Sunday, July 26, 2009
P91
Adaptation of Escherichia coli for High Succinic Acid and Transcriptome Analysis Therefrom
Yeong-Deok Kwon and Pil Kim. Department of Biotechnology, Catholic University, 43-1 Yeokgok 2-dong, Wonmi-gu, Bucheon, 420-743, South Korea
To understand cell physiology against high succinic acid stress, a continuous culture was performed for 9 months using a wild E. coli against gradually increasing succinic acid and the transcriptome of the adapted E. coli for high succinate was analyzed.After 9 months operation of the continuous culture, we acquired the adapted E. coli which was able to grow in M9-glucose medium containing 1.3 M succinic acid. Total mRNA of the adapted E. coli and wild strain was compared using DNA microarray and further verified by real-time PCR. The adapted E. coli was able to grow without any lag phase in the M9-glucose medium containing 1.3 M succinate while wild W3110 showed more than 30 h lag phase. The cell growth of HST was about 10% higher than that of W3110 from 3 g/L glucose consumption.The microarray result showed up-regulation of 678 genes and down-regulation of 288 genes in the HST strain compared with wild strain.Genes involving osmoprotectant production were also up-regulated in the HST. The 12 % of the up-regulated genes were for the transport and binding proteins, mostly involved with the ABC transporter system.The 10.5 % of the up-regulated genes were for the"energy metabolism", particularly the genes involved in oxidative phosphorylation for ATP production. In the transcriptome analysis, HST strain significantly promoted the genes for production of increased osmolytes accumulation and many sugar transporters.The results from this work would provide strategy to modify cellular metabolism tolerating high metabolite concentration.
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