Syllabus
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Course Code: BCH-203 Course Name: Enzymology |
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| MODULE NO / UNIT | COURSE SYLLABUS CONTENTS OF MODULE | NOTES |
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| 1 | Introduction: Historical perspectives; General characteristics; Nomenclature and classification; Introduction to the following terms with examples – Holoenzyme, apoenzyme, cofactors, coenzymes, prosthetic groups, metalloenzymes, turnover number, enzyme activity units (I.U and Katal), and specific activity.Multienzyme systems and multifunctional enzymes with specific examples and significance. Enzyme specificity: Types of specificity; three-point attachment theory to explain stereospecificity; Lock-and-key hypothesis; Induced- fit hypothesis; Hypothesis involving strain or transition-state stabilization.Enzyme Catalysis: Role of NAD+/NADP+, FMN/FAD, coenzyme A, thiamine pyrophosphate, pyridoxal phosphate, lipoic acid, biocytin, Vitamin B12 Coenzyme, and tetrahydrofolate coenzymes in enzyme catalysis; Common features of active sites;Reactionco-ordinate diagram; Proximity & orientation, acid-base catalysis, and covalent catalysis; Mechanism of action of chymotrypsin, ribonuclease, carboxypeptidase, and lysozyme |
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| 2 | Enzyme assay: Introduction; Kinetic and coupled enzyme assays. Enzyme Kinetics: Factors affecting enzyme activity; Arrhenius plot; Derivation of Michaelis-Menten equation for unisubstrate reactions; Km and its significance; Kcat/Km and its importance; Measurement of Km and Vmax by Lineweaver-Burk plot and other linear transformations of MM equation; Bi-substrate reactions: Sequential and ping-pong mechanisms with examples and determination of Km and Vmax for each substrate (derivations excluded); Use of initial velocity studies, product-inhibition studies and isotope exchange at equilibrium for determining the kinetic mechanism of a bisubstrate reaction. |
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| 3 | Methods of studying fast reactions: A brief account of rapid mixing techniques, flash photolysis and relaxation methods. Enzyme inhibition: Reversible (competitive, non- competitive, and uncompetitive) and irreversible (affinity labels and suicide inhibitors) enzyme inhibitors; Determination of Ki. Investigation of active site structure: Methods for identification of binding and catalytic sites- Trapping the enzyme-substrate complex, use of substrate analogues, chemical modification of amino acid side chains in enzymes, enzyme modification by proteases and effect of changing pH. |
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| 4 | Enzyme regulation: Coarse and fine control of enzyme activity; Enzyme induction & Repression; Feedback inhibition; Allosteric enzymes with aspartate transcarbamoylase as an example; Concerted and sequential models for action of allosteric enzymes; Negative and Positive Cooperativity; Hill plot; Scatchard plot; Regulation by reversible and irreversible covalent modification of enzymes; Isoenzymes. Ribozyme and Abzyme |
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