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Corynebacterium glutamicum is known for its ability to produce glutamic acid and has been utilized for the fermentative production of various amino acids. Glutamic acid production in C. glutamicum is induced by penicillin. In this study, we analyzed the transcriptome and metabolome of C. glutamicum to understand the mechanism of penicillin‐induced glutamic acid production. Transcriptomic analysis with DNA microarray revealed that expression of some glycolysis‐ and TCA cycle‐related genes, which include those encoding the enzymes involved in conversion of glucose to 2‐oxoglutaric acid, was upregulated after penicillin addition. Meanwhile, expression of some TCA cycle‐related genes, encoding the enzymes for conversion of 2‐oxoglutaric acid to oxaloacetic acid, and the anaplerotic reactions decreased. In addition, expression of NCgl1221 and odhI, encoding proteins involved in glutamic acid excretion and inhibition of the 2‐oxoglutarate dehydrogenase, respectively, was upregulated. Functional category enrichment analysis of genes upregulated and downregulated after penicillin addition revealed that genes for signal transduction systems were enriched among upregulated genes, whereas those for energy production and carbohydrate and amino acid metabolisms were enriched among the downregulated genes. As for the metabolomic analysis using capillary electrophoresis time‐of‐flight mass spectrometry, the intracellular content of most metabolites of the glycolysis and the TCA cycle decreased dramatically after penicillin addition. Overall, these results indicate that the cellular metabolism and glutamic acid excretion are mainly optimized at the transcription level during penicillin‐induced glutamic acid production by C. glutamicum.
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