Creation of stress tolerant strain based on single nucleotide mutational analysis of in whole genome

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We performed experimental evolution under(5%) ethanol stress condition in Escherichia coli and six ethanol stress tolerant evolved strains were independently obtained. Transcriptomic analysis of evolved strains was performed. As a result of principal component analysis(PCA), gene expression in biosynthesis pathways of some amino acids were extracted and these are commonly up-regulated in the tolerant strains, suggesting that activating these pathways were involved in the achievement of ethanol tolerance. In support of this hypothesis, the supplementation of these amino acids to the culture medium increased the specific growth rate under ethanol stress. Mutational analysis was also performed by high-density oligonucleotide microarray and it was suggested that mutation does not significantly contribute to ethanol stress tolerance because there few common mutations among evolved strains.