AlkalineProteas

e(Fermented

Leather Enzyme)

Submitted By, Effat Jahan Tamanna

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