Micro-Credential in Enzyme and Protein Purification Technology

Week 1

Introduction to enzymes

·         Classification – IUB system

·         Rationale, overview and specific examples

·         Characteristics of enzymes, enzyme substrate complex

·         Concept of active center, binding sites, stereo specificity and ES complex formation

·         Effect of temperature, pH and substrate concentration on reaction rate

·         Activation energy

·         Transition state theory Enzyme activity

·         International units

·         Specific activity

·         Turnover number

·         End point kinetic assay

Lecture

Week 2

Enzyme kinetics:

·         Michaelis – Menten Equation – form and derivation

·         Steady state enzyme kinetics

·         Significance of Vmax and Km

·         Bisubstrate reactions

·         Graphical procedures in enzymology

·         Advantages and disadvantages of alternate plotting

·         Enzyme inhibition – types of inhibitors – competitive, noncompetitive and uncompetitive, their mode of action and experimental determination

Lecture, tutorial

Week 3

Enzyme catalysis

·         Factors affecting catalytic efficiency

·         Proximity and orientation effects

·         Distortion or strain

·         Acid – base and nucleophilic catalysis

·         Methods for studying fast reactions

·         Chemical modification of enzymes

·         Isoenzymes and multiple forms of enzymes

Lecture, practical

Week 4

Enzyme regulation:

·         Product inhibition

·         Feedback control

·         Enzyme induction and repression and covalent modification

·         Allosteric regulation

Lecture

Week 5

Immobilization of enzymes:

·         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)

·         Various methods of immobilization- ionic bonding, adsorption, covalent bonding (based on R groups of amino acids), microencapsulation and gel entrapment.

·         Immobilized multi-enzyme systems

Lecture, practical

Week 6

Current applications of enzymes:

·         Food

·         Agriculture

·         Medicine

·         Environment

·         Industry

Protein chemistry and structure determination:

·         Introduction and brief review on level of protein structure

·         Properties & classification

·         Structure determination by X-ray crystallography and NMR spectroscopy

·         Structure model and analysis using molecular graphing

Protein Folding:

·         Basic concepts

·         Protein folding and diseases

·         Protein-folding models

·         Experimental approaches

·         Crystallographic determination of protein structure

Lecture, practical

Week 7

Protein Purification (Micro to Macro):

·         Classical and modern techniques in protein purification

·         Micro purification and analysis

·         Purification of synthetic proteins

·         Bulk precipitation and phase partition methods

·         Chromatographic techniques in processing scale

·         FPLC system

·         New separation concepts

Lecture, tutorial, practical

Week 8

Final Examination

Conclusion Session

Awarding of Certificates

Presentation by VC or Representative of VC