Stabilization of pet operon plasmids and ethanol production in Escherichia coli strains lacking lactate dehydrogenase and pyruvate formate-lyase activities.

Applied and Environmental Microbiology
R B HespellR J Bothast

Abstract

In the last decade, a major goal of research in biofuels has been to metabolically engineer microorganisms to ferment multiple sugars from biomass or agricultural wastes to fuel ethanol. Escherichia coli strains genetically engineered to contain the pet operon (Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase B genes) produce high levels of ethanol. Strains carrying the pet operon in plasmid (e.g., E. coli B/pLOI297) or in chromosomal (e.g., E. coli KO11) sites require antibiotics in the media to maintain genetic stability and high ethanol productivity. To overcome this requirement, we used the conditionally lethal E. coli strain FMJ39, which carries mutations for lactate dehydrogenase and pyruvate formate lyase and grows aerobically but is incapable of anaerobic growth unless these mutations are complemented. E. coli FBR1 and FBR2 were created by transforming E. coli FMJ39 with the pet operon plasmids pLOI295 and pLOI297, respectively. Both strains were capable of anaerobic growth and displayed no apparent pet plasmid losses after 60 generations in serially transferred (nine times) anaerobic batch cultures. In contrast, similar aerobic cultures rapidly lost plasmids. In high-cell-density batch fermentations, ...Continue Reading

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Citations

Sep 24, 2005·Journal of Molecular Microbiology and Biotechnology·Masayuki InuiHideaki Yukawa
May 9, 2006·Journal of Industrial Microbiology & Biotechnology·Gregory J O MartinNeville B Pamment
Oct 16, 2007·Biotechnology Letters·Consuelo Vázquez-LimónJoel Osuna
Apr 2, 2008·Annals of the New York Academy of Sciences·Joy Doran Peterson, Lonnie O Ingram
Jul 13, 2000·Critical Reviews in Biotechnology·J C Diaz Ricci, M E Hernández
May 15, 2008·The Plant Journal : for Cell and Molecular Biology·Joy Doran-PetersonSarah K Brandon
Feb 13, 2007·Applied Microbiology and Biotechnology·Bärbel Hahn-HägerdalMarie F Gorwa-Grauslund
Dec 18, 2020·Frontiers in Bioengineering and Biotechnology·Ju-Jiun PangSi-Yu Li

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