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Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol

BACKGROUND: Rhodopseudomonas palustris is a versatile microbe that encounters an innate redox imbalance while growing photoheterotrophically with reduced substrates. The resulting excess in reducing equivalents, together with ATP from photosynthesis, could be utilized to drive a wide range of biocon...

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Autores principales: Doud, Devin F. R., Holmes, Eric C., Richter, Hanno, Molitor, Bastian, Jander, Georg, Angenent, Largus T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5504763/
https://www.ncbi.nlm.nih.gov/pubmed/28702083
http://dx.doi.org/10.1186/s13068-017-0864-3
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author Doud, Devin F. R.
Holmes, Eric C.
Richter, Hanno
Molitor, Bastian
Jander, Georg
Angenent, Largus T.
author_facet Doud, Devin F. R.
Holmes, Eric C.
Richter, Hanno
Molitor, Bastian
Jander, Georg
Angenent, Largus T.
author_sort Doud, Devin F. R.
collection PubMed
description BACKGROUND: Rhodopseudomonas palustris is a versatile microbe that encounters an innate redox imbalance while growing photoheterotrophically with reduced substrates. The resulting excess in reducing equivalents, together with ATP from photosynthesis, could be utilized to drive a wide range of bioconversions. The objective of this study was to genetically modify R. palustris to provide a pathway to reduce n-butyrate into n-butanol for maintaining redox balance. RESULTS: Here, we constructed and expressed a plasmid-based pathway for n-butanol production from Clostridium acetobutylicum ATCC 824 in R. palustris. We maintained the environmental conditions in such a way that this pathway functioned as the obligate route to re-oxidize excess reducing equivalents, resulting in an innate selection pressure. The engineered strain of R. palustris grew under otherwise restrictive redox conditions and achieved concentrations of 1.5 mM n-butanol at a production rate of 0.03 g L(−1) day(−1) and a selectivity (i.e., products compared to the consumed substrate) of close to 40%. Since the theoretical maximum selectivity is 45%, the engineered strain converted close to its maximum selectivity. CONCLUSIONS: The innate redox imbalance of R. palustris can be used to drive the reduction of n-butyrate into n-butanol after expression of a plasmid-based enzyme from a butanol-producing Clostridium strain. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-017-0864-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-55047632017-07-12 Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol Doud, Devin F. R. Holmes, Eric C. Richter, Hanno Molitor, Bastian Jander, Georg Angenent, Largus T. Biotechnol Biofuels Research BACKGROUND: Rhodopseudomonas palustris is a versatile microbe that encounters an innate redox imbalance while growing photoheterotrophically with reduced substrates. The resulting excess in reducing equivalents, together with ATP from photosynthesis, could be utilized to drive a wide range of bioconversions. The objective of this study was to genetically modify R. palustris to provide a pathway to reduce n-butyrate into n-butanol for maintaining redox balance. RESULTS: Here, we constructed and expressed a plasmid-based pathway for n-butanol production from Clostridium acetobutylicum ATCC 824 in R. palustris. We maintained the environmental conditions in such a way that this pathway functioned as the obligate route to re-oxidize excess reducing equivalents, resulting in an innate selection pressure. The engineered strain of R. palustris grew under otherwise restrictive redox conditions and achieved concentrations of 1.5 mM n-butanol at a production rate of 0.03 g L(−1) day(−1) and a selectivity (i.e., products compared to the consumed substrate) of close to 40%. Since the theoretical maximum selectivity is 45%, the engineered strain converted close to its maximum selectivity. CONCLUSIONS: The innate redox imbalance of R. palustris can be used to drive the reduction of n-butyrate into n-butanol after expression of a plasmid-based enzyme from a butanol-producing Clostridium strain. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-017-0864-3) contains supplementary material, which is available to authorized users. BioMed Central 2017-07-11 /pmc/articles/PMC5504763/ /pubmed/28702083 http://dx.doi.org/10.1186/s13068-017-0864-3 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Doud, Devin F. R.
Holmes, Eric C.
Richter, Hanno
Molitor, Bastian
Jander, Georg
Angenent, Largus T.
Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
title Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
title_full Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
title_fullStr Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
title_full_unstemmed Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
title_short Metabolic engineering of Rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
title_sort metabolic engineering of rhodopseudomonas palustris for the obligate reduction of n-butyrate to n-butanol
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5504763/
https://www.ncbi.nlm.nih.gov/pubmed/28702083
http://dx.doi.org/10.1186/s13068-017-0864-3
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