Enzyme Engineering

Holoenzyme, apoenzyme, prosthetic group. Interaction between enzyme and substratelock and key model, induced fit model. Features of active site, activation energy, enzyme specificity and types. IUB system of classification and nomenclature of enzymes. Kinetics of single substrate reactions; Derivation of Michaelis - Menten equation, turnover number; determination of Km and Vmax (LB plot, ED plot), Importance of Km &Vmax; Numerical related to enzyme kinetics, Multi-Substrate reaction mechanisms.

Factors affecting the velocity of enzyme catalyzed reaction- enzyme concentration, temperature, pH, substrate concentration, inhibitors and activators. Enzyme inhibition: irreversible; reversible (competitive, uncompetitive and non competitive inhibition); Substrate and Product inhibition, Allosteric regulation of enzymes, concerted & sequential model; Deactivation Kinetics.

Extraction of crude enzyme from plant, animal and microbial source; some case study. Purification of enzymes by the help of different methods. Methods of characterization of enzymes; criteria of purity. Unit of enzyme activity - definition and importance. Development of enzyme assays.

Adsorption, Matrix entrapment, Encapsulation, Cross linking, Covalent binding and their examples; Advantages and disadvantages of different immobilization techniques. Structure & stability of immobilized enzymes, kinetic properties of immobilized enzymes- partition effect, diffusion effect. Overview of applications of immobilized enzyme systems.

elements of biosensors, three generations of biosensors, Types of biosensors: calorimetric, potentiometric, amperometric, optical and piezoelectric. Design of enzyme electrodes and their applications as biosensors in industry, health care and environment. Design of Immobilized Enzyme Reactors- Stirred tank reactors(STR), Continuous Flow Stirred Tank Reactors (CSTR), Packed- bed reactors (PBR), Fluidized-bed Reactors (FBR); Membrane reactors.