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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.

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Acknowledgements

This research was supported by grants from the U.S. Department of Agriculture (01-35504-10669 and 00-52104-9704), the U.S. Department of Energy (FG02-96ER20222 and FG36-04GO14019), and by the Verenium Corporation.

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Authors and Affiliations

  1. Department Microbiology and Cell Science, University of Florida, Gainesville, FL, 32611, USA

    L. P. Yomano, S. W. York, S. Zhou, K. T. Shanmugam & L. O. Ingram

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  1. L. P. Yomano
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  2. S. W. York
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  3. S. Zhou
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  4. K. T. Shanmugam
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  5. L. O. Ingram
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Correspondence to L. O. Ingram.

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Yomano, L.P., York, S.W., Zhou, S. et al. Re-engineering Escherichia coli for ethanol production. Biotechnol Lett 30, 2097–2103 (2008). https://doi.org/10.1007/s10529-008-9821-3

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  • DOI: https://doi.org/10.1007/s10529-008-9821-3

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