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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Acknowledgments
The authors thank Professor E. Fukusaki (Osaka University) for his kind help with peptide mass fingerprinting using LC-MS/MS. This work was supported by a Grant-in-Aid for Scientific Research (B) to H.S. from the Japan Society for the Promotion of Science (no. 21360401) and a Grant-in-Aid for Young Scientists (B) to T.H. from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (no. 21780071). This work was also supported in part by the Global COE Program from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.
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J. Kim and H. Fukuda equally contributed to this work.
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Kim, J., Fukuda, H., Hirasawa, T. et al. Requirement of de novo synthesis of the OdhI protein in penicillin-induced glutamate production by Corynebacterium glutamicum . Appl Microbiol Biotechnol 86, 911–920 (2010). https://doi.org/10.1007/s00253-009-2360-6
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DOI: https://doi.org/10.1007/s00253-009-2360-6