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Alkaline Protease• One of the class of protease enzyme.• An extracellular enzyme.• Performs proteolysis, that is, protein catabolism by hydrolysis of
the peptide bonds. • Active at alkaline pH 8 to 12 and at temperature 30⁰-80⁰C.• Molecular weight is about 20,000 to 45,000 Dalton.• The structure is determined by X-ray crystallography. • EC (Enzyme Commission) Number: 3.4.21–24.99• In 1971, Japanese scientist Koki Horikoshi first reported the
production of alkaline protease from bacteria.
Sources of Alkaline ProteaseBacteria Fungi
Bacillus subtilis Aspergillus flavus
Bacillus pumilus Aspergillus fumigatus
Bacillus licheniformis
Aspergillus melleus
Bacillus altitudinis Aspergillus niger
Bacillus firmus Fusarium graminearum
Bacillus amyloliquefaciens
Penicillium griseofulvim
Bacillus proteolyticus
Penicillium lilacinus
Thermomonospora fusca
Scedosporium apiosermum
Genetic Engineering of Microbes• More than 50% of the industrially important
enzymes are now produced from genetically engineered microorganisms.
• Methods Used: Conventional mutagenesis (UV or chemical exposure) or Recombinant DNA technology.
Production Process of Alkaline Protease Enzyme• Isolation of Microorganisms• Development and Preparation of Inoculum• Preparation of Fermentation Media• Optimization of Media• Fermentation Process• Enzyme Extraction and Assay• Protein Assay• Alkaline Protease Purification
Ammonium sulphate precipitation Ultracentrifugation Flocculation Chromatography Electrophoresis
• Characterization of Purified Alkaline Protease• Packaging
Alkaline Protease in Leather IndustryStages Enzymes involved Function of
EnzymesCuring Enzymes are not directly
involvedTo preserve hides and skins
Soaking Alkaline Protease To remove non fibrillar proteins
Dehairing Neutral and Alkaline Protease
To remove hair
Bating Trypsin and Alkaline Protease
To make soft, supple and pliable
Degreasing
Lipase and Acid Protease To remove fats
Tanning Enzymes are not directly involved
To influence the quality of tanning
Waste Processin
g
Trypsin and Alkaline Protease
Tanned waste processing
Advantages of Using Enzymes in Leather Industry
• Significant reduction of using chemicals.• Simplification of processes.• Creates of an ecologically conducive atmosphere for the workers.• Leathers have shown better strength and quality.• Saves time.• Environment friendly.• Leather wastes can be hydrolyzed by enzymes.
Global Market of Alkaline Protease Enzyme• Of the industrial enzymes, Alkaline protease covers 25% of
total enzyme market.• The global market is increasing because
replacement of chemicals and additives, more demand for environment friendly ingredients, less production cost, high yield, less time taken for production and the growing industrial need in developing countries.
Companies Mentioned:• AB Enzymes, Germany • Novozymes, Denmark• Amano Enzymes, Japan• Dyadic International, Inc. USA • Jiangsu Boli Bioproducts Co. Ltd, China• Advanced Enzymes, India