Expression of Aqualysin I (a Thermophilic Protease) in Soluble Form in Escherichia coli under a Bacteriophage T7 Promoter
スポンサーリンク
概要
- 論文の詳細を見る
The thermophilic protease aqualysin I (AQI) gene (aqul), derived from Thermus aquaticus YT-1, was inserted under the control of the bacteriophage T7 promoter in an expression plasmid. The plasmid was introduced into two strains of E. coli JM109 (DE3), one carrying and one lacking an F' episome, which carries the lacl^q gene. Upon cultivation the strain carrying an F' episome produced AQI as an insoluble fusion protein (74kDa) with the T7 gene 10 protein. This insoluble protein could not be processed into mature AQI by heat treatment and thus it had no proteolytic activity. On the other hand, when the strain lacking an F' episome was used as a host cell for aqul expression, non-induced, or leaky, expression occurred, and AQI was produced in a soluble form. This soluble protein could be processed into active AQI by heat treatment. Moreover, when a low concentration of IPTG (0.0125mM) was added, the amount of active AQI was 2.7 times greater than that produced in a batch culture without induction.
- 社団法人日本農芸化学会の論文
- 1995-08-23
著者
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Ohta T
Hokkaido Food Processing Res. Center Hokkaido Jpn
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Mori Hiroaki
Department Of Agricultural Chemistry The University Of Tokyo
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MATSUZAWA Hiroshi
Department of Biotechnology, The University of Tokyo
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Sakamoto Shinji
Life Science Research Laboratory Japan Tobacco Inc.
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OHTA Takahisa
Department of Agricultural Chemistry, The University of Tokyo
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Ohta Takahisa
Department Of Agricultural Chemistry The University Of Tokyo
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TERADA Ichiro
Sea Water Science Research Laboratory, Japan Tobacco Inc.
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Terada Ichiro
Sea Water Science Research Laboratory Japan Tobacco Inc
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Matsuzawa H
Department Of Bioscience And Biotechnology Aomori University
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Iijima Masaharu
Life Science Research Laboratory, Japan Tobacco Inc.
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Iijima M
Life Science Research Laboratory Japan Tobacco Inc.
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Matsuzawa Hiroshi
Department Of Biotechnology And Biotechnology Research Center The University Of Tokyo
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Matsuzawa Hiroshi
Department of Bioscience and Biotechnology, Aomori University
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MATSUZAWA Hiroshi
Department of Agricultural Chemistry, The University of Tokyo
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