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http://dx.doi.org/10.4014/jmb.1009.09001

Purification and Characterization of Manganese-Dependent Alkaline Serine Protease from Bacillus pumilus TMS55  

Ibrahim, Kalibulla Syed (Department of Biotechnology, Alagappa University)
Muniyandi, Jeyaraj (Department of Biotechnology, Alagappa University)
Pandian, Shunmugiah Karutha (Department of Biotechnology, Alagappa University)
Publication Information
Journal of Microbiology and Biotechnology / v.21, no.1, 2011 , pp. 20-27 More about this Journal
Abstract
The purification and characterization of a $Mn^{2+}$-dependent alkaline serine protease produced by Bacillus pumilus TMS55 were investigated. The enzyme was purified in three steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by gel filtration and cation-exchange chromatography. The purified protease had a molecular mass of approximately 35 kDa, was highly active over a broad pH range of 7.0 to 12.0, and remained stable over a pH range of 7.5 to 11.5. The optimum temperature for the enzyme activity was found to be $60^{\circ}C$. PMSF and AEBSF (1 mM) significantly inhibited the protease activity, indicating that the protease is a serine protease. $Mn^{2+}$ ions enhanced the activity and stability of the enzyme. In addition, the purified protease remained stable with oxidants ($H_2O_2$, 2%) and organic solvents (25%), such as benzene, hexane, and toluene. Therefore, these characteristics of the protease and its dehairing ability indicate its potential for a wide range of commercial applications.
Keywords
Bacillus pumilus; alkaline serine protease; dehairing; manganese dependent; enzyme purification;
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