield over twice as much energy as an
equal mass of carbohydrate or protein. They are stored as triacylglycerols (also called
neutral fats or triglycerides), which are uncharged esters of fatty acids with glycerol.
Fatty acids mobilized from triacylglycerols are oxidized to meet the energy needs of a
cell or organism.
Fourth, fatty acid derivatives serve as hormones and intracellular messengers.
Cholesterol is a lipid with a hydrocarbon tail linked to the steroid (built from four linked
hydrocarbon rings) at one end, and a hydroxyl group attached at the other end. It is a
common component of cell membrane and a precursor for the synthesis of other steroid
such as vertebrate sex hormones (estradiol and testosterone). Intracellular messengers are
generated by activated receptors on cell membrane to transmit information into the cell,
diacylglycerol and triphosphoinositol are examples of intracellular messengers.
15) What is the closed correlation of cholesterol and fatty acids?
Cholesterol and fatty acids are 2 forms of lipid, which circulate in your bloodstream.
- They are both necessary for life itself. Cholesterol is necessary for building, maintaining key parts of our cells and
making several essential hormones (estrogen, progesterone, testosterone). While triglycerides, which are chains of
high-energy fatty acids, provide energy for tissues to function.
- High levels of both cholesterol and fatty acids can cause heart disease.
- Cholesterol includes HDL, LDL and total cholesterol. Total cholesterol is the sum of HDL, LDL and 20%
triglycerides of value. It is clarified by formula:
Total cholesterol= HDL cholesterol + LDL cholesterol+ Triglycerides/5
- Triglycerides has 2 sub-classes, which relate to cholesterol
18) Discuss about roles of the amino acid carbon skeletons in the deamination process.
The carbon skeletons of the -ketoacids are converted to common intermediates of energy
producing, metabolic pathways. These compounds can be metabolized to CO2 and water,
glucose, fatty acids, or ketone bodies by the central pathways of metabolism.
The carbon skeletons of the -ketoacids, from deamination, results in the formation of seven
products: pyruvate, acetyl-CoA, acetoacetyl-CoA, -ketoglutarate, suc-CoA,
fumarate andoxaloacetate (Kreb Cycle).
Amino acids are divided into glucogenic and ketogenic amino acids:
Ketogenic amino acids (leucine and lysine) lead to the formation of acetyl-CoA and
acetoacetyl-CoA.
Amino acids called keto- and glucogenic amino acids (isoleucine, phenylalanine,
tyrosine and tryptophan) have two degradation products one of them being
glucogenic and second one ketogenic.
19) ATP synthesis via oxidative phosphorylation occurs via two separate systems in the
mitochondrion. What are they? Briefly summarize the involving steps in the ATP
synthesis.
Oxidative phosphorylation is the synthesis of ATP from ADP (phosphorylation), that
occurs when NADH and FADH2 are oxidized throughout electron transport chain (oxidation).
Inner membrane of mitochondria is the site of oxidative phosphorylation in eukaryotes
A. Oxidation step: electron transport chain
NADH + H+ + O2 NAD+ + H2O
FADH2 + O2 FAD + H2O
B. Phosphorylation step
ADP + Pi ATP
The electron transport chain is series of protein complexes embedded in mitochondrial
membrane.
This chain is consisting of 4 complexes and ATP Synthesis.
4 complexes are:
- NADH (Complex I)
- FADH2 (Complex II)
- Cytochrome b-c (Complex III)
- Cytochrome oxidase (Complex IV)
ATP synthesis via oxidative phosphorylation occurs via two separate systems in the
mitochondrion.
1. Electrons are transported via numerous membrane-bound carriers from NADH to O2.
During these reactions, a proton gradient is formed across the mitochondrial inner membrane.
2. The proton-motive force in the gradient is then harnessed to produce ATP.
- Mitochondrial structure plays a critical role in forming and utilizing the proton gradient
to synthesize ATP.
- Protons are pumped from the matrix across the inner membrane into the intermembrane
space.
- ATP is synthesized in the matrix, as protons flow back through the membrane.
Briefly summarize the involving steps in the ATP synthesis (1)
A complex channel protein known as ATP synthase, also called complex V, is the final enzyme in
the oxidative phosphorylation pathway. It allows the controlled flow of protons back into the
matrix while, at the same time, harnessing the free energy of proton flow to convert ADP and
phosphate into ATP. ATP can be thought of as a form of portable free energy, or an energy
currency, that can be used for energy-requiring reactions elsewhere in the cell. This is analogous
to water flowing through the turbines of a dam to generate electricity.
NADH: The flow of 2 e (to reduce1 oxygen atom O) through complexes I, III & IV
results in the translocation of 3 ATPs
FADH2: The flow of 2 e through complexes II, III & IV results in the translocation of 2
ATPs
Briefly summarize the involving steps in the ATP synthesis (2)
Synthesis of ATP is coupled with the oxidation of NADH and the reduction of O2. There are
three key steps in this process:
1. Electrons are transferred from NADH, through a series of electron carriers, to O2. The
electron carriers are proteins embedded in the inner mitochondrial membrane.
2. Transfer of electrons by these carriers generates a proton (H+) gradient across the inner
mitochondrial membrane.
3. When H+ spontaneously diffuses back across the inner mitochondrial membrane, ATP is
synthesized. The large positive free energy of ATP synthesis is overcome by the even larger
negative free energy associated with proton flow down the concentration gradient.
20) Briefly discuss about relation of the Pentose phosphate pathway (PPP) and Glycogen
metabolism.
Both glycogen metabolism and pentose phosphate pathway is considered as glucose anabolism
processes. The relation of PPP and glycogen metabolism is briefly discuss by the following
figure.