Increased ethanol resistance in <Emphasis Type="Italic">Ethanolic Escherichia coli</Emphasis> by Insertion of heat-shock genes BEM1 and SOD2 from <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> |
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Authors: | Soo Jin Lee Eun Kyoung Oh Young Hoon Oh Jong In Won Sung Ok Han Jin Won Lee |
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Institution: | (1) National Center for Agricultural Utilization Research, Agricultural Research Service, USDA, 1815 North University Street, Peoria, IL 61604, USA;(2) Institute for Microbial and Biochemical Technology, Forest Service, Forest Products Laboratory, USDA, One Gifford Pinchot Drive, Madison, WI 53726–2398, USA |
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Abstract: | Ethanol is generally toxic to microorganisms, and intracellular and extracellular accumulation of ethanol inhibits cell growth
and metabolism. In this study, pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB) were cloned into pET-32a vector
and then introduced into E. coli BL21 to produce ethanol. Heat shock genes (BEM1 and SOD2) from Saccharomyces cerevisiae were inserted into recombinant ethanolic E. coli using pET28_a vector to improve ethanol shock resistance. Three different strains were constructed: Ethanolic E. coli (adhB and pdc genes inserted using pET32_a vector), BEM1 gene-inserted E. coli (BEM1 inserted using pET_28a), and SOD2-inserted E. coli (SOD2 inserted using pET28_a). Construction of these three different strains allowed comparison of the functions of these
heat shock genes as well as their roles in ethanol tolerance. The toxicity of ethanol in recombinant ethanolic E. coli was tested by measuring cell growth in response to various ethanol concentrations. The results show that SOD2-inserted E. coli showed higher ethanol resistance than ethanolic E. coli. |
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