Purification and characterization of a thermo- and organic solvent-tolerant alkaline protease from Bacillus sp. JER02.
Bacillus sp. JER02 is a bacterial strain that can be grown in a medium containing organic solvents and produce a protease enzyme. JER02 protease was purified with a yield of 31.9% of total protein and 328.83-fold purification. Km and Vmax of this protease were established as 0.826 µM and 7.18 µmol/min, respectively. JER02 protease stability was stimulated about 80% by cyclohexane. It exhibited optimum temperature activity at 70°C. Furthermore, this enzyme was active in a wide range of pH (4-12) and showed maximum activity at pH 9.0. The nonionic detergents Tween-20 and Triton X-100 improved the protease activity by 30 and 20%, respectively. In addition, this enzyme was shown to be very stable in the presence of strong anionic surfactants and oxidizing agents, since it retained 77%, 93%, and 98% of its initial activity, after 1 hr of incubation at room temperature with sodium dodecyl sulfate (SDS), sodium perborate (1%, v/v) and H2O2 (1%, v/v), respectively. Overall, the unique properties of the Bacillus sp. JER02 protease suggested that this thermo- and detergent-stable, solvent-tolerant protease has great potential for industrial applications.
a Department of Biology, Faculty of Sciences , Shahid Bahonar University of Kerman , Kerman , Iran.
Arastoo
A
Badoei-Dalfard
Zahra
Z
Karami
Hadi
H
Ravan
D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003902:Detergents / Q000737:chemistry; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D004795:Enzyme Stability; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000737:chemistry; D012997:Solvents / Q000737:chemistry*
D001426:Bacterial Proteins; D003902:Detergents; D012997:Solvents; D006861:Hydrogen Peroxide; D010450:Endopeptidases; C087822:alkaline protease
characterization; organic solvent; protease; purification; thermo-stable