Dependence of Hydrolysis of β-Lactams with a Zinc(II)-β-Lactamase Produced from Sermtia marcescens (IMP-1) on pH and Concentration of Zinc(II) Ion : Dissociation of Zn(II) from IMP-1 in Acidic Medium
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概要
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The pH dependence for the hydrolysis of β-lactam antibiotics by a metallo-β-lactamase (IMP-1) produced from Serratia marcescens was investigated varying the concentration of Zn(II). The activity of IMP-1 for imipenem was decreased at pH less than pH 5.3 without external addition of Zn(II) ions but was recovered with addition of Zn(II), Varying the concentration of external Zn(II), the molar activity of the enzyme, k_<obs>, that was defined by the velocity of hydrolysis of imipenem/concentration of IMP-1 was expressed by k_<obs>=v_<init>/[E]_T=k_<max>[Zn]/(k_d+[Zn]) in which X_d stands for the dissociation constant between Zn(II) and IMP-1. The dissociation constants, K_d, vary with pH; A_d = 840×10^<-6>_M at pH 4.3 and k_d = 0.19×10^<-6>_M at pH 6,0. The plot of -logk_d against pH showed a straight line having a slope of 4.0 below pH 5.0, showing the existence of four functional groups which may be protonated upon dissociation of Zn(II) ion(s). The k_<cat>, K_m, and k_<eat>/K_m of hydrolysis of imipenem and cephalothin in the presence of sufficient concentration of Zn(NO_3)_2 for saturation of IMP-1 with Zn(II) showed similar dependency to each other on pH between pH 6.0 and 9.0.
- 公益社団法人日本薬学会の論文
- 2003-05-01
著者
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Goto M
School Of Pharmaceutical Sciences Kitasato University
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Miura Hirohito
National Food Research Institute
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Yamaguchi Y
The Kitasato Institute
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Higashi T
Toyama Medical And Pharmaceutical Univ. Toyama Jpn
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Higashi T
Faculty Of Pharmaceutical Sciences Kanazawa University
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Nakayama H
Laboratory Of Biofunctional Chemistry Faculty Of Pharmaceutical Sciences Kumamoto University
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Goto M
Graduate School Of Pharmaceutical Sciences Kumamoto University
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ARAKAWA Yoshichika
Department of Bacterial and Blood Products, National Institute of Health
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Moriya H
Department Of Pathology Gifu University School Of Medicine
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Yamaguchi Y
Environmental Safety Center Kumamoto University
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YASUZAWA Hisami
Graduate School of Pharmaceutical Sciences, Kumamoto University
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GOTO Masafumi
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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YASUZAWA Hisami
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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HIGASHI Toshihiro
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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YAMAGUCHI Yoshihiro
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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KAWANAMI Akiko
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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MIFUNE Shiho
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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MORI Hiromasa
Laboratory of Pharmaceutical Physical Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto Univer
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NAKAYAMA Hitoshi
Laboratory of Biofunctional Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto University
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HARADA Kumiko
Laboratory of Biofunctional Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto University
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森 春樹
Research Center Mitsui Toatsu Chemicals
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Mifune Shiho
Laboratory Of Pharmaceutical Physical Chemistry Faculty Of Pharmaceutical Sciences Kumamoto Universi
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Harada Kumiko
Laboratory Of Biofunctional Chemistry Faculty Of Pharmaceutical Sciences Kumamoto University
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Kawanami Akiko
Laboratory Of Pharmaceutical Physical Chemistry Faculty Of Pharmaceutical Sciences Kumamoto Universi
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Yasuzawa Hisami
Graduate School Of Pharmaceutical Sciences Kumamoto University
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Arakawa Y
Department Of Bacterial Pathogenesis And Infection Control National Institute Of Infectious Diseases
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Arakawa Yoshichika
Department Of Bacterial And Blood Products National Institute Of Health
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Yamaguchi Yoshihiro
Environmental Safety Center Kumamoto University
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Yamaguchi Yoshihiro
Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University
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