Purification and Characterization of a Thermostable Neutral Metalloprotease I from Chloroflexus aurantiacus J-10-fl
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概要
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Chloroflexus aurantiacus J-10-fl was found to contain two types (protease I and protease II) of thermostable proteases which were separated by Butyl-Toyopearl 650 M chromatography. Protease I was purified to electrophoretic homogeneity from the culture broth of C. aurantiacus J-10-fl. The molecular mass of protease I was estimated to be approximately 66 kDa by SDS-PAGE, and the value of approximately 66 kDa was also obtained by the Hedrick-Smith method, indicating that protease I was a monomer. The isoelectric point was 6.2. Protease I activity was inhibited by metalloprotease inhibitors such as EDTA, EGTA, and o-phenanthroline. The optimum pH for the activity of protease I was around 8.0. Addition of Ca^<2+> increased the pH and heat stabilities of protease I. The activity was stable between pH 4.0-11.0 and up to 75℃, and the maximum activity was observed at 70℃ in the presence of 2 mM CaCl_2. Protease I was resistant to the treatment by denaturing reagents (8 M urea or 1% SDS) at pH 8.0 and 20℃ for 24 h. The sites of cleavage. in oxidized insulin B chain by protease I were similar to those by other microbial neutral metalloproteases. Elastase activity of protease I was not detected.
- 社団法人日本農芸化学会の論文
- 1993-12-23
著者
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Watanabe Akira
Department of Biological Sciences, Graduate School of Science, University of Tokyo
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IZAKI Kazuo
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
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KAMIO Yoshiyuki
Department of Applied Microbiology, Graduate School of Agricultural Science, Tohoku University
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Izaki Kazuo
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Kamio Yoshiyuki
Department Of Agricultural Chemistry Tohoku University
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Kimura Wakako
Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University
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Watanabe Akira
Department Of Biological Sciences Graduate School Of Science The University Of Tokyo
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Kimura Wakako
Department Of Applied Biological Chemistry Faculty Of Agriculture Tohoku University
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Watanabe Akira
Department Of Applied Biological Chemistry Faculty Of Agriculture Tohoku University
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