biochemistry m.sc. semester iii (2017-18) unit i … iii sem paper… · ·...

BIOCHEMISTRY

M.Sc. SEMESTER III (2017-18)

Paper 1: Enzymology (Core) - 4 Credits

UNIT – I

Isolation and purification, Classification and nomenclature of enzyme .

Enzyme catalysis: Enzyme specificity and the concept of active site , determination

of active site. Stereospecificity of Enzymes.

UNIT – II

Enzyme kinetics: Factors affecting rates of enzymes catalyzed reaction ,

unisubstrate reaction , concept of Michaelis – Menten, Briggs - Haldane

relationship . Determination and significance of kinetic constants , catalytic rate

constant and specificity constant . Limitations of Michaelis – Menten kinetics.Line

weaver Burk plot and its limitation.

UNIT – III

Classification and kinetics of multisubstrate reaction, sequential and ping pong bi

bi models with examples.

Reversible and irreversible inhibition, competitive, non competitive and

uncompetitive inhibitions.

UNIT – IV

Mechanism of catalysis: Proximity and orientation effects, general acid – base

catalysis, concerted acid – base catalysis, nucleophilic and electrophilic attacks,

catalysis by distortion, metal ion-cataysis. Theories on mechanism of catalysis.

Mechanism of enzyme action: mechanism of action of Lysozyme, Chymotrypsin,

Carboxypeptidase. Multienzyme system, Mechanism of action, regulation and

coenzyme of Pyruvate Dehydrogenase and Fatty acid Synthetase complexes.

Prof. A.R. Batham Dr. A. Bafna

Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala

Prof. R.S. Gupta Mr. Deepak Choudhary

Prof. R.S. Maheshwari Mrs Surya Sugandhi

Mr. Rohan Gupta Miss Pooja Gupta

UNIT – V

Enzyme regulation: General mechanisms of enzyme regulation, Allosteric

enzymes, sigmoidal kinetics and their physiological significance. Symmetric and

sequential modes for action of allosteric enzymes. Reversible and irreversible

covalent modification of enzymes.

Immobilized enzymes and their industrial applications.

REFERENCE BOOKS-

1. Understanding Enzymes by T. Palmer

2. Enzymes by M. Dixon and Ec. Webb

3. The Enzymes by PD Boyer

4. Principles of Biochemistry by Geoffrey L Zubey,






BIOCHEMISTRY


Paper 2: Metabolism I (Core) 4 Credits

UNIT – I

The concept of Gibbs free energy, exergonic and endergonic reactions , redox

potential. High energy bond and key position of ATP, Substrate level and

oxidative phosphorylation. Electron Tansport Chain, Inhibitors and uncouplers of

Electron Tansport Chain.

UNIT – II

Carbohydrates: Glycolysis and glycogenolysis and their regulation, glycogen

storage diseases Glycogenesis and Gluconeogenesis, Cori’s cycle. Role of

hormones in blood sugar regulation.

UNIT – III

Degradation of carbohydrates: Pyruvate dehydrogenase complex, reactions,

regulation and energetic of kreb-cycle, Glyoxalate pathway, Pentose phosphate

pathway and its physiological significance, Uronic acid pathway and its

importance.

UNIT – IV

Lipids: Oxidation of fatty acids- mitochondrial β-Oxidation, α-Oxidation and ω-

Oxidation, Oxidation of unsaturated and odd-chain fatty acids, ketone bodies.

Biosynthesis of fatty acid- saturated and unsaturated.






UNIT – V

Phospholipids and glycosphingolipids- synthesis and degradation, lipid storage

diseases.

Salient features of the metabolism of cholesterol and bile acids.

Regulation of lipid metabolism.

