BT3452 - Industrial Enzymology
Comprehensive Notes for Students
1. Introduction to Enzymes
Enzymes are biological catalysts that accelerate chemical reactions in living organisms.
They play a crucial role in industrial applications such as pharmaceuticals, food processing,
and textiles.
1.1 Classification of Enzymes
Enzymes are classified into six major categories based on the type of reaction they catalyze:
1. Oxidoreductases – Involved in oxidation-reduction reactions.
2. Transferases – Transfer functional groups between molecules.
3. Hydrolases – Catalyze hydrolysis reactions.
4. Lyases – Add or remove groups to form double bonds.
5. Isomerases – Convert molecules into their isomeric forms.
6. Ligases – Join two molecules using ATP.
1.2 Mechanism of Enzyme Action
Enzymes lower the activation energy of biochemical reactions by stabilizing the transition
state. The enzyme-substrate interaction follows the 'lock and key' model or the 'induced fit'
model.
1.3 Active Site and Enzyme Specificity
The active site of an enzyme is the region where the substrate binds. Enzyme specificity is
classified as:
- Absolute specificity: Acts only on a single substrate.
- Group specificity: Acts on molecules with a specific functional group.
- Bond specificity: Recognizes specific chemical bonds.
- Stereochemical specificity: Acts on a particular enantiomer.
2. Enzyme Kinetics
Enzyme kinetics is the study of the rates of enzyme-catalyzed reactions. The Michaelis-
Menten equation describes the relationship between substrate concentration and reaction
velocity.
2.1 Michaelis-Menten Equation
The Michaelis-Menten equation is:
v = (Vmax [S]) / (Km + [S])
where:
- v is the reaction rate
- Vmax is the maximum reaction rate
- [S] is the substrate concentration
- Km is the Michaelis constant
2.2 Enzyme Inhibition
Enzyme inhibitors slow down enzyme activity. Types of inhibition include:
- Competitive inhibition: Inhibitor competes with substrate for active site.
- Non-competitive inhibition: Inhibitor binds to an allosteric site.
- Uncompetitive inhibition: Inhibitor binds only to the enzyme-substrate complex.
3. Production and Purification of Enzymes
Industrial enzymes are produced using microorganisms such as bacteria, fungi, and yeast.
Fermentation techniques include:
- Submerged fermentation (SmF)
- Solid-state fermentation (SSF)
3.1 Enzyme Purification
Steps involved in enzyme purification:
1. Cell disruption (mechanical or chemical methods)
2. Precipitation (using ammonium sulfate)
3. Chromatography (ion-exchange, affinity, gel filtration)
4. Electrophoresis (SDS-PAGE for molecular weight determination)
4. Immobilized Enzyme Technology
Immobilized enzymes offer advantages such as reusability, stability, and ease of separation.
Methods of immobilization include:
- Adsorption
- Covalent bonding
- Entrapment
- Encapsulation
4.1 Kinetics of Immobilized Enzymes
Mass transfer limitations affect the activity of immobilized enzymes. The diffusion of
substrate through the immobilization matrix is an important factor in industrial
applications.
5. Enzyme Applications
Enzymes are widely used in industries such as:
- Food industry: Amylases for starch processing, proteases for meat tenderization
- Pharmaceutical industry: Penicillin amidase for antibiotic production
- Textile industry: Cellulases for fabric softening
- Clinical diagnostics: Enzymes as biosensors for glucose detection
5.1 Enzymes in Biotransformations
Enzymes catalyze the transformation of substrates into valuable products, such as:
- High-fructose corn syrup production (glucose isomerase)
- Semi-synthetic penicillins (penicillin acylase)
- Biofuel production (lipases for biodiesel synthesis)
Conclusion
Industrial enzymology plays a key role in biotechnology, pharmaceuticals, food, and textile
industries. Understanding enzyme kinetics, production, purification, and applications is
essential for biotechnologists.
