Overproduction of DnaJ in Escherichia coli Improves in Vivo Solubility of the Recombinant Fish-derived Transglutaminase
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概要
- 論文の詳細を見る
The overexpression of red sea bream (Pagrus major) transglutaminase (TGase, E. C. 2. 3. 2. 13) in Escherichia coli mostly leads to the accumulation of biologically inactive enzyme. Although the solubility of the gene products could be improved by cultivation at a lower temperature (26-28℃), most of the synthesized TGase was still in the form of insoluble aggregates. The effects of overproduction of molecular chaperones on the intracellular solubility of newly produced recombinant TGase were examined. The overexpression of dnaK or groES/EL did not improve solubility. However, DnaJ greatly increased the solubility of the recombinant TGase, resulting in active enzyme in the presence of calcium ions. Co-expression of dnaK along with dnaJ further increased the content of soluble TGase. Under our experimental conditions, supplementation with both DnaJ and DnaK elevated the TGase activity in the producer cells by roughly 4-fold, compared with the control strain cultured at 30℃. Thus, we found that DnaJ is important in controlling the solubility of protein overproduced in E. coli.
- 社団法人日本農芸化学会の論文
- 1998-06-23
著者
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Yoon Ki
Department Of Biotechnology The University Of Tokyo
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Kikuchi Yoshimi
Central Research Laboratories Ajinomoto Co. Inc.
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Yokoyama Keiichi
Department Of Pathology And Cell Regulation Kyoto Prefectural University Of Medicine
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Yokoyama Keiichi
Department Of Biotechnology Graduate School Of Agricultre And Life Science The University Of Tokyo
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Yokoyama Keiichi
Food Research & Development Laboratories
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YOKOYAMA Keiichi
Institute of Life Sciences, Ajinomoto Co., Inc.
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Yokoyama K
Central Research Laboratories Ajinomoto Co. Inc.
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YASUEDA Hisashi
Central Research Laboratories, Ajinomoto Co. Inc.,
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Yokoyama Keiichi
京都府立医科大学 第2病理
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Yasueda Hisashi
Central Research Laboratories Ajinomoto Co. Inc.
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Yokoyama Keiichi
Institute Of Life Sciences Ajinomoto Co. Inc.
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Yokoyama Kei-ichi
Food Research & Development Laboratories
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