Purification and characterization of organic solvent stable protease from Bacillus licheniformis RSP-09-37

Applied Microbiology and Biotechnology
Ritu Sareen, Prashant Mishra

Abstract

A protease was purified from the cell-free supernatant of Bacillus licheniformis RSP-09-37, a mutant from a thermophilic bacterial strain, B. licheniformis RSP-09, using affinity chromatography with alpha-casein agarose resin. The protease was purified 85-fold to electrophoretic homogeneity. The apparent molecular mass of purified protease was 55 kDa using gel filtration in high-performance liquid chromatography, which is in agreement with the results obtained from sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting a monomeric nature of the protein. The purified protease revealed temperature optima of 50 degrees C and pH optima of 10.0 and was classified as serine protease based on its complete inhibition with phenyl methyl sulfonyl fluoride. The purified protease exhibited tolerance to both detergents and organic solvent. The synthetic activity of the protease was tested using the transesterification reaction between N-acetyl-L-phenylalanine-ethyl ester and n-propanol in organic solvents varying in their log P values and the kinetic parameters of the enzyme in these organic solvents were studied. The enzyme has potential to be employed for synthetic reactions and in detergent formulations.

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Citations

Oct 15, 2013·Applied Biochemistry and Biotechnology·J Deepa Arul PriyaPennathur Gautam
Apr 4, 2012·Annual Review of Chemical and Biomolecular Engineering·Michael J LiszkaDouglas S Clark
Nov 16, 2013·BMC Biochemistry·Prashant T SanatanVandana K Hivrale
Apr 21, 2011·Biotechnology Advances·Sebastian TorresGuillermo R Castro
Mar 31, 2015·Applied Biochemistry and Biotechnology·Aungkawipa MontriwongJirawat Yongsawatdigul
May 23, 2014·Preparative Biochemistry & Biotechnology·Arastoo Badoei-DalfardHadi Ravan
Aug 15, 2017·Journal of Basic Microbiology·Tharcilla B A PessoaJoão C T Dias
Sep 17, 2008·Reproductive Health Matters·Bela Ganatra
Dec 6, 2017·Biotechnology & Genetic Engineering Reviews·Goutam Banerjee, Arun Kumar Ray
Jul 8, 2020·Preparative Biochemistry & Biotechnology·Xincai WuYaowei Fang
Sep 10, 2017·Archives of Microbiology·Abhijit Poddar, Subrata K Das

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