Re-engineering Escherichia coli for ethanol production.
A lactate producing derivative of Escherichia coli KO11, strain SZ110, was re-engineered for ethanol production by deleting genes encoding all fermentative routes for NADH and randomly inserting a promoterless mini-Tn5 cassette (transpososome) containing the complete Zymomonas mobilis ethanol pathway (pdc, adhA, and adhB) into the chromosome. By selecting for fermentative growth in mineral salts medium containing xylose, a highly productive strain was isolated in which the ethanol cassette had been integrated behind the rrlE promoter, designated strain LY160(KO11, Deltafrd::celY(Ec) DeltaadhE DeltaldhA, DeltaackA lacA::casAB(Ko) rrlE::(pdc( Zm)-adhA(Zm)-adhB(Zm)-FRT-rrlE)pflB(+)). This strain fermented 9% (w/v) xylose to 4% (w/v) ethanol in 48 h in mineral salts medium, nearly equal to the performance of KO11 with Luria broth.
Yomano
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Department Microbiology and Cell Science, University of Florida, Gainesville, FL 32611, USA.
York
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Zhou
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Shanmugam
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Ingram
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D001622:Betaine / Q000378:metabolism; D002849:Chromatography, Gas; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D005798:Genes, Bacterial; D005818:Genetic Engineering / Q000379:methods*; D019344:Lactic Acid / Q000378:metabolism; D009997:Osmotic Pressure; D016133:Polymerase Chain Reaction; D014994:Xylose / Q000378:metabolism
D003470:Culture Media; D019344:Lactic Acid; D000431:Ethanol; D001622:Betaine; D014994:Xylose