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Purification and characterization of organic solvent stable serine alkaline protease from newly isolated Bacillus circulans M34
Author(s) -
Sari Esma,
Loğoğlu Elif,
Öktemer Atilla
Publication year - 2015
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3431
Subject(s) - bacillus circulans , chemistry , protease , chromatography , serine protease , affinity chromatography , sepharose , enzyme , biochemistry
A protease from newly isolated Bacillus circulans M34 was purified by Q‐Sepharose anion exchange chromatography and Sepharose–bacitracin affinity chromatography followed by (NH 4 ) 2 SO 4 precipitation. The molecular mass of the purified enzyme was determined using SDS–PAGE. The optimum pH and temperature for protease activity were 11 and 50°C, respectively. The effect of various metal ions on protease activity was investigated. Alkaline protease from Bacillus circulans M34 wase activated by Zn 2+ , Cu 2+ and Co 2+ up to 31%. The purified protease was found to be stable in the organic solvents, surfactants and oxidizing agent. The substrate specificity of purified protease was investigated towards different substrates. The protease was almost completely inhibited by the serine protease inhibitor phenylmethanesulfonyl fluoride. The kinetic parameters of the purified protease, maximum rate ( V max ) and Michaelis constant ( K m ), were determined using a Lineweaver–Burk plot. Copyright © 2015 John Wiley & Sons, Ltd.