Syllabus
Course Code: IN-BTY-201 Course Name: Enzymology |
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MODULE NO / UNIT | COURSE SYLLABUS CONTENTS OF MODULE | NOTES |
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1 | Enzymes: Historical perspectives, general characteristics, nomenclature & classification, significance of numbering system, holoenzyme, apoenzyme, coenzymes, cofactors, activators, inhibitors, active site, metallo-enzymes, isoenzymes, monomeric enzymes, oligomeric enzymes, multifunctional enzyme and multi-enzyme complexes. Enzyme specificity. Measurement and expression of enzyme activity: Enzyme assay, enzyme units, enzyme turn over number and specific activity. Role of cofactors in enzyme catalysis: NAD/NADP, FMN/FAD, coenzyme A, biocytin, Vitamin B12 Coenzyme, lipoamide, TPP, pyridoxal phosphate, tetrahydrofolate and metal ions with special emphasis on coenzyme functions Enzyme catalysis: Reaction co-ordinate diagram, transition state, Acid-base catalysis, covalent catalysis, proximity and orientation effects, strain and distortion theory. Mechanism of action of chymotrypsin, carboxypeptidase, andribonuclease. Enzyme Purification: Methods of isolation of enzymes, purification of enzymes - ammonium sulfate precipitation, molecular-sieving, ion-exchange chromatography, affinity chromatography, criteria of homogeneity and determination of molecular weight of enzyme. |
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2 | Enzyme Kinetics: Factors affecting enzyme activity- enzyme concentration, substrate concentration, pH and temperature. Derivation of Michaelis - Menten equation for uni-substrate reactions. Km and its significance.Lineweaver-Burk plot.Importance of Kcat/Km. Bi-substrate reactions- brief introduction of sequential and ping-pong mechanisms with examples.Reversible (competitive, non-competitive and uncompetitive inhibitions) and irreversible inhibition.Determination of Km &Vmax in the presence and absence of inhibitor. Enzyme regulation: Feedback inhibition, Allosteric enzymes. Covalently modulated enzymes.Zymogen activation. Immobilized enzymes: Advantages, methods of immobilization - Adsorption, ionic binding, covalent coupling, cross-linking, entrapment, microencapsulation etc. Applications of immobilized enzymes (A brief account).Industrial applications of enzymes (Production of glucose from starch, cellulose and dextran; use of lactase in dairy industry; production of glucose-fructose syrup from sucrose; use of protease in food, detergent and leather industry). |