REFERENCE BOOKS-

1. Textbook of Biochemistry by West Todd

2. Biochemistry by A. Lehninger

3. Review of Biochemistry by Harper

4. Biochemistry by Stryer Lubert






BIOCHEMISTRY


Paper 3: Plant Biochemistry (Elective) - 4 Credits

UNIT – I

Structure of Chloroplast, Photosynthesis, dessipation of excitation energy by

chlorophyll, photosynthetic electron transport chain. Thylakoid membrane protein

complexes – PS I, PS II, LHC II, Cyt b6f, ATP synthase complexes, cyclic

photophosphorylation.

UNIT – II

Calvin cycle: Biochemistry of RuBP Carboxylase/oxygenase, activation of

RUBISCO, oxygenation reaction, photorespiration and compensation point,

photosynthetic efficiency. Regulation of enzymes of carbon dioxide fixation by

light;Difference between C3 and C4 plants Hatch and slack pathway, CAM plants.

UNIT – III

Nitrogen metabolism: Nitrogen fixation, nitrogenase complex, mechanism of

action of Nitrogenase. Structure of ‘NIF’ genes and its regulation. Hydrogen

uptake and bacterial hydrogenases.

Nitrate metabolism: Enzymes of nitrate metabolism, regulation of their synthesis

and activity.

UNIT – IV

Ammonia assimilation, Nitrification, Denitrification, Nitrate and sulphate

reduction and their incorporation in amino acids.

Translocation of organic and inorganic substances.






UNIT – V

Special features of secondary plant metabolism, formation of phenolisc, tannins,

lignin, terpenes, terpenoids and alkaloids.

Water and mineral balance in plants.

Structure, function and mechanism of action of phytohormones- Auxin Gibberlin

ABA, Cytokinin and Ethylene.

Defence system in plants.

REFERENCE BOOKS-

1. Plant biochemistry by P.M. Dey & J.B. Harborane

2. Plant biochemistry by Geza Doby

3. Plant biochemistry by Bonner and Varner

4. Recent Advances in Plant biochemistry by Mehta S.L. and Lodha M.L.






BIOCHEMISTRY


Paper 4: Biotechnology (Elective) - 4 Credits

UNIT – I

Basic principles of genetic engineering: Methods of creating recombinant DNA

molecule, splicing, properties of restriction endonucleases and their mode of

action. Construction of DNA library: Genomic library and cDNA library, chemical

synthesis of gene.

UNIT – II

Structure, selection and screening of Cloning vectors (lambda phage, plasmid, M-

13 phage, cosmid, shuttle vectors) and expression vectors

UNIT – III

Analysis of genomic DNA by southern hybridization, Northern and Western

blotting techniques.

Restriction mapping: Restriction fragment length polymorphism (RFLP), Rapid

amplified polymorphic DNA (RAPD) and Amplified fragment length

polymorphism (AFLP).

UNIT – IV

DNA sequencing techniques: plus and minus, dideoxynucleotide, Maxam and

Gilbert method, Amplification of DNA by polymerase chain reaction

(PCR).Appliocations and variation of PCR.

Site directed mutagenesis. Antisense RNA technique.






UNIT – V

Transfection methods for animals and plants electroporation, particle gun,

microinjection.

Transformation techniques for plants-: Agrobacterium mediated gene transfer and

direct gene transfer.

Application of genetic engineering in medicine and agriculture, vaccine

production.

Reference Books-

1. Gene Cloning An Introduction by T.A. Brown

2. Principles of Gene Manipulation by R.W. Old and Primrose

3. Biotechnology by B.D. Singh

4. Elements of Biotechnology by P.K. Gupta






BIOCHEMISTRY


Paper 5: Environmental Toxicology (Elective) - 4 Credits

UNIT – I

Fundamental of toxicology.Definition and Scope.Types of toxic substances.

Dose response relationship.Xenobiotic metabolism: 1) Absorption 2) Distribution

3) Metabolism 4) Phase –I reaction and Phase II reaction.

UNIT – II

Types of exposure.Types of toxic response.

Tissue specificity and organ specificity of toxicity (w.r.t. liver, lungs, kidney)

Diagnosis of toxic changes in liver and kidney.Drugs as toxic substances.

