Requirement of de novo synthesis of the OdhI protein in penicillin-induced glutamate production by Corynebacterium glutamicum |
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Authors: | Jongpill Kim Hirohisa Fukuda Takashi Hirasawa Keisuke Nagahisa Kazuo Nagai Masaaki Wachi Hiroshi Shimizu |
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Institution: | (1) Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita Osaka, 565-0871, Japan;(2) Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita Osaka, 565-0871, Japan;(3) Department of Biological Chemistry, Chubu University, 1200 Matsumoto-cho, Kasugai Aichi, 487-8501, Japan;(4) Department of Bioengineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503, Japan; |
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Abstract: | We found that penicillin-induced glutamate production by Corynebacterium glutamicum is inhibited when a de novo protein synthesis inhibitor, chloramphenicol, is added simultaneously with penicillin. When chloramphenicol
was added 4 h after penicillin addition, glutamate production was essentially unaffected. 3H-Leucine incorporation experiments revealed that protein synthesis continued for 1 h after penicillin addition and then gradually
decreased. These results suggest that de novo protein synthesis within 4 h of penicillin treatment is required for the induction
of glutamate production. To identify the protein(s) necessary for penicillin-induced glutamate production, proteome analysis
of penicillin-treated C. glutamicum cells was performed with two-dimensional gel electrophoresis. Of more than 500 proteins detected, the amount of 13 proteins,
including OdhI (an inhibitory protein for 2-oxoglutarate dehydrogenase complex), significantly increased upon penicillin treatment.
Artificial overexpression of the odhI gene resulted in the decreased specific activity of the 2-oxoglutarate dehydrogenase complex and increased glutamate production
without any triggers. These results suggest that the de novo synthesis of OdhI is the necessary factor for penicillin-induced
glutamate overproduction by C. glutamicum. Moreover, continuous glutamate production was achieved by overexpression of odhI without any triggers. Thus, the odhI-overexpressing strain of C. glutamicum can be useful for efficient glutamate production. |
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