Enzymology

Introduction to enzymology, nomenclature and classification of enzymes, properties of enzymes, enzyme assay and units of activity, isolation and purification of enzymes, Concept of structure and nature of active site, stereospecificity and ES complex formation. Activation energy. Transition state theory. Enzyme activity, international units, specific activity, turnover number. Catalytic RNA.

Acid-base, covalent, proximity and orientation, transition state binding and metal ion effects of enzyme catalysis, metal ion and electrostatic catalysis.

Chemical kinetics, Michaelis-Menten and Briggs-Haldane kinetics.Determination of Km analysis of kinetic data, Double Recipocal plot.  importance of Km, Ki and Vmax ,half life.
Allosteric enzymes, sigmoidal kinetics and their physiological significance, symmetric and sequential modes for action of allosteric enzymes and their significance. Hill and Scatchard plots and kinetics of allosteric enzymes. Cooperativity, positive and negative cooperativity, oxygen binding to hemoglobin, Hill equation homotropic and heterotropic effectors, aspartyl transcarbamylase as an allosteric enzyme.
 

Effect of temperature, pH and substrate concentration on reaction rate. Product inhibition, feedback control, enzyme induction and repression and covalent modification. isozymes and zymogens, enzyme inhibitors, Enzyme inhibition - types of inhibitors - competitive, noncompetitive and uncompetitive, their mode of action and experimental determination. feed-back inhibition and repression.

Various methods of immobilization - ionic bonding, adsorption, covalent bonding (based on R groups of amino acids) , microencapsulation and gel entrapment.  Immobilized multienzyme systems
Relative practical and economic advantage for industrial use, effect of partition on kinetics and performance with particular emphasis on charge and hydrophobicity (pH, temperature and Km).