Characterization of detergent compatible protease of a halophilic Bacillus sp EMB9: Differential role of metal ions in stability and activity

被引：58

作者：

Sinha, Rajeshwari ^{[1
]}

Khare, S. K. ^{[1
]}

机构：

[1] Indian Inst Technol, Dept Chem, Enzyme & Microbial Biochem Lab, New Delhi 110016, India

来源：

BIORESOURCE TECHNOLOGY | 2013年 / 145卷

关键词：

Halophilic; Protease; Solvent stable; Circular dichroism; Salt modulation; SERINE-PROTEASE; PURIFICATION; SOLVENT;

D O I：

10.1016/j.biortech.2012.11.024

中图分类号：

S2 [农业工程];

学科分类号：

0828 ;

摘要：

A moderately halophilic protease producer, Bacillus sp. strain isolated from sea water is described. The protease is purified to homogeneity by ammonium sulphate precipitation and CM cellulose chromatography. The serine protease has a molecular mass of 29 kDa. Enzymatic characterization of protease revealed K-m 2.22 mg mL(-1), V-max 1111.11 U mL(-1), pH optimum 9.0, t(1/2) 190 min at 60 degrees C and salt optima 1% (w/v) NaCl. The protease is remarkably stable in hydrophilic and hydrophobic solvents at high concentrations. The purified preparation is unstable at room temperature. Ca2+ ions are required for preventing this loss of activity. Interestingly, the activity and stability are modulated differentially. Whereas, divalent cation Ca2+ are involved in maintaining stability in solution at room temperature by preventing unfolding, monovalent Na+ and K+ ions participate in regulating the activity and assist in refolding of the enzyme. Application of the protease is shown in efficient removal of blood stain. (C) 2012 Elsevier Ltd. All rights reserved.

引用

页码：357 / 361

页数：5

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