Production of Enzymes — General Considerations and Examples

General Considerations in Industrial Enzyme Production

1. Selection of Microorganisms

   • Enzyme-producing bacteria (Bacillus), fungi (Aspergillus spp.), and yeast (Saccharomyces spp.).

2. Fermentation Process

   • Submerged Fermentation (SmF): Microorganisms are grown in liquid media (e.g., for amylase production).
   • Solid-State Fermentation (SSF): Uses solid substrates like bran or wheat for enzyme production (e.g., fungal proteases).

3. Optimization of Growth Conditions

   • pH, temperature, aeration, and nutrient supply are adjusted for maximum enzyme yield.

4. Downstream Processing

   • Includes enzyme extraction, purification, and formulation.

Production of Specific Enzymes

Amylase

• Function:

  • Hydrolyzes starch into simpler sugars (maltose, glucose).

• Sources:

  • Bacillus subtilis
  • Aspergillus niger
  • Bacillus amyloliquefaciens

• Production Method:

  • Fermentation: Produced via submerged fermentation using starch as a substrate.
  • Purification: Precipitation, dialysis, and chromatographic techniques.

• Applications:

  • Food industry (bread, beer)
  • Textile industry (desizing fabrics)
  • Detergent formulations

Catalase

1. Function:

   • Breaks down hydrogen peroxide (H₂O₂) into water and oxygen.

2. Sources:

   • Micrococcus luteus
   • Aspergillus niger
   • Bacillus species

3. Production Method:

   • Fermentation: Typically grown in glucose-rich media under aerobic conditions.
   • Purification: Filtration, precipitation, ion-exchange chromatography.

4. Applications:

   • Textile industry (removal of H₂O₂ from fabrics)
   • Food processing
   • Medical applications (wound cleaning)

Peroxidase

1. Function:

   • Catalyzes oxidation reactions using hydrogen peroxide as an oxidant.

2. Sources:

   • Horseradish (HRP)
   • Aspergillus niger
   • Bacillus

3. Production Method:

   • Fermentation: Cultivation of fungal or bacterial strains in optimized media.
   • Purification: Filtration, affinity chromatography.

4. Applications:

   • Diagnostic kits
   • Wastewater treatment
   • Biosensors and immunoassays

Lipase

1. Function:

   • Hydrolyzes triglycerides into glycerol and fatty acids.

2. Sources:

   • Candida rugosa
   • Pseudomonas aeruginosa
   • Aspergillus

3. Production Method:

   • Fermentation: Produced via submerged or solid-state fermentation using oils and fats as inducers.
   • Purification: Precipitation, ultrafiltration, chromatography.

4. Applications:

   • Detergents
   • Dairy industry (cheese, butter)
   • Pharmaceuticals (digestive aids)

Protease

1. Function:

   • Hydrolyzes proteins into peptides and amino acids.

2. Sources:

   • Bacillus subtilis
   • Aspergillus oryzae
   • Streptomyces griseus

3. Production Method:

   • Fermentation: Typically produced in alkaline or neutral media using submerged fermentation.
   • Purification: Ultrafiltration, precipitation, chromatography.

4. Applications:

   • Detergents
   • Leather industry (dehairing hides)
   • Pharmaceuticals (digestive enzymes)

Penicillinase (Beta-lactamase)

• Function:

  • Breaks down penicillin, conferring resistance to β-lactam antibiotics.

• Sources:

  • Bacillus cereus
  • Staphylococcus aureus

• Production Method:

  • Isolation: Microorganisms isolated from soil, water, or animal waste.
  • Strain Selection: High-yield producers are selected.
  • Fermentation: Grown in a sterile medium with nutrients.
  • Harvesting: Enzyme extracted via centrifugation/filtration.
• Purification: Using ammonium sulfate precipitation, ion-exchange, or affinity chromatography.
• Stabilization: Adding stabilizers (e.g., glycerol, sucrose) to maintain activity.
• Formulation: Prepared as a liquid or lyophilized powder

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