Identification and manipulation of a novel locus to improve cell tolerance to short-chain alcohols in <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Yan Chen Ying Wang Tian-Hua Chen Ming-Dong Yao Wen-Hai Xiao Bing-Zhi Li Ying-Jin Yuan |
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Institution: | 1.Key Laboratory of Systems Bioengineering (Ministry of Education),Tianjin University,Tianjin,People’s Republic of China;2.SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology,Tianjin University,Tianjin,People’s Republic of China |
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Abstract: | Escherichia coli KO11 is a popular ethanologenic strain, but is more sensitive to ethanol than other producers. Here, an ethanol-tolerant mutant EM was isolated from ultraviolet mutagenesis library of KO11. Comparative genomic analysis added by piecewise knockout strategy and complementation assay revealed EKO11_3023 (espA) within the 36.6-kb deletion from KO11 was the only locus responsible for ethanol sensitivity. Interestingly, when espA was deleted in strain W (the parent strain of KO11), ethanol tolerance was dramatically elevated to the level of espA-free hosts e.g., MG1655 and BL21(DE3)]. And overexpression of espA in strains MG1655 and BL21(DE3) led to significantly enhanced ethanol sensitivity. In addition to ethanol, deletion of espA also improved cell tolerance to other short-chain (C2–C4) alcohols, including methanol, isopropanol, n-butanol, isobutanol and 2-butanol. Therefore, espA was responsible for short-chain alcohol sensitivity of W-strains compared to other cells, which provides a potential engineering target for alcohols production. |
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