Role of Divalent Metal Ions on Activity and Stability of Thermostable Dipeptidase from Bacillus stearothermophilus
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概要
- 論文の詳細を見る
Thermostable dipeptidase from Bacillus stearothermophilus, a typical metalloenzyme containing 1.0 g atom of Zn per mole of subunit of the dimeric enzyme was markedly activated by exogenous divalent metal ions such as Mn^<2+>, Co^<2+>, and Cd^<2+>. In contrast, several others including Ba^<2+>, Hg^<2+>, and Cu^<2+> considerably, inhibited the enzyme, even the inherent metal, Zn^<2+>, being slightly inhibitory. To study the metal-binding properties of this dipeptidase, the enzyme was completely resolved to the inactive, Zn-free apoenzyme by treatment with EDTA in the presence of guanidine hydrochloride in a weakly acidic buffer. The apoenzyme was readily reconstituted by incubation with either Zn^<2+>, Mn^<2+>, or Co^<2+>, restoring the catalytic activity. The Mn-reconstituted enzyme had nearly twice the activity of the original Zn-enzyme. Combined with kinetic analyses of reconstitution of the apoenzyme with metal ions, these results show that the enzyme has two non-identical metal-binding sites, each with a different property. Furthermore, substitution of Mn^<2+> or Co^<2+> for Zn^<2+> considerably lowered the thermostability of the enzyme without affecting the overall conformation of the enzyme protein, suggesting that the prosthetic Zn is playing dual roles in conformational stability and catalysis of the thermostable dipeptidase.
- 社団法人日本農芸化学会の論文
- 1997-10-23
著者
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SODA Kenji
Department of Biotechnology, Faculty of Engineering, Kansai University
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Soda K
Kansai Univ. Osaka Jpn
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Soda Kenji
Department Of Biotechnology Kansai University
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Soda Kenji
Institute For Chemical Research Kyoto University
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Soda Kenji
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University
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Soda Kenji
Department Of Biotechnology Faculty Of Engineering Kansai University
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Soda Kenji
Institute For Chemical Resarch Kyoto University
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Cho H‐y
Korea Univ. Seoul Kor
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Cho Hong-yon
Department Of Food Sience And Biotechnology Graduate School Of Biotechnology Korea University
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TANIZAWA Katsuyuki
Division of Biological Science, Institute of Scientfic and Industrial Research, Osaka University
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Soda K
Department Of Biotechnology Kansai University
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Cho Hong-yon
Department Of Food Science And Biotechnology Graduate School Of Biotechnology Korea University
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Tanizawa Katsuyuki
Division Of Biological Science Institute Of Scientfic And Industrial Research Osaka University
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Cho Hong-yon
Department Of Food And Biotechnology Korea University
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Soda Kenji
Department of Bioresources Chemistry, Graduate School of Natural Science and Technology, Okayama University
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