Abstract:
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.
Registro:
Documento: |
Artículo
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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
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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 |
Año: | 2010
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Volumen: | 109
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Número: | 2
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Página de inicio: | 492
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Página de fin: | 504
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DOI: |
http://dx.doi.org/10.1111/j.1365-2672.2010.04668.x |
Título revista: | Journal of Applied Microbiology
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Título revista abreviado: | J. Appl. Microbiol.
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ISSN: | 13645072
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CODEN: | JAMIF
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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, 1.1.1.1; Aldehyde Oxidoreductases, 1.2.-; Ethanol, 64-17-5; Glycerol, 56-81-5; acetaldehyde dehydrogenase, 1.2.1.5
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PDF: | https://bibliotecadigital.exactas.uba.ar/download/paper/paper_13645072_v109_n2_p492_Nikel.pdf |
Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13645072_v109_n2_p492_Nikel |
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Citas:
---------- 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.
http://dx.doi.org/10.1111/j.1365-2672.2010.04668.x---------- 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.
http://dx.doi.org/10.1111/j.1365-2672.2010.04668.x---------- 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.
http://dx.doi.org/10.1111/j.1365-2672.2010.04668.x---------- 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.
http://dx.doi.org/10.1111/j.1365-2672.2010.04668.x