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KMID : 0545120150250081281
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Journal of Microbiology and Biotechnology
2015 Volume.25 No. 8 p.1281 ~ p.1290
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Identification and Characterization of a New Alkaline Thermolysin-Like Protease, BtsTLP1, from Bacillus thuringiensis Serovar Sichuansis Strain MC28
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Zhenghong Zhang
Helong Hao
Zhongmei Tang
Zhengzheng Zou
Keya Zhang
Zhiyong Xie
Lilia Babe
Frits Goedegebuur
Xiaogang Gu
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Abstract
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Thermolysin and its homologs are a group of metalloproteases that have been widely used in both therapeutic and biotechnological applications. We here report the identification and characterization of a novel thermolysin-like protease, BtsTLP1, from insect pathogen Bacillus thuringiensis serovar Sichuansis strain MC28. BtsTLP1 is extracellularly produced in Bacillus subtilis, and the active protein was purified via successive chromatographic steps. The mature form of BtsTLP1 has a molecule mass of 35.6 kDa as determined by mass spectrometry analyses. The biochemical characterization indicates that BtsTLP1 has an apparent Km value of 1.57 mg/ml for azocasein and is active between 20oC and 80oC. Unlike other reported neutral gram-positive thermolysin homologs with optimal pH around 7, BtsTLP1 exhibits an alkaline pH optimum around 10. The activity of BtsTLP1 is strongly inhibited by EDTA and a group of specific divalent ions, with Zn2+ and Cu2+ showing particular effects in promoting the enzyme autolysis. Furthermore, our data also indicate that BtsTLP1 has potential in cleaning applications.
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KEYWORD
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Bacillus thuringiensis, Thermolysin-like protease, Alkaline metalloprotease, Thermolysin
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