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A Pseudomonas putida efflux pump acts on short-chain alcohols

BACKGROUND: The microbial production of biofuels is complicated by a tradeoff between yield and toxicity of many fuels. Efflux pumps enable bacteria to tolerate toxic substances by their removal from the cells while bypassing the periplasm. Their use for the microbial production of biofuels can help...

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Autores principales: Basler, Georg, Thompson, Mitchell, Tullman-Ercek, Danielle, Keasling, Jay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5946390/
https://www.ncbi.nlm.nih.gov/pubmed/29760777
http://dx.doi.org/10.1186/s13068-018-1133-9
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author Basler, Georg
Thompson, Mitchell
Tullman-Ercek, Danielle
Keasling, Jay
author_facet Basler, Georg
Thompson, Mitchell
Tullman-Ercek, Danielle
Keasling, Jay
author_sort Basler, Georg
collection PubMed
description BACKGROUND: The microbial production of biofuels is complicated by a tradeoff between yield and toxicity of many fuels. Efflux pumps enable bacteria to tolerate toxic substances by their removal from the cells while bypassing the periplasm. Their use for the microbial production of biofuels can help to improve cell survival, product recovery, and productivity. However, no native efflux pump is known to act on the class of short-chain alcohols, important next-generation biofuels, and it was considered unlikely that such an efflux pump exists. RESULTS: We report that controlled expression of the RND-type efflux pump TtgABC from Pseudomonas putida DOT-T1E strongly improved cell survival in highly toxic levels of the next-generation biofuels n-butanol, isobutanol, isoprenol, and isopentanol. GC-FID measurements indicated active efflux of n-butanol when the pump is expressed. Conversely, pump expression did not lead to faster growth in media supplemented with low concentrations of n-butanol and isopentanol. CONCLUSIONS: TtgABC is the first native efflux pump shown to act on multiple short-chain alcohols. Its controlled expression can be used to improve cell survival and increase production of biofuels as an orthogonal approach to metabolic engineering. Together with the increased interest in P. putida for metabolic engineering due to its flexible metabolism, high native tolerance to toxic substances, and various applications of engineering its metabolism, our findings endorse the strain as an excellent biocatalyst for the high-yield production of next-generation biofuels. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1133-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-59463902018-05-14 A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg Thompson, Mitchell Tullman-Ercek, Danielle Keasling, Jay Biotechnol Biofuels Research BACKGROUND: The microbial production of biofuels is complicated by a tradeoff between yield and toxicity of many fuels. Efflux pumps enable bacteria to tolerate toxic substances by their removal from the cells while bypassing the periplasm. Their use for the microbial production of biofuels can help to improve cell survival, product recovery, and productivity. However, no native efflux pump is known to act on the class of short-chain alcohols, important next-generation biofuels, and it was considered unlikely that such an efflux pump exists. RESULTS: We report that controlled expression of the RND-type efflux pump TtgABC from Pseudomonas putida DOT-T1E strongly improved cell survival in highly toxic levels of the next-generation biofuels n-butanol, isobutanol, isoprenol, and isopentanol. GC-FID measurements indicated active efflux of n-butanol when the pump is expressed. Conversely, pump expression did not lead to faster growth in media supplemented with low concentrations of n-butanol and isopentanol. CONCLUSIONS: TtgABC is the first native efflux pump shown to act on multiple short-chain alcohols. Its controlled expression can be used to improve cell survival and increase production of biofuels as an orthogonal approach to metabolic engineering. Together with the increased interest in P. putida for metabolic engineering due to its flexible metabolism, high native tolerance to toxic substances, and various applications of engineering its metabolism, our findings endorse the strain as an excellent biocatalyst for the high-yield production of next-generation biofuels. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1133-9) contains supplementary material, which is available to authorized users. BioMed Central 2018-05-11 /pmc/articles/PMC5946390/ /pubmed/29760777 http://dx.doi.org/10.1186/s13068-018-1133-9 Text en © The Author(s) 2018 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
Basler, Georg
Thompson, Mitchell
Tullman-Ercek, Danielle
Keasling, Jay
A Pseudomonas putida efflux pump acts on short-chain alcohols
title A Pseudomonas putida efflux pump acts on short-chain alcohols
title_full A Pseudomonas putida efflux pump acts on short-chain alcohols
title_fullStr A Pseudomonas putida efflux pump acts on short-chain alcohols
title_full_unstemmed A Pseudomonas putida efflux pump acts on short-chain alcohols
title_short A Pseudomonas putida efflux pump acts on short-chain alcohols
title_sort pseudomonas putida efflux pump acts on short-chain alcohols
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5946390/
https://www.ncbi.nlm.nih.gov/pubmed/29760777
http://dx.doi.org/10.1186/s13068-018-1133-9
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