Requirement of de novo synthesis of the OdhI protein in penicillin-induced glutamate production by Corynebacterium glutamicum
Requirement of de novo synthesis of the OdhI protein in penicillin-induced glutamate production by Corynebacterium glutamicum
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DOI：
10.1007/s00253-009-2360-6
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发表时间：
2010-04-01
影响因子：
5
通讯作者：
Shimizu, Hiroshi
中科院分区：
文献类型：
--
作者：
Kim, Jongpill;Fukuda, Hirohisa;Shimizu, Hiroshi
我们发现，青霉素诱导谷氨酸棒杆菌谷氨酸生产抑制时，从头蛋白质合成抑制剂，氯霉素，同时加入青霉素。当氯霉素加入青霉素后4小时，谷氨酸生产基本上不受影响。H-3-亮氨酸掺入实验表明，蛋白质的合成持续1小时后，青霉素添加，然后逐渐减少。这些结果表明，从头蛋白质合成4小时内青霉素治疗所需的诱导谷氨酸生产。为了鉴定青霉素诱导谷氨酸产生所必需的蛋白质，对青霉素处理的C。用双向凝胶电泳进行谷氨酸细胞。在检测到的500多种蛋白质中，包括OdhI（2-酮戊二酸脱氢酶复合物的抑制蛋白）在内的13种蛋白质的量在青霉素处理后显著增加。OdhI基因的人工过表达导致2-酮戊二酸脱氢酶复合物的比活性降低，并且在没有任何触发的情况下增加谷氨酸产量。这些结果表明，OdhI的重新合成是青霉素诱导C.谷氨酸。此外，连续的谷氨酸生产实现了过度表达的odhI没有任何触发器。因此，odhI过表达的C.谷氨酸杆菌可用于有效的谷氨酸生产。
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. H-3-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.