Over-production of Thermus protease in dense culture of Escherichia coli using two carbon sources |
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| Authors: | Kazumasa Tsuji Masaharu Iijima Hiroshi Matsuzawa Shinji Sakamoto |
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| Institution: | (1) Sea Water Science Research Laboratory, Japan Tobacco Inc., 4-13-20 Sakawa, Odawara Kanagawa, 256, Japan;(2) Life Science Research Laboratory, Japan Tobacco Inc., 6-2 Umegaoka, Aoba-ku Yokohama, 227, Japan;(3) Department of Agricultural Chemistry, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku Tokyo, 113, Japan;(4) Present address: Pharmaceutical Frontier Research Laboratories, Japan Tobacco Inc., 6-2 Umegaoka, Aoba-ku Yokohama, 227, Japan |
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| Abstract: | Escherichia coli JM109(DE3) harboring expression plasmid pkAQNÆC30, which carries the Thermus protease aqualysin I (AQI) gene, was cultivated with glucose as a sole carbon source. The final cell concentration was over 15 g dry weight/l and the amount of AQI produced reached approximately 130 kU/ml broth. Moreover, by using two carbon sources, glucose and glycerol, the production yield was increased to over 200 kU AQI/ml, while suppressing the formation of inhibitory acetic acid. |
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