448 In vitro analysis of the roles of a disulfide bridge and a calcium binding site in Pseudomonas sp. KWI-56 lipase using PCR and Escherichia coli coupled transcription/translation
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概要
- 論文の詳細を見る
The expression of lipase from Pseudomonas sp. KWI-56 had been found to depend on an activator gene. In this work, the active mature lipase was synthesized in vitro using an E. coli S30 coupled transcription/translation system by expressing the lipase gene with its purified activator, or by coexpression of the lipase and the activator gene. The in vitro expression systems were used for studying the folding process of the lipase. The addition of dithiothreitol in the expression systems decreased the lipase activity dramatically without affecting the its synthesis level. And substitutions of Cys190 and Cys270 to serine residues decreased the lipase activity greatly, suggesting that the disulfide bond was essential for the activation of the lipase. In addition, substituting Asp242 and Asp288 also decreased the activities of the in vitro synthesized lipases greatly. The role of the Ca^<2+> binding site in its activation was also discussed.
- 社団法人日本生物工学会の論文
- 1999-08-16
著者
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Yang Junhao
Laboratory Of Molecular Biotechnology Graduate School Of Biological & Agricultural Sciences Nago
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Iwasaki Y
Lab. Of Molecular Biotechnology Dep. Of Bioengineering Sciences Graduate School Of Bioagricultural S
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Iwasaki Yugo
Grad. Sch. Of Bio- Agro-sci. Nagoya Univ.
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Iwasaki Yugo
Laboratory Of Molecular Biotechnology Graduate School Of Bio- And Agro-sciences Nagoya University
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Iwasaki Yugo
Department Of Applied Biological Sciences Faculty Of Agriculture Nagoya University
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Yamane T
Laboratory Of Molecular Biotechnology Graduate School Of Bioagricultural Sciences Nagoya University:
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Yamane Tsuneo
Laboraotory Of Bioreaction Engineering Department Of Food Science And Technology Faculty Of Agricult
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Yamane Tsuneo
Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur
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Kobayashi K
Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University
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Kohda K
Toyota Central R&d Laboratories Inc.
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Yamane T
Laboratory Of Molecular Biotechnology Graduate School Of Biological & Agricultural Sciences Nago
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Yamane T
Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur
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Kobayashi K
Laboratory Of Molecular Biotechnology Graduate School Of Biological & Agricultural Sciences Nago
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NAKANO Hideo
Grad. Sch. Bioagri. Sci., Nagoya Univ.
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Kobayashi K
Shizuoka Univ. Shizuoka Jpn
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Yang Junhao
Grad. Sch. Bio. Agro-Sci., Nagoya Uni.
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Kobayashi Koei
Grad. Sch. Bio. Agro-Sci., Nagoya Uni.
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Yamane Tsuneo
Grad. Sch. Bio. Agro-Sci., Nagoya Uni.
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Yabuki Takashi
Protein Research Group Riken Genomic Sciences Center
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Yamane Tsuneo
Dept. Of Food Science And Technology Faculty Of Agriculture Nagoya University
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Yamane Tsuneo
Laboratory Of Molecular Biotechnology Graduate School Of Bio- And Agro-sciences Nagoya University
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Iwasaki Yugo
Grad. Sch. Bioagric., Sci., Nagoya Univ.
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