Metabolism of haloalkanes,haloalkenes and paracetamol with their toxic effects on

tissue.

Food toxicology: Role of diet in cardio vascular disease and cancer, Toxicology of

food additives.

UNIT – III

Pesticide’s Toxicology: 1) Insecticide toxicology: Organochlorines,

Organophosphates, Carbamates 2) Herbicide toxicology Paraquat.

Metal toxicity: 1) Arsenic 2) Mercury 3) Lead 4) Cadmium






UNIT – IV

Environmental Pollution: Air Pollution, Common air pollutants and their sources,

acid rain, ozone layer depletion, water pollution.

UNIT – V

Toxicity by natural products: Plant toxins, Animal Toxins, Microbial Toxins

(Fungal and bacterial)

Toxicity by household products: carbon monoxide, Antifreeze, Ethylene glycol,

Alcohol.

Toxicity testing: Acute Toxicity Test, Sub Acute Toxicity Tests, Chronic Toxicity

Tests

REFERENCE BOOKS-

1. Principles of Biochemical Toxicology by John Trimbell

2. Introduction to Toxicology by J.A. Trimbell

3. Pharmacology and Toxicology by B. K. Rao

4. Textbook of veterinary Toxicology by H.S. Sandhu, R.S. Brar






BIOCHEMISTRY

M.Sc SEMESTER III (2017-18)

Paper-6 Advanced Biochemistry (Elective) - 4 Credits

Unit-I – Genome Organization

Organization & Structure of eukaryotic chromosome

Role of nuclear matrix in chromosome organization & function

Heterochromatin & Euchromatin

DNA reassociation kinetics (Cot Curve analysis), repetitive & unique sequence

Satellite DNA, DNA melting & buoyant density, nucleoside phasing

Unit-II- Molecular Biological Technique

Nucleic acid isolation – Isolation of genomic DNA, RNA, Plasmid DNA

PCR, Blotting techniques & hybridization technique

RNA Interference

Genetic mapping & genetic marker – RFLP, mini & macro satellite, RAPD, AFLP

Gene knockout

Unit-III- Cell Signaling

Signaling molecules & their receptors

Function of G-Protein- coupled receptor

Protein tyrosine kinase & cytokine receptor

Pathway of intracellular signal transduction






Unit-IV- Plant Metabolism

Photosynthesis-general features of photophosphorylation, Light absorption, light-

driven electron flow, ATP synthesis by phosphorylation.

N 2 metabolism- Biological N2 fixation, nitrogenase complex, biotechnology of N2

fixation, role of leghemoglobin, regulation of nif gene

Unit-V- Metabolic disorder

Inborn error of protein metabolism

Inborn error of carbohydrate metabolism

Inborn error of lipid metabolism, lipid storage diseases.

Diabetes, Atherosclerosis, Arthritis, Gout.

REFERENCE BOOKS-

1. Principles of Gene Manipulation by R.W. Old and Primrose

2. Cell Biology by Karp

3. Recent Advances in Plant biochemistry by Mehta S.L. and Lodha M.L.

4. Biochemical Disorders in Human Disease by Thompson and Wootton

5. Biotechnology by B.D. Singh






M.Sc. Semester III Practical 1

Practical List

1. Extraction and estimation of Proteins from plant material.

2. Extraction and estimation of RNA from plant material.

3. Extraction and estimation of DNA from plant material.

4. Estimation of chlorophyll and carotenoid pigments in plant material.

5. Determination of peroxidase activity in plant material.

6. Determination of Nitrate reductase activity in plant material.

7. Determination of Glutamate dehydrogenase activity in plant material.

M.Sc. Semester III Practical 2

Practical List

1. Estimation of activity of salivary amylase by DNS method.

2. Effect of substrate concentration on enzyme activity.

3. Effect of enzyme concentration on enzyme activity.

4. Effect of temperature on enzyme activity

5. Effect of pH on enzyme activity.






biochemistry m.sc. semester iii (2017-18) unit i … iii sem paper… · ·...

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