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Biochemical Characterization of a Thiol-Activated, Oxidation Stable Keratinase from Bacillus pumilus KS12.

ABSTRACT: An extracellular keratinase from Bacillus pumilus KS12 was purified by DEAE ion exchange chromatography. It was a 45?kDa monomer as determined by SDS PAGE analysis. It was found to be an alkaline, serine protease with pH and temperature optima of 10 and 60°C, respectively. It was thiol activated with two- and eight-fold enhancement in presence of 10 mM DTT and ?-mercaptoethanol, respectively. In addition, its activity was stimulated in the presence of various surfactants, detergents, and oxidizing agents where a nearly 2- to 3-fold enhancement was observed in presence of H(2)O(2) and NaHClO(3). It hydrolyzed broad range of complex substrates including feather keratin, haemoglobin, fibrin, casein,and ?-keratin. Analysis of amidolytic activity revealed that it efficiently cleaved phenylalanine ? leucine ? alanine- p-nitroanilides. It also cleaved insulin B chain between Val(2)- Asn(3), Leu(6)-Cys(7) and His(10)-Leu(11) residues.

SUBMITTER: Rajput R

PROVIDER: S-EPMC2956970 | biostudies-literature | 2010 Jul

REPOSITORIES: biostudies-literature

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Biochemical Characterization of a Thiol-Activated, Oxidation Stable Keratinase from Bacillus pumilus KS12.

Rajput Rinky R Sharma Richa R Gupta Rani R

Enzyme research 20100715

An extracellular keratinase from Bacillus pumilus KS12 was purified by DEAE ion exchange chromatography. It was a 45 kDa monomer as determined by SDS PAGE analysis. It was found to be an alkaline, serine protease with pH and temperature optima of 10 and 60°C, respectively. It was thiol activated with two- and eight-fold enhancement in presence of 10 mM DTT and β-mercaptoethanol, respectively. In addition, its activity was stimulated in the presence of various surfactants, detergents, and oxidizi ...[more]

PMID: 21048858

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