Enzyme Technology Fourth Semester Syllabus for BSc Biotechnology and BSc Microbiology
Unit I- 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
Unit II- 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-Chymotryspin, 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 Inhibitions like Penicillin, Iodoacetamide and DIPF.
Unit IV 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. Kinetics Allosteric enzymes.
Unit III- Enzyme Regulation
Feedback Regulation, Allosteric Regulation, Reversible Covalent Modification and Proteolytic Activation. Organisation of enzymes in the cell. Enzymes in the cell, localization, compartmentation of metabolic pathways, enzymes in membranes, concentrations. Mechanisms of enzyme degradation, lysosomal and nonlysosomal pathways, examples.
Unit V 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-inflamatory agents, strptokinasae, 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 VI 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.
- Fundamentals of Enzymology : Nicholas Price & Lewis Stevens
- Enzymes : Biochemistry, Biotechnology and Clinical Chemistry- Trevor Palmer
- Biochemistry text books by Stryer, Voet and Lehninger (Relevant Chapters)
- Proteins by Gary Walsh
- Internet/ Journal Resources