Overexpression and In Vitro Reconstitution of the Ethylene-Forming Enzyme from Pseudomonas syringae
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概要
- 論文の詳細を見る
By replacing a native promotor which lac and tac promoters, the gene encoding an ethylene-forming enzyme (EFE) from Pseudomonas syringae pv. phaseolicola PK2 was overexpressed in Escherichia coli. The EFE protein expressed by a multicopy accounted for more than 30% of the total cellular protein, resulting in ethylene-fortming activities higher than 10 μl of ethylene (mg cell)^<-1>h ^<-1> in recombinant E. coli cells. However, most of the EFE protein accumulated as inactive inclusion bodies particularly at elevated temperatures (>30℃). We present an efficient procedure for reconstituting an active enzyme from inclusion bodies by solubilization with 8 M urea and dialysis. The reconstituted EFE has specific activity identical to that of the native enzyme from P. syringae, suggesting that the EFE protein has an intrinsic folding capability in vitro.
- 社団法人日本生物工学会の論文
- 1995-03-25
著者
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MATSUOKA MASAYOSHI
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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OGAWA TAKAHIRA
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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Inoue Yorinao
The Institute of Physical and Chcmical Research
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Fukuda H
Kobe Univ. Kobe
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Fukuda Hideki
Department Of Molecular Science And Material Engineering Graduate School Of Science And Technology K
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Ogawa T
Kyoto Univ. Kyoto Jpn
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Ogawa Takahiro
Department Of Biological Science And Technology Science University Of Tokyo
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Ogawa T
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Matsuoka M
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Matsuoka Masayoshi
Dept. Applied Microb. Technol. Sojo Univ.
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FUKUDA HIDEO
Department of Applied Microbial Technology, Kumamoto Institute of Technology
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TANASE SUMIO
Department of Biochemistry, Kumamoto University School of Medicine
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ISHIHARA KATSUYA
Department of Applied Microbial Technology, Kumamoto Institute of Technology
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Fukuda H
Kobe Univ. Kobe Jpn
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Ogawa T
Kumamoto Inst. Technol. Kumamoto Jpn
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Ogawa Takahira
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Ogawa Takahira
Dept. Appl. Microbial Technology Kumamoto Institute Of Technology
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Matsuoka Masayoshi
Dept. Appl. Microbial Technology Kumamoto Institute Of Technology
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Matsuoka Masayoshi
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Fukuda Hideo
Dept. Appl. Microbial Technology Kumamoto Institute Of Technology
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Takegawa Sumio
Department Of Microbiology Kinki University School Of Medicine
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Tanase Sumio
Department Of Biochemistry Kumamoto University School Of Medicine
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Ishihara Katsuya
Division Of Cardiology And Cardiac Surgery Kofu Johnan Hospital
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Fukuda Hideo
Department Of Applied Microbial Technology Kumamoto Institute Of Technology
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Inoue Yorinao
The Institute Of Physical And Chemical Research (riken)
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Inoue Yorinao
The Inst. Of Phys. And Chem. Res. (riken)
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