Purification and partial characterization of a det

Authors: Sib Sankar Giri V Sukumaran Shib Sankar Sen M Oviya B Nazeema Banu Prasant Kumar Jena

Publish Date: 2011/06/30

Volume: 49, Issue: 3, Pages: 455-461

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Abstract

An extracellular detergent tolerant protease producing strain VSG4 was isolated from tropical soil sample and identified as Bacillus subtilis based on morphological biochemical characteristics as well as 16SrRNA gene sequencing The VSG4 protease was purified to homogeneity using ammonium sulphate precipitation dialysis and sephadex G200 gel permeation chromatography with a 174 purification fold The purified enzyme was active and stable over a broad range of pH 80–110 optimum at 90 and temperature 40°C to 60°C optimum at 50°C The thermostability of the enzyme was significantly increased by the addition CaCl2 This enzyme was strongly inhibited by PMSF and DFP suggesting that it belongs to the serine protease superfamily The purified VSG4 alkaline protease showed remarkable stability in anionic 5 mM SDS and ionic 1 Trion X100 and 1 Tween 80 detergents It retained 97±2 and 836±11 of its initial activity after 1 h preincubation in the presence of 1 H2O2 and 1 sodium perborate respectively Furthermore the purified enzyme showed excellent stability and compatibility with some commercial laundry detergents besides its stain removal capacity Considering these promising properties VSG4 protease may find tremendous application in laundry detergent formulations

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