Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides
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 mod...
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2010
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paper:paper_13645072_v109_n2_p492_Nikel2023-06-08T16:11:44Z Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides Pettinari, María Julia Méndez, Beatriz Silvia Galvagno, Miguel Angel 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 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. Fil:Pettinari, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Méndez, B.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Galvagno, M.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2010 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13645072_v109_n2_p492_Nikel http://hdl.handle.net/20.500.12110/paper_13645072_v109_n2_p492_Nikel |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 Pettinari, María Julia Méndez, Beatriz Silvia Galvagno, Miguel Angel Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides |
| topic_facet |
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 |
| description |
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. |
| author |
Pettinari, María Julia Méndez, Beatriz Silvia Galvagno, Miguel Angel |
| author_facet |
Pettinari, María Julia Méndez, Beatriz Silvia Galvagno, Miguel Angel |
| author_sort |
Pettinari, María Julia |
| title |
Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides |
| title_short |
Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides |
| title_full |
Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides |
| title_fullStr |
Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides |
| title_full_unstemmed |
Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides |
| title_sort |
ethanol synthesis from glycerol by escherichia coli redox mutants expressing adhe from leuconostoc mesenteroides |
| publishDate |
2010 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13645072_v109_n2_p492_Nikel http://hdl.handle.net/20.500.12110/paper_13645072_v109_n2_p492_Nikel |
| work_keys_str_mv |
AT pettinarimariajulia ethanolsynthesisfromglycerolbyescherichiacoliredoxmutantsexpressingadhefromleuconostocmesenteroides AT mendezbeatrizsilvia ethanolsynthesisfromglycerolbyescherichiacoliredoxmutantsexpressingadhefromleuconostocmesenteroides AT galvagnomiguelangel ethanolsynthesisfromglycerolbyescherichiacoliredoxmutantsexpressingadhefromleuconostocmesenteroides |
| _version_ |
1768544463269920768 |