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- Title
Characterization of a Novel Organic Solvent Tolerant Protease from a Moderately Halophilic Bacterium and Its Behavior in Ionic Liquids.
- Authors
Karbalaei-Heidari, Hamid; Shahbazi, Mahnaz; Absalan, Ghodratollah
- Abstract
An extracellular protease was purified from a novel moderately halophilic bacterium Salinivibrio sp. strain MS-7 by the combination of an acetone precipitation (40-80 %) step and a DEAE-cellulose anion exchange column chromatography. Kinetic parameters of the enzyme exhibited V and K of 130 U/mg and 1.14 mg/ml, respectively, using casein as a substrate. The biochemical properties of the enzyme revealed that the 21-kDa protease had a temperature and pH optimum of 50 °C and 8.0, respectively. The enzyme was strongly inhibited by phenylmethylsulfonyl fluoride, Pefabloc SC, chymostatin, and also EDTA, indicating that it belongs to the class of serine metalloproteases. Interestingly, Ba and Ca (2 mM) strongly enhanced the enzyme activity, while Fe and Mg activated moderately and Zn, Ni, and Hg decreased the enzyme activity. The effect of organic solvents with different log P on the purified protease revealed complete stability in toluene, ethyl acetate, chloroform, and n-hexane at 10 and 50 % ( v/ v) and moderate stability even in 50 % of DMSO and ethanol. The behavior of the MS-7 protease in three imidazolium-based ionic liquids exhibited suitable activity in these green solvent systems, especially in 1-hexyl-3-methylimidazolium hexafluorophosphate ([CMIM][PF]). Comparison of the purified protease with other previously reported proteases suggests that strain MS-7 secrets a novel organic solvent-tolerant protease with outstanding activity in organic solvents and imidazolium-based ionic liquids, which could be applied in low water synthetic section of industrial biotechnology.
- Subjects
ORGANIC solvents; PROTEOLYTIC enzymes; HALOBACTERIUM; IONIC liquids; ENZYMES
- Publication
Applied Biochemistry & Biotechnology, 2013, Vol 170, Issue 3, p573
- ISSN
0273-2289
- Publication type
Academic Journal
- DOI
10.1007/s12010-013-0215-1