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Transcriptome analyses of the Corynebacterium glutamicum strains adapted to supraoptimal growth temperatures

ABSTRACT: C. glutamicum strains adapted to higher growth temperatures were obtained through an adaptive laboratory evolution experiment. To elucidate molecular basis for thermotolerance acquired by the evolved strains, we examined transcriptional responses of the evolved and parental strains to thermal stress using microarray technology.

ORGANISM(S): Corynebacterium glutamicum Corynebacterium glutamicum R

PROVIDER: GSE64654 | GEO | 2015/01/06

SECONDARY ACCESSION(S): PRJNA271577

REPOSITORIES: GEO

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Transcriptome analyses of the Corynebacterium glutamicum strains adapted to supraoptimal growth temperatures

Project description:C. glutamicum strains adapted to higher growth temperatures were obtained through an adaptive laboratory evolution experiment. To elucidate molecular basis for thermotolerance acquired by the evolved strains, we examined transcriptional responses of the evolved and parental strains to thermal stress using microarray technology. Each strain was grown at the optimal growth temperature (33 M-bM-^DM-^C). When OD610 reached around 7, the growth temperature increased to 41 M-bM-^DM-^C, and transcriptional changes of the evolved and parental strains were examined with microarray technology.

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Project description:Ciprofloxacin, an inhibitor of bacterial gyrase and topoisomerase IV, was shown to inhibit growth of C. glutamicum with concomitant excretion of L-glutamate. C. glutamicum strains overproducing L-lysine, L-arginine, L-ornithine, and putrescine, respectively, produced L-glutamate instead of the desired amino acid when exposed to ciprofloxacin. Even in the absence of the putative L-glutamate exporter gene yggB, ciprofloxacin effectively triggered L-glutamate production. When C. glutamicum wild type was cultivated under nitrogen-limiting conditions, 2-oxoglutarate rather than L-glutamate was produced as consequence of exposure to ciprofloxacin. Transcriptome analysis revealed that ciprofloxacin increased RNA levels of genes involved in DNA synthesis, repair and modification. Enzyme assays showed that 2-oxoglutarate dehydrogenase activity was decreased due to ciprofloxacin addition. Here, it was shown for the first time that production of L-glutamate by C. glutamicum may be triggered by an inhibitor of DNA synthesis and L-glutamate titers of up to 37 ± 1 mM and a substrate specific L-glutamate yield of 0.13 g/g were reached.

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