Nikel, P.I.; Ramirez, M.C.; Pettinari, M.J.; Méndez, B.S.; Galvagno, M.A. "Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides" (2010) Journal of Applied Microbiology. 109(2):492-504
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Aims: Analysis of the physiology and metabolism of Escherichia coli arcA and creC mutants expressing a bifunctional alcohol-acetaldehyde dehydrogenase from Leuconostoc mesenteroides growing on glycerol under oxygen-restricted conditions. The effect of an ldhA mutation and different growth medium modifications was also assessed. Methods and Results: Expression of adhE in E. coli CT1061 [arcA creC(Con)] resulted in a 1·4-fold enhancement in ethanol synthesis. Significant amounts of lactate were produced during micro-oxic cultures and strain CT1061LE, in which fermentative lactate dehydrogenase was deleted, produced up to 6·5 ± 0·3 g l-1 ethanol in 48 h. Escherichia coli CT1061LE derivatives resistant to >25 g l-1 ethanol were obtained by metabolic evolution. Pyruvate and acetaldehyde addition significantly increased both biomass and ethanol concentrations, probably by overcoming acetyl-coenzyme A (CoA) shortage. Yeast extract also promoted growth and ethanol synthesis, and this positive effect was mainly attributable to its vitamin content. Two-stage bioreactor cultures were conducted in a minimal medium containing 100 μg l-1 calcium d-pantothenate to evaluate oxic acetyl-CoA synthesis followed by a switch into fermentative conditions. Ethanol reached 15·4 ± 0·9 g l-1 with a volumetric productivity of 0·34 ± 0·02 g l-1 h-1. Conclusions: Escherichia coli responded to adhE over-expression by funnelling carbon and reducing equivalents into a highly reduced metabolite, ethanol. Acetyl-CoA played a key role in micro-oxic ethanol synthesis and growth. Significance and Impact of the Study: Insight into the micro-oxic metabolism of E. coli growing on glycerol is essential for the development of efficient industrial processes for reduced biochemicals production from this substrate, with special relevance to biofuels synthesis. © 2010 The Society for Applied Microbiology.


Documento: Artículo
Título:Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides
Autor:Nikel, P.I.; Ramirez, M.C.; Pettinari, M.J.; Méndez, B.S.; Galvagno, M.A.
Filiación:Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín, Colectora Av. General Paz 5445, (B1650KNA) San Martín, Buenos Aires, Argentina
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Ciudad Autonoma de Buenos Aires, Argentina
Departamento de Ingeniería Química, Facultad de Ingeniería, Ciudad Autonoma de Buenos Aires, Argentina
Palabras clave:acetyl-CoA availability; alcohol-acetaldehyde dehydrogenase; Escherichia coli; glycerol metabolism; heterologous gene expression; microaerobiosis; microbial physiology; acetyl coenzyme A; alcohol; glycerol; lactate dehydrogenase; acetaldehyde; bacterium; biomass; bioreactor; carbon; developmental biology; enzyme activity; ethanol; fermentation; gene expression; industrial production; metabolism; metabolite; mutation; redox potential; vitamin; yeast; aerobic metabolism; article; bacterial strain; biomass; biosynthesis; Escherichia coli; fermentation; gene expression; Leuconostoc mesenteroides; nonhuman; Acetyl Coenzyme A; Alcohol Dehydrogenase; Aldehyde Oxidoreductases; Escherichia coli; Ethanol; Glycerol; Leuconostoc; Mutation; Oxidation-Reduction; Arca; Escherichia coli; Leuconostoc mesenteroides
Página de inicio:492
Página de fin:504
Título revista:Journal of Applied Microbiology
Título revista abreviado:J. Appl. Microbiol.
CAS:acetyl coenzyme A, 72-89-9; alcohol, 64-17-5; glycerol, 56-81-5; lactate dehydrogenase, 9001-60-9; Acetyl Coenzyme A, 72-89-9; Alcohol Dehydrogenase,; Aldehyde Oxidoreductases, 1.2.-; Ethanol, 64-17-5; Glycerol, 56-81-5; acetaldehyde dehydrogenase,


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---------- APA ----------
Nikel, P.I., Ramirez, M.C., Pettinari, M.J., Méndez, B.S. & Galvagno, M.A. (2010) . Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides. Journal of Applied Microbiology, 109(2), 492-504.
---------- CHICAGO ----------
Nikel, P.I., Ramirez, M.C., Pettinari, M.J., Méndez, B.S., Galvagno, M.A. "Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides" . Journal of Applied Microbiology 109, no. 2 (2010) : 492-504.
---------- MLA ----------
Nikel, P.I., Ramirez, M.C., Pettinari, M.J., Méndez, B.S., Galvagno, M.A. "Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides" . Journal of Applied Microbiology, vol. 109, no. 2, 2010, pp. 492-504.
---------- VANCOUVER ----------
Nikel, P.I., Ramirez, M.C., Pettinari, M.J., Méndez, B.S., Galvagno, M.A. Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides. J. Appl. Microbiol. 2010;109(2):492-504.