Cargando…
Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis
BACKGROUND: Conversion of industrial processes to more nature-friendly modes is a crucial subject for achieving sustainable development. Utilization of hydrogen-oxidation reactions by hydrogenase as a driving force of bioprocess reaction can be an environmentally ideal method because the reaction cr...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3552938/ https://www.ncbi.nlm.nih.gov/pubmed/23305396 http://dx.doi.org/10.1186/1475-2859-12-2 |
_version_ | 1782256748489342976 |
---|---|
author | Oda, Takahiro Oda, Koji Yamamoto, Hiroaki Matsuyama, Akinobu Ishii, Masaharu Igarashi, Yasuo Nishihara, Hirofumi |
author_facet | Oda, Takahiro Oda, Koji Yamamoto, Hiroaki Matsuyama, Akinobu Ishii, Masaharu Igarashi, Yasuo Nishihara, Hirofumi |
author_sort | Oda, Takahiro |
collection | PubMed |
description | BACKGROUND: Conversion of industrial processes to more nature-friendly modes is a crucial subject for achieving sustainable development. Utilization of hydrogen-oxidation reactions by hydrogenase as a driving force of bioprocess reaction can be an environmentally ideal method because the reaction creates no pollutants. We expressed NAD-dependent alcohol dehydrogenase from Kluyveromyces lactis in a hydrogen-oxidizing bacterium: Ralstonia eutropha. This is the first report of hydrogen-driven in vivo coupling reaction of the alcohol dehydrogenase and indigenous soluble NAD-reducing hydrogenase. Asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol, which is a commercial building block for antibacterial agents, was performed using the transformant as the microbial cell catalyst. RESULTS: The two enzymes coupled in vitro in vials without a marked decrease of reactivity during the 20 hr reaction because of the hydrogenase reaction, which generates no by-product that affects enzymes. Alcohol dehydrogenase was expressed functionally in R. eutropha in an activity level equivalent to that of indigenous NAD-reducing hydrogenase under the hydrogenase promoter. The hydrogen-driven in vivo coupling reaction proceeded only by the transformant cell without exogenous addition of a cofactor. The decrease of reaction velocity at higher concentration of hydroxyacetone was markedly reduced by application of an in vivo coupling system. Production of (R)-1,2-propanediol (99.8% e.e.) reached 67.7 g/l in 76 hr with almost a constant rate using a jar fermenter. The reaction velocity under 10% P(H2) was almost equivalent to that under 100% hydrogen, indicating the availability of crude hydrogen gas from various sources. The in vivo coupling system enabled cell-recycling as catalysts. CONCLUSIONS: Asymmetric reduction of hydroxyacetone by a coupling reaction of the two enzymes continued in both in vitro and in vivo systems in the presence of hydrogen. The in vivo reaction system using R. eutropha transformant expressing heterologous alcohol dehydrogenase showed advantages for practical usage relative to the in vitro coupling system. The results suggest a hopeful perspective of the hydrogen-driven bioprocess as an environmentally outstanding method to achieve industrial green innovation. Hydrogen-oxidizing bacteria can be useful hosts for the development of hydrogen-driven microbial cell factories. |
format | Online Article Text |
id | pubmed-3552938 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-35529382013-01-28 Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis Oda, Takahiro Oda, Koji Yamamoto, Hiroaki Matsuyama, Akinobu Ishii, Masaharu Igarashi, Yasuo Nishihara, Hirofumi Microb Cell Fact Research BACKGROUND: Conversion of industrial processes to more nature-friendly modes is a crucial subject for achieving sustainable development. Utilization of hydrogen-oxidation reactions by hydrogenase as a driving force of bioprocess reaction can be an environmentally ideal method because the reaction creates no pollutants. We expressed NAD-dependent alcohol dehydrogenase from Kluyveromyces lactis in a hydrogen-oxidizing bacterium: Ralstonia eutropha. This is the first report of hydrogen-driven in vivo coupling reaction of the alcohol dehydrogenase and indigenous soluble NAD-reducing hydrogenase. Asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol, which is a commercial building block for antibacterial agents, was performed using the transformant as the microbial cell catalyst. RESULTS: The two enzymes coupled in vitro in vials without a marked decrease of reactivity during the 20 hr reaction because of the hydrogenase reaction, which generates no by-product that affects enzymes. Alcohol dehydrogenase was expressed functionally in R. eutropha in an activity level equivalent to that of indigenous NAD-reducing hydrogenase under the hydrogenase promoter. The hydrogen-driven in vivo coupling reaction proceeded only by the transformant cell without exogenous addition of a cofactor. The decrease of reaction velocity at higher concentration of hydroxyacetone was markedly reduced by application of an in vivo coupling system. Production of (R)-1,2-propanediol (99.8% e.e.) reached 67.7 g/l in 76 hr with almost a constant rate using a jar fermenter. The reaction velocity under 10% P(H2) was almost equivalent to that under 100% hydrogen, indicating the availability of crude hydrogen gas from various sources. The in vivo coupling system enabled cell-recycling as catalysts. CONCLUSIONS: Asymmetric reduction of hydroxyacetone by a coupling reaction of the two enzymes continued in both in vitro and in vivo systems in the presence of hydrogen. The in vivo reaction system using R. eutropha transformant expressing heterologous alcohol dehydrogenase showed advantages for practical usage relative to the in vitro coupling system. The results suggest a hopeful perspective of the hydrogen-driven bioprocess as an environmentally outstanding method to achieve industrial green innovation. Hydrogen-oxidizing bacteria can be useful hosts for the development of hydrogen-driven microbial cell factories. BioMed Central 2013-01-10 /pmc/articles/PMC3552938/ /pubmed/23305396 http://dx.doi.org/10.1186/1475-2859-12-2 Text en Copyright ©2013 Oda et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Oda, Takahiro Oda, Koji Yamamoto, Hiroaki Matsuyama, Akinobu Ishii, Masaharu Igarashi, Yasuo Nishihara, Hirofumi Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis |
title | Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis |
title_full | Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis |
title_fullStr | Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis |
title_full_unstemmed | Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis |
title_short | Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis |
title_sort | hydrogen-driven asymmetric reduction of hydroxyacetone to (r)-1,2-propanediol by ralstonia eutropha transformant expressing alcohol dehydrogenase from kluyveromyces lactis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3552938/ https://www.ncbi.nlm.nih.gov/pubmed/23305396 http://dx.doi.org/10.1186/1475-2859-12-2 |
work_keys_str_mv | AT odatakahiro hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis AT odakoji hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis AT yamamotohiroaki hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis AT matsuyamaakinobu hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis AT ishiimasaharu hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis AT igarashiyasuo hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis AT nishiharahirofumi hydrogendrivenasymmetricreductionofhydroxyacetonetor12propanediolbyralstoniaeutrophatransformantexpressingalcoholdehydrogenasefromkluyveromyceslactis |