BIOCHEMISTRY M.Sc. SEMESTER III (2018-19) UNIT I UNIT II .... III sem.pdf · regulation and...

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BIOCHEMISTRY M.Sc. SEMESTER III (2018-19) 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. Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma Mr. Rohan Gupta Miss Ojal Sohni

Transcript of BIOCHEMISTRY M.Sc. SEMESTER III (2018-19) UNIT I UNIT II .... III sem.pdf · regulation and...

Page 1: BIOCHEMISTRY M.Sc. SEMESTER III (2018-19) UNIT I UNIT II .... III sem.pdf · regulation and energetic of kreb-cycle, Glyoxalate pathway, Pentose phosphate pathway and its physiological

BIOCHEMISTRY M.Sc. SEMESTER III (2018-19)

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. Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma Mr. Rohan Gupta Miss Ojal Sohni

Page 2: BIOCHEMISTRY M.Sc. SEMESTER III (2018-19) UNIT I UNIT II .... III sem.pdf · regulation and energetic of kreb-cycle, Glyoxalate pathway, Pentose phosphate pathway and its physiological

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,

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

Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala

Prof. R.S. Gupta Mr. Deepak Choudhary

Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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BIOCHEMISTRY M.Sc. SEMESTER III (2018-19)

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

Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala

Prof. R.S. Gupta Mr. Deepak Choudhary

Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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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

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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BIOCHEMISTRY M.Sc. SEMESTER III (2018-19)

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. Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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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.

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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BIOCHEMISTRY M.Sc. SEMESTER III (2018-19)

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. Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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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

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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BIOCHEMISTRY M.Sc. SEMESTER III (2018-19)

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

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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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

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

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BIOCHEMISTRY

M.Sc SEMESTER III (2018-19)

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 Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma Mr. Rohan Gupta Miss Ojal Sohni

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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

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma Mr. Rohan Gupta Miss Ojal Sohni

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M.Sc SEMESTER III (2018-19)

Open Elective (Inter- disciplinary)

Paper-: Biochemical Techniques - 4 Credits Unit-I The concept of pH, Dissociation and ionization of acids and bases, pKa buffering mechanism, Henderson-Hasselbalch equation, ionization of amino acids and proteins, measurement of pH. General principle, types of chromatography, adsorption and partition, Column, paper and thin layer. Unit-II Principle, materials used and applications of lon-exchange chromatography, Gel filtration chromatography, Affinity chromatography, Gas Chromatography and High performance liquid chromatography. Unit-III Electrophoresis: General Principle, factors affecting electrophoretic mobility, moving boundary and Zonal Electrophoresis, paper and Gel Electrophoresis, PAGE and SDS PAGE, Isoelectric focusing technique. Sedimentation: Basic principle of sedimentation, Sedimentation velocity, preparative and analytical ultracentrifugation techniques, differential and density gradient centrifugation, Sub cellular fractionation. Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma

Mr. Rohan Gupta Miss Ojal Sohni

Page 14: BIOCHEMISTRY M.Sc. SEMESTER III (2018-19) UNIT I UNIT II .... III sem.pdf · regulation and energetic of kreb-cycle, Glyoxalate pathway, Pentose phosphate pathway and its physiological

Unit-IV Radioactivity: Types of radioisotopes used in Biochemistry (32P,35S,14C and 3H) Disintegration of radio nucleotides, half-life of radioactive compounds, measurement of radioactivity, scintillation counter, Gieger Muller counter, Use of radioisotopes in vivo and in vitro labeling, lsotopic tracer techniques, Autoradiography. Unit-V Spectrophotometry: Beer Lamberts law, extinction coefficient and its importance, design of colorimeter and spectrophotometer. Applications of UV-VIS spectrophotometry, Atomic absorption spectrophotometry. Principle and applications of Optical Rotatory Dispersion, Circular Dichroism, X-ray diffraction and NMR spectroscopy. Reference Books:

1. Principles and techniques of Biochemistry: Wilson and Walker:

2. Biochemistry Laboratory: Modern Theory and Techniques: Rodney Boyer.

3. Biophysical Chemistry (Principles and Techniques) upadhyay and Nath

4. Principles of Biochemistry : A. Lehninger.

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma Mr. Rohan Gupta Miss Ojal Sohni

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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.

Dr. A. Bafna Prof. A. R. Batham Dr. Purnima Dey Sarkar Mrs. Tasneem Rangwala Prof. R.S. Gupta Mr. Deepak Choudhary Prof. R.S. Maheshwari Dr. Bhavna Sharma Mr. Rohan Gupta Miss Ojal Sohni