Course

Unit 1

Introduction to Enzymes: General introduction and historic background- General Terminology, Nomenclature and Classification of Enzymes. Criteria of purity of enzymes- Specific activity. Enzyme units-Katal and IU. Enzyme activity- chemical nature of enzymes. Protein nature of enzymes and Non protein enzymes- Ribozymes and DNAzymes. Metalloenzymes and metal activated enzymes. Coenzymes and Cofactors- Prosthetic group, coenzymes involved in different metabolic pathways. Classification of coenzymes. Isozymes, Abzymes, Synzyme, Specificity of enzymes, Active site, Allosteric site.

Unit 2

Enzyme Catalysis and Inhibition: Lock and key, Induced fit and Transition state Hypotheses. Mechanism of enzyme catalysis- Acid-base catalysis, covalent catalysis, Metal ion catalysis, Proximity and orientation effects etc. mechanism of Serine Proteases-Chymotrypsin, Lysozyme, Carboxypeptidase A and Ribonuclease., Proenzymes (Zymogens). Reversible Inhibition- Competitive, Non-Competitive, Uncompetitive, Mixed, Substrate, Allosteric and Product Inhibition. Irreversible Inhibition- Suicide inhibition. Examples and Mechanism of various Inhibitors like Penicillin, Iodoacetamide and DIPF.

Unit 3

Enzyme Kinetics: Factors affecting the enzyme activity- Concentration, pH and temperature. Kinetics of a single-substrate enzyme catalysed reaction, Michealis-Menten Equation, Km, Vmax, L.B Plot, Turnover number, Kcat. Kinetics of Enzyme Inhibition. Kinetic Allosteric enzymes. Numerical problems in enzyme kinetics and enzyme inhibition.

Unit 4

Enzyme Regulation: Feedback Regulation, Allosteric Regulation, Reversible, Covalent Modification and Proteolytic Activation. Enzyme processing. Enzymes in post translational modifications.Organization of enzymes in the cell. Enzymes in the cell, localization, compartmentation of metabolic pathways, enzymes in membranes, concentrations. Mechanisms of enzyme degradation, lysosomal and non-lysosomal pathways, examples.

Unit 5

Industrial and Clinical uses of Enzymes (Applied Enzymology): Industrial Enzymes- Thermophilic enzymes, amylases, lipases, proteolytic enzymes in meat and leather industry, enzymes used in various fermentation processes, cellulose degrading enzymes, Metal degrading enzymes.Clinical enzymes- Enzymes as thrombolytic agents, Anti-inflammatory agents, streptokinase, asparaginase, Isoenzymes like CK and LDH, Transaminases (AST, ALT), Amylases, Cholinesterases, Phosphatases. Immobilization of enzymes, ELIZA. Biosensors. Enzyme Engineering and site directed mutagenesis, Designer enzymes

Unit 6

Enzyme Structure activity Relationship (SAR) and Drug Discovery- Properties of Enzymes.: Lead Compound, Structure based drug design, combinatorial chemistry, High-throughput screening, Case study of DHFR etc.

LEARNING OBJECTIVES:

To provide a detailed knowledge about enzymes, their chemical nature, kinetics, catalysis, classifications, factors affecting the velocity of enzymes, theories of enzyme action, enzyme regulation, inhibitions, clinical enzymes, industrial enzymes, non-protein enzymes, coenzymes and cofactors.

COURSE OUTCOMES:

After completing the course, students shall be able to

CO1. Understand and define the basic concepts of enzymes, their classification, metalloenzymes, holoenzymes, abzymes, Isozymes, Multienzyme complex.

CO2. Differentiate and discuss the different catalytic mechanisms with examples, transition state theory, Lock and key and induced fit theories. Zymogens, Proenzymes.

CO3. Discuss and explain enzyme kinetics, Michaelis Menton equation, LB plot, Km, Vmax, Kcat and Turnover number. Enzyme inhibitions: reversible and irreversible with example; Kinetics of inhibitions; Allosteric enzymes: regulation of enzymes

CO4. Demonstrate and explain the role of enzymes in industry and medicine with examples.

REFERENCES:1. Nicholas Price & Lewis Stevens, Fundamentals of Enzymology, 3rd Edition; 2009. 2. Trevor Palmer and P L Bonner, Enzymes: Biochemistry, Biotechnology and Clinical Chemistry, 2nd Edition, 2007.3. Lehninger, Nelson and Cox, Principles of Biochemistry, 7th Edition, 2016.4. Donald Voet, Judith G. Voet, Charlotte W. Pratt, Fundamentals of Biochemistry: Life at the Molecular Level, 5th Edition. 2016.5. Gary walsh, Proteins, 2nd Edition, 2015.6. Internet/Journal Resources