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Title: 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 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, highmore » 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.« less
Authors:
 [1] ;  [2] ;  [3] ;  [4]
  1. Univ. of California, Berkeley, CA (United States); Max Planck Institute for Molecular Plant Physiology, Potsdam (Germany)
  2. Univ. of California, Berkeley, CA (United States); Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  3. Northwestern Univ., Evanston, IL (United States)
  4. Univ. of California, Berkeley, CA (United States); Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of Denmark, Copenhagen (Denmark)
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 11; Journal Issue: 1; Related Information: © 2018 The Author(s).; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Energy Efficiency and Renewable Energy (EERE)
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; Higher alcohols; Short-chain alcohols; Next-generation biofuels; Efflux pumps; TtgABC; Pseudomonas putida; Tolerance; Toxicity
OSTI Identifier:
1460319

Basler, Georg, Thompson, Mitchell, Tullman-Ercek, Danielle, and Keasling, Jay. A Pseudomonas putida efflux pump acts on short-chain alcohols. United States: N. p., Web. doi:10.1186/s13068-018-1133-9.
Basler, Georg, Thompson, Mitchell, Tullman-Ercek, Danielle, & Keasling, Jay. A Pseudomonas putida efflux pump acts on short-chain alcohols. United States. doi:10.1186/s13068-018-1133-9.
Basler, Georg, Thompson, Mitchell, