Bangalore University

VI Semester, B.Sc., Examination, June 2019

CHEMISTRY Paper - VIII (BIOCHEMISTRY)

Scheme of Evaluation

Amk manjunath krishnappa

Dept.of chemistry
GFGC chickballapura

PART - A

Answer any eight of the following questions. Each question carries two
marks (8X2=16)

1. Mention the contributions of the following scientists to the

development of Biochemistry: a) Hans Krebs b) Fredric Sanger

a) Hans Krebs:
Elucidated the citric acid cycle and urea cycle (1M)

b) Fredric Sanger: (1M)

1) Proposed structure of insulin
2) Base sequence of nucleic acids [any one point]

2. Write the Zwitter ionic structural form of glycine. (2M)

3. What is Michaelis–Menton constant? Mention its significance. (2M)

Michaelis-Menton constant (Km) is defined as the concentration of the substrate at

which the velocity of the reaction is half the maximum velocity. The Km value signifies
the affinity of an enzyme for the substrate. The value is characteristic of an enzyme.

Significance of Km: Km value signifies the affinity of an enzyme for the substrate.
Smaller Km value indicates that there is high affinity of the enzyme for the substrate.
This shows that even at low [S], the enzyme is half saturated or ½ Vmax is reached. A
high Km value indicates the less affinity of enzyme for the substrate. This mean to attain
½ Vmax, high substrate concentration is required.
 4. Write Haworth structure of β-D -Galactosamine. (2M)

5. What is energy coupling? Give an example.

ENERGY COUPLING: (1M)

The phenomenon where a thermodynamically unfavourable [endergonic reaction with

∆G= +ve] reaction can be driven in the forward direction by coupling it to a highly
favourable [exergonic reaction with ∆G= -ve] reaction, is called energy coupling.

Eg: (1M)

a. Phosphorylation of glucose is an endergonic / unfavourable reaction

Glucose + Pi Glucose-6-phosphate +H2O; ∆G= +13.8 KJ/mol
b. ATP hydrolysis is a highly exergonic / favourable reaction
ATP + H2O ADP + Pi ; ∆G= -30.5 KJ/mol
c. When reaction a is coupled with reaction b , the overall reaction is exergonic
/favourable reaction.
Glucose + ATP Glucose-6-phosphate +ADP ; ∆G= +13.8 - 30.5 = -16.7
KJ/mol

6. Mention any two termination codons. (2M)

TERMINATION CODONS: [any two of the following]

Amber [UAG]
Ochre [UAA]
Opal [UGA]

7. Define saponification number of lipids. What is its significance?

(1 +1 M)
Saponification number is defined as number of miligrams of KOH required to saponify
one gram of oil or fat. i.e., to neutralise the fatty acids resulting from complete hydrolysis
of one gram of oil or fat.

Significance:
saponification value signifies the size of fatty acid in oil or fat. Smaller the saponification
value, higher is the molecular size of fatty acid.

8. Write the reaction of Urea cycle catalysed by arginase. (2M)

Arginase enzyme catalyses the final step of urea cycle in the cytosol of liver cells.
Arginase enzyme is a hydrolyase which catalyses the cleavage of arginine into urea and
ornithine.
9. Explain competitive inhibition with example. (1+1 M)

It is a type of reversible inhibition. In this type, the inhibitor resembles the natural
substrate in its structure. Hence, it competes with the substrate for the active site and
reversibly binds for the enzyme at the active site forming enzyme inhibitor complex (ES).
However the increase in substrate concentration decreases the enzyme inhibition.

For example:

For the enzyme succinate dehydrogenase (SDH), malonate is the inhibitor. Malonate
structurally resembles succinate (the actual substrate) and hence reversibly binds to the
active site of substrate. Similarly, thiourea is an inhibitor for urease as it resembles urea
in its structure.

10. What are nucleotides? Give an example. (1+1 M)

The condensation products of nucleoside with phosphoric acid are called nucleotides.
OR
The condensation products of a nitrogenous hetero cyclic base with a five membered
ribose sugar and phosphoric acid are called nucleotides.
Eg. AMP, ADP, ATP etc.

11. Write the structure of Alanyl serine. (2M)

12. Name four factors that stabilize tertiary structure of proteins. (2M)

The factors that stabilize the tertiary structure are

1. Hydrophobic interactions 2. Disulphide bond
3. Hydrogen bonds 4. Ionic interactions

PART - B
 Answer any nine of the following questions. Each question carries six marks
(9X6=54)

13. a. Write the principle and applications of Thin layer chromatography.

PRINCIPLE: (2M)
Differential affinity of the various components of the analyte towards the stationary
and mobile phase results in the separation of components.

APPLICATION: (2M)
 To check the purity of given samples.
 Identification of compounds like acids, alcohols, proteins, alkaloids, amines etc.
 To evaluate the reaction process by assessment of intermediates, reaction course etc.
 To keep a check on the performance of other separation processes.
(Any two 2)

b. Write any two biological role of water. (2M)

Water has dielectric constant (80): It reduces the attractive force between ions. It
can therefore dissolve a large variety of ionic and polar substances, making it the
universal solvent. In cells, many kinds and relatively large amounts of polar molecules
can dissolve or ionize in water because of its remarkable solvent qualities.

High heat of vaporization: The high heat of vaporization as a result of hydrogen

bonding of water enables humans and other animals to maintain a steady body
temperature even during intense physical activity or high metabolic rates or adverse
environmental conditions and thus water acts as heat buffer.

14. a. Write the partial structure of glycogen, give its biological importance.
How does it differ from Starch?

 Partial structure of Glycogen (2M)

α (1-6) linkage

α (1-4) linkage

 Biological importance: (1M)

Glycogen is a stored form of energy in liver and muscles which provide readily
available source of energy if blood glucose levels decrease.
  Difference between starch and glycogen (1M)

Starch is stored in plants as food reserve composed of two forms amylose and
amylopectin whereas glycogen is found in animals which is stored in liver and
muscle, it does not consists of amylose and amylopectin. It is structurally
similar to amylopectin but highly branched.

b. Give any two biological role of chitin. (2M)

 Chitin is the major component in the exoskeleton of insects.

 Chitin is the cell wall component of fungi.
 Chitin is present in combination with calcium carbonate to form hard structures
in invertebrates and fishes.
[Any Two]

15. a. What are fatty acids? Explain the classification with examples. [4M]

The long chain monocarboxylic acids which contain even number of carbon atoms are
called fatty acids.

Classification:
 Based on nature of chemical bonds, fatty acids are classified into saturated and
unsaturated fatty acids.
i. Saturated fatty acid: They contain single bonded carbon chain. The general
formula: CH3(CH2)nCOOH. Eg: Butyric acid, Caproic, Palmitic acid, Stearic
acid etc.
ii. Unsaturated fatty acids: They contain one or more double bonds.
eg , Oleic acid, linoleic, linolenic and arachidonic acid

 Fatty acid can also be classified as

i. Essential Fatty acids. Polyunsaturated fatty acids which are essential for
normal health but not synthesised in humans and hence they must be in the
diet are called essential fatty acids. eg, linoleic, linolenic and arachidonic acids
ii. Non-essential fatty acids: Essential for normal health but are synthesised
in humans hence need not be present in the diet. Eg. Steric acid, palmitic acid
or any other

b. Define liposomes. Give any two applications of it. [1+1 M]

Liposomes are hollow spheres or vesicles with a lipid bilayer membrane, enclosing an
aqueous cavity. The hydrophilic regions of outer and the inner membrane face the
external and internal aqueous regions respectively while the hydrophobic regions are
directed towards one other.

APPLICATION of Liposomes:
 Liposomes are used for administration of nutrients.
 Liposomes are used as vehicles for drug and vaccine targeting.

16. a. What are hormones? Give the biological functions of

 (i) Glucagon (ii) Oxytocin (iii) Progesterone. (4M)

Hormones are chemical messengers that are secreted directly into the blood, which
carries them to organs and tissues of the body to exert their functions. There are
many types of hormones that act on different aspects of bodily functions and
processes. Some of these include:

 Development and growth

 Metabolism of food
 Sexual function and reproductive growth and health
 Cognitive function and mood
 Maintenance of body temperature and thirst

(i) Glucagon- It increases blood glucose level by glucogenesis/ It stimulates the

release of lipid from adipose tissue (lipolysis)/ Stimulates formation of ketone bodies.

(ii) Oxytocin-Causes contraction of the smooth muscles of the uterus during child
birth & Causes contraction of the smooth muscles of the mammary glands resulting
in the secretion of milk.

(iii) Progesterone- It is essential for maintenance of pregnancy/ It promotes the

proliferation of the uterine mucosa for implantation of the fertilized ovum. (Any one)

b. Write a note on denaturation of proteins. (2M)

Denaturation of proteins is a phenomenon where proteins lose their tertiary or
quaternary structures present in their native state by the effect of heat, pressure or
chemicals.
Denaturation results in the loss of activity of proteins. Denaturation can be reversible
or irreversible.

17. a. Explain the classification of amino acids based on polarity of side chain
with examples. (4M)

Based on polarity, amino acids are classified into four types.

1. Amino acid with non-polar or hydrophobic side chain. eg: Alanine, phenylalanine
2. Amino acid with uncharged polar or hydrophilic side chain. eg: Serine, Cysteine,
etc.
3. Amino acid with negatively charged polar side chain. Eg:, Aspartic acid, Glutamic
acid
4. Amino acid with positively charged polar side chain. Eg: Lysine, Histidine, and
Arginine

b. Write a note on primary structure of proteins. (2M)

The linear covalent sequence of amino acids of a protein is called its primary
structure. Every primary structure has an N-Terminal end [amino] and a C-Terminal
end [Carboxy]. The primary structure represents different amino acids and their
peptide linkage in a protein. Primary structure decides the chemical and biological
properties of proteins and their 3-D conformations. The primary structure of proteins
dissolved in water cannot be disrupted by heating above 80ºC.

18. a. (i) Name any two aromatic amino acids.

 (ii)Name any two hydroxyl group containing amino acids.
(2M)

(i) aromatic amino acids

 Phenylalanine.
 Tryptophan.
 Tyrosine.
 Histidine

(ii) Hydroxyl group containing amino acids.

 serine and threonine [contain aliphatic hydroxyl group]
 Tyrosine [has a hydroxyl group in the aromatic ring, making it a phenol
derivative].

b. Define isoelectric pH. Write the ionic form of alanine at pH 4.1. (2M)

The pH of the solution at which the amino acid exist in in the form of zwitter ion and
does not move either to the cathode or to the anode is called isoelectric point or
isoelectric pH. Each amino acid has the characteristic isoelectric point.

c. How do amino acids reacts with alcohol? Give equation. (2M)

Amino acids react with alcohols in presence of dry HCl gas, to form esters. Dry HCl
not only acts as a catalyst but also reacts with the amino acid.

19. a. Give brief account of Koshland’s induced fit theory of enzyme

catalysis. (2M)

The formation of enzyme substrate [ES] complex is necessary for enzyme catalyzed
reaction to occur .There are two theories to explain the formation of ES complex. The
mechanism by which most of the enzymes form ES complex is Induced Fit theory
proposed by Koshland in 1958. According to this theory, The active site of the enzyme
is not rigid, but flexible. The binding of the substrate induces a conformational change
in the enzyme such that the active site of the enzyme assumes a shape that is
complementary to the substrate.

When the substrate molecules bind to the enzyme

molecule, a change is brought about in the active site to precisely fit the transition
state (induced fit). This induced fit hold the substrates at the correct angle for the
reaction to take place.

b. Effect of temperature on enzyme catalysed reaction. (2M)

All enzymes are functional at a range of temperature which has a minimum, optimum
and maximum value. Enzymes are most effective at an optimum temperature which
is approximately near the body temperature. At temperature above the maximum
temperature, they lose their activity. At temperatures below the minimum, they are
not activated and the enzyme catalysed reactions are slow or not initiated. The
enzyme activity is maximum at the optimum temperature but is its minimum at the
minimum and maximum temperatures of the range.

c. Active site of an Enzyme. (2M)

The active site of an enzyme is the region that binds to the substrates. It also
contains the residues that directly participate in the making and breaking of bonds. It
occupies a relatively small portion of the enzyme. The sequence of amino acids that
form the active site is responsible for the enzyme specificity.

20. a. Calculate the number of ATP molecules produced by the oxidation

of acetyl CoA in TCA cycle. (4M)

The number of ATP molecules produced by the oxidation of Acetyl CoA in TCA cycle
 Step in TCA Enzyme Equivalent No. of ATP’S
Reaction
cycle involved produced produced

Isocitrate to Isocitate
3 NADH+H+ 3
α- ketoglutarate dehydrogenase

α- ketoglutarate α- ketoglutarate
4 NADH+H+ 3
to Succinyl CoA dehydrogenase

Succinyl CoA to Succinate

5 GTP 1
Succinate. thiokinase

Succinate to Succinate
6 FADH2 2
Fumarate. dehydrogenase

Malate to Malate
8 NADH+H+ 3
Oxaloacetate. dehydrogenase

Total ATP produced from one acetyl CoA/ one round of TCA cycle 12

b. Write equation for the conversion of fumarate to malate. (2M)

Equation for conversion of Fumarate to Malate.

Fumaric acid adds a water molecule along the carbon-carbon double bond to form
malic acid in the presence of enzyme Fumarase.

21. a. Describe the role of carnitine in the metabolism of fatty acid.(2M)

Carnitine is a mitochondrial membrane carrier which is responsible for the

transport of activated fatty acids [fatty acyl CoA] across the mitochondrial membrane
for β-oxidative breakdown.

Transport of Acyl CoA to mitochondria

The inner mitochondrial membrane is impermeable to fatty acids. Thus, a specialized
shuttle namely Carnitine shuttle helps to transport this fatty acid into the
 mitochondria. The steps are as follows.
· Acyl group of acyl CoA is transferred to Carnitine, catalyzed by Carnitine acyl
transferase I (present on the outer surface of inner mitochondrial membrane).
· The acyl-carnitine is transported across the membrane to mitochondrial matrix
by a specific carrier protein.
· Carnitine acyl transferase ll (found on the inner surface of inner mitochondrial
membrane) converts acyl-carnitine to acyl CoA.
· The carnitine released returns to cvtosol for reuse

b. How is pyruvate converted in to ethanol in yeast? (2M)

The conversion of pyruvate to ethanol is called alcoholic fermentation and is an

anaerobic pathway of glucose catabolism.

Alcoholic fermentation occurs in two steps:

1. Pyruvate is converted to acetaldehyde by liberation of CO2 in the presence of Pyruvate

decarboxylase enzyme.
2. Acetaldehyde is converted to ethanol by utilizing NADH+H+ in the presence of
enzyme alcohol dehydrogenase.

c. Write the reaction of glycolytic pathway catalysed by Aldolase. (2M)

Aldolase enzyme reversibly catalyses the cleavage of a six carbon Fructose 1,6-bis
phosphate to 2 three carbon compounds namely dihydroxy Acetone phosphate and 3
Phospho glyceraldehyde.
22. a. Why is ATP the most efficient energy molecule? Illustrate with
structure. (4M)

ATP is established as the most efficient energy transfer intermediate occurring in any
cell. It is also called the Universal energy currency. This can be accounted to the large
negative value for standard free energy change of ATP hydrolysis.
ATP + H2O ADP + Pi ; ∆G= -30.5 KJ/mol

The high negative ∆G value is due to the greater stability of ADP when compared to
ATP. The stability of ADP over ATP can be attributed to three main structural
features of ATP:

1. Electrostatic Repulsion-
ATP under normal physiological conditions exists as an anion with four units of
negative charge on the ionised phosphate groups. ADP on the other hand exists as an
anion with three units of negative charge on the ionised phosphate groups. The
localisation of more negative charges induces a greater repulsive force that
destabilises the P-O-P bonds in ATP than ADP.
2. Resonance Stabilization-
The unshared paired of electrons on oxygen is delocalized between successive
phosphorous atoms resulting in resonance stabilized structures in both ATP and
ADP. In addition to ADP on the product side, inorganic phosphate also exhibits
resonance thereby increasing the number of resonance stabilised structures and
hence their stability.
3. Solvation-
Structurally ADP and Pi are smaller when compared to ATP. Hence the products ADP
and Pi are more solvated than ATP. More solvation indicates more stability. Hence
ADP and Pi are more stable than ATP.
 ATP

ADP

b. What is Substrate Level Phosphorylation? Give an example. (2M)

The process where a high energy metabolic intermediate donates the free energy for
driving the synthesis of ATP by phosphorylation of ADP is called Substrate Level
Phosphorylation.

Example:

23. a. Explain Semiconservative mechanism of DNA replication. (4M)

The DNA molecule replicates by Semiconservative mode. The double stranded parent
DNA unwinds to give two separate strands that serve as templates for the synthesis of
new complimentary daughter strands. The parent template and the new daughter
DNA strands recoil to form two DNA molecules identical to the parent.
The semiconservative Replication of DNA involves the following mechanism:

1. Separation of Two complimentary DNA strands.

The DNA replication begins at a unique site called ORI [Origin of Replication].
The ORI site is specifically recognised by the enzyme Unwindase or DNA Helicase.
The enzyme moves along the neighbouring double stranded region to unwind the
DNA double helix.
The separated DNA strands are further bound to SSB [ Single stranded DNA
binding] proteins that prevent the recoiling of the unwound DNA strands.
2. Binding of Gyrase.
The unwinding in the upper end of the helix results in the generation of a negative
pressure at the lower end which might break the molecule. This pressure is relieved
by the enzyme DNA Gyrase which introduces negative supercoiling to facilitate
unwinding.
3. Formation of Replication Fork.
The unwinding and stabilization leads to the formation of a Y shaped structure called
replication fork along which the DNA replication occurs.
4. Initiation of replication.
The DNA polymerase [III] cannot initiate replication independently. They require a
short sequence of RNA at the 3’ end. Such an RNA molecule is called RNA primer and
the enzyme responsible for synthesis of RNA primer is called Primase.
5. Elongation.
The DNA polymerase binds to the RNA primer and synthesises a new DNA strand on
the template DNA strands by adding complimentary nucleotides in the direction 5’ –
3’ direction.
6. On one template strand the synthesis of DNA is continuous. Such a newly synthesised
strand is copied in the direction of advancing replication fork and has its 5’ – 3’
direction towards the replication fork. This strand is called Leading strand.
7. On the other template strand synthesis of DNA is discontinuous. The newly
synthesised strands are copied as fragments in the direction opposing the advancing
replication fork. These fragments are called OKASAKI fragments. This strand
synthesised discontinuously and has its 5’-3’ direction away from the replication fork
is called Lagging strand.
8. To ensure accuracy in replication, the DNA polymerase [III] performs a proof reading
[3’-5’ exonuclease activity].
9. Once the replication fork reaches the end of the double helix, the DNA polymerase [I]
removes the RNA primers by its 5’-3’ exonuclease activity and replaces it with DNA
by 5’-3’ polymerase activity.
10. The fragments of DNA at the end of replication are sealed by DNA ligase to form
continuous DNA strands. These recoil with the template DNA to form a Double
stranded DNA identical to the parent.
 b. What is translation? (2M)

Translation is the process of synthesis of proteins by encoding the information present

on mRNA
It requires an mRNA molecule, Ribosomes, tRNA molecules and activated amino acids.
It occurs in the cytoplasm of cells.

24.a. Mention four salient features of Watson and Crick model of DNA.
(4M)

 It is a double helix with two right handed helical polydeoxy ribonucleotide strands
twisted around the same central axis.
 There are 10 base pair per helix. The pitch of the helix is 36Ao and the distance
between the two strands is 20Ao.
 The two strands are antiparallel. The phosphodiester linkages of one of these strands
runs in 5' to 3' direction while the other strand runs in 3' to 5' direction. The bases are
stacked inside the helix in planes perpendicular to the helical axis.
 The two strands are held together by hydrogen bonds. In addition to hydrogen bonds,
other forces e.g., hydrophobic interactions between stacked bases are also responsible
for stability and maintenance of double helix.
 Adenine always pairs with thymine while guanine pairs with cytosine.
 A-T pair has 2 hydrogen bonds while G-C pair has 3 hydrogen bonds. Hence, G C is
more stronger than A=T.
 The content of adenine is equal to the content of thymine and the content of guanine is
equal to the content of cytosine. This is Chargaff's rule, which is proved by the
complementary base pairing in DNA structure.
 The double helix structure contains major and minor grooves in which proteins
interact with DNA.

b. Write Chargaff’s rule of base equivalance. (2M)

 Chargaff's rules states that DNA from any cell of all organisms should have a 1:1
ratio of pyrimidine and purine bases and the amount of guanine is equal to cytosine
and the amount of adenine is equal to thymine.

25. a. Write a note on Chromatin. (2M)

In the nucleus of each cell, the DNA molecule is packaged into thread-like structures
called chromosomes. Each chromosome is made up of DNA tightly coiled many times
around proteins called histones that support its structure.
The primary functions of chromatin are
1) To pack DNA in to a smaller volume to fit in the cell,
2) To reinforce the DNA macromolecule to allow mitosis
3) To prevent DNA damage, and
4) to regulate gene expression and DNA replication.

b. What is transamination? Give an example. (2M)

The mutual exchange of amino group and keto group between amino acid and α-keto
acid respectively is called transamination. It is an important catabolic reaction of most of
the amino acids. It is catalysed by the enzyme transaminase.

Example:
Alanine transfers amino group to α-ketoglutarate and in turn accepts the keto group to
form Pyruvate. α-ketoglutarate transfers keto group to alanine and in turn accepts the
amino group to form glutamic acid. This reaction is catalyzed by alanine amino
transferase or glutamate-pyruvate transaminase in the presence of pyridoxal
phosphate/PLP.

c. Write any two applications of DNA fingerprinting. (2M)

1. In personal identification: DNA may be the best way to identify a person as all body
tissues and organs contain the same type of DNA.
2. Used for biological evidence to identify criminals for forensic purposes.
3. In solving paternity dispute.
4. To diagnose inherited disorders in both prenatal and new born babies like cystic
fibrosis, hemophilia, sickle cell anemia etc.
5. In the identification of bodies of soldiers killed in war.

[ANY TWO]