Microbial Protease

Industrial production
Submitted to:
Dr. Pooja Suneja Submitted by:
Associate Professor Yashika rani (2293)
Department of Microbiology PhD coursework
 Enzymes
• Catalyse biochemical reactions
• Lowers activation energy
• Produced by plants/animals/microbes
 Industrial enzymes
• Facilitate industrial process.
• Product production
• Microbial enzymes
• Purified enzyme
• Hydrolases (protease, amylase, lipase)
Technical enzymes Food enzymes
Animal feed enzymes
 Microbial Enzymes
• Economical
• Easily produced and purified
• Variety
• Tolerance to wide range of environmental condition
• Easy genetic manipulation
• Short generation time
 Microbial Protease
• Hydrolysis of peptide bond
• Endopeptidase & Exopeptidase (internal or terminal cleavage)
Protein Endopeptidase Polypeptide
Polypeptide Exopeptidas Amino acids
e
• Second most important enzyme after amylase
• Produced from bacteria or fungus
By Proteolytic mechanism
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Glutamic Proteolytic Cysteine

protease mechanis protease
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 Optimum pH

Neutral protease
Acidic protease Alkaline protease
opH 7
o pH 2-5 o pH 8-11
oBacterial / Fungal
o Fungal origin o Bacterial origin
origin
 Acidic protease
• Animal cell/yeast/moulds
• Rarely in bacteria
• Renin-like enzymes are derived from Mucor michei, Mucor pusillus,
Mucor racemaeus, Mucor bacilliformis. (used in cheese production)
• Pepsin like acid proteases are derived from Aspergillus species and
Rhizopus species.
 Alkaline protease
• Most important producer
Bacillus strain - Bacillus licheniformis, Bacillus amylotiquefaciens, B. megaterium, B.
purilis
Streptomyces strain - Streptomyces fradiae, S. griseus, S. rectus and fungi
like Aspergillus niger, A. sojae, A. oryzae, A. flavus
• Mostly used in detergents
As they are stable at alkaline pH, stable at high temperature, stable association with chelating agents.
 Neutral protease
• Narrow pH range
• Sensitive to increased temperature
• Restricted industrial applications but can be used in leather industries and in food
industries
• Inactivated by alkaline protease
• Neutral proteases are produced by bacteria and fungi. E.g. Bacillus subtilis, B.
coreus, B. megaterium, Pseudomonas aeruginosa, Streptomyces griseus,
Aspergillus oryzae, Aspergillus sojae.
 Production

• Isolation
Bacillus species are specific producer of extracellular proteases. Proteolytic
microbes can be isolated by observing hydrolysis in casein agar.
• Media formation
Media rich in nitrogen sources such as soyabean milk, casein, gelatine and
carbohydrate sources such as starch, or lactose are generally used for
protease production.
• Fermentation
Inoculation , Incubation, Production phase, Harvesting.
Initial growth is carried out in shaken flask and small fermenter at 30-37 oC
• Purification
Precipitation
Ultrafiltration
Chromatographic technique (ion exchange) purification by treatment with activated charcoal
and H2O2
 Application
• Dairy Industry
• Beer Industry
• Wine Industry
• Laundry and dishwashing detergent
• Food processing
• Bioremediation
• Textile industries
 References
• Hailemichael, F. Production and Industrial Application of Microbial
Aspartic Protease: A Review.
• Zhu, D., Wu, Q., & Wang, N. (2011). 3.02 – Industrial Enzymes.
THANK YOU