, Volume 30, Issue 12, pp 2097-2103
Date: 05 Sep 2008

Re-engineering Escherichia coli for ethanol production

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Abstract

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, Δfrd::celY Ec ΔadhE ΔldhA, ΔackA 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.