Biochemical Engineering

Production and Purification of Enzymes:

Kinetics and Mechanism of Enzyme Action:

Mechanism of Enzyme Action:

An enzyme attracts substrates to its active site, catalyzes the chemical reaction by which
products are formed, and then allows the products to dissociate (separate from the enzyme
surface). The combination formed by an enzyme and its substrates is called the enzyme–
substrate complex. When two substrates and one enzyme are involved, the complex is called a
ternary complex; one substrate and one enzyme are called a binary complex. The substrates are
attracted to the active site by electrostatic and hydrophobic forces, which are called
noncovalent bonds because they are physical attractions and not chemical bonds.
As an example, assume two substrates (S1 and S2) bind to the active site of the enzyme during
step 1 and react to form products (P1 and P2) during step 2. The products dissociate from the
enzyme surface in step 3, releasing the enzyme. The enzyme, unchanged by the reaction, is able
to react with additional substrate molecules in this manner many times per second to form
products. The step in which the actual chemical transformation occurs is of great interest, and,
although much is known about it, it is not yet fully understood. In general there are two types
of enzymatic mechanisms, one in which a so-called covalent intermediate forms and one in
which none forms.

Kinetics of Enzyme Reaction:

Enzyme kinetics is the study of the chemical reactions that are catalysed by enzymes .In enzyme
kinetics, the reaction rate is measured and the effects of varying the conditions of the reaction
are investigated. Studying an enzyme's kinetics in this way can reveal the catalytic mechanism
of this enzyme, its role in metabolism, how its activity is controlled, and how a drug or an
agonist might inhibit the enzyme.

Enzyme kinetics, which investigates the rates of enzyme-catalyzed reactions as affected by

various factors, offers an enormous potential to the study of enzyme reaction mechanisms and
functions. Some important factors that affect the rates of enzymatic reactions are enzyme
concentration, ligand (substrates, products, inhibitors, and activators) concentrations, solvent
(solution, ionic strength, and pH), and temperature. When all these factors are properly
analyzed, it is possible to learn a great deal about the nature of enzymes.

Michaelis-Menten kinetics:

Kinetics of simple enzyme catalyzed reactions are often referred to as Michaelis- Menten
Kinetics or saturation kinetics. The model takes the form of an equation describing the rate
of enzymatic reactions, by relating reaction rate to substrate concentration or a system where a
substrate S binds reversibly to an enzyme E to form an enzyme-substrate complex ES, which
then reacts irreversibly to generate a product P and to regenerate the free enzyme E. This
system can be represented schematically as follows: