Abstract

ISOLATION, OPTIMIZATION AND CHARACTERIZATION OF ALKALINE PROTEASE PRODUCING BACTERIA FROM COLLAGEN LAYER OF DECAYING SKIN (SHEEP UPPER ) LEATHER INDUSTRY

S.Manikandan, Dr.P.Kavitha, Dr.A.Panneerselvam

Abstract

Enzymes are organic catalysts that play a vital role in many aspects of life. Enzymes play a huge role in day today life from simple fermentation to complicated gene expressions. Protease is enzyme that has the ability to degrade protein by the breaking of the hydrogen bond that bunds protein into peptides and proteins. Microbial proteases belong to acid, neutral or alkaline based on their pH optimum for activity and the active site viz., Metallo (EC 3.4.24), Aspartic (EC 3.4.23) Cysteine or Sulphydryl (EC.3.4.22) or Serine type (EC 3.4.21). Alkaline protease is stable in alkaline pH and possesses a serine residue at the active site. Proteases constitute a class of industrial enzymes which alone form approximately 60% of the total world-wide enzyme production . Among the various proteases microbial proteases play an important role in biotechnological process. Alkaline proteases produced are of special interest as they could be used in the manufacture of detergents, food, pharmaceuticals and leather. Alkaline protease is produced by a wide variety of microbial species like Bacillus subtilis, Aspergillus oryzae, Streptomyces cellulasae and Aeromonas hydrophila species. Most commercial proteases, mainly neutral and alkaline are produced by organisms belonging to the Bacillus sp. Bacillus sp are attractive industrial tools for a variety of reasons, including their high growth rates leading to short fermentation cycle times, their capacity to secrete proteins into the extracellular media and the GRAS (Generally Regarded as Safe) status with food and drug administration for species such as Bacillus subtilis and Bacillus licheniformis . Bacillus sp is used extensively for the production of industrial enzymes such as amylase, lipase and alkaline protease. It is a popular host for industrial preparation of gene products. In our study twenty one (21 No’s) bacterial strains were isolated from the sheep upper collagen layer of decaying skin sample. The fifteen (15 No’s) isolates exhibited proteolytic activity based on clear zone formation on skim milk agar medium at pH 9.0. These isolates were then characterized and identified. Biochemical test revealed the organisms to be Bacillus sp and Pseudomonas sp. Maximum yield of enzyme (22 mg/ml) was obtained in Naser Tanning company Chennai.

Keywords: Alkaline Protease, Sheep Collage Layer, Bacillus sp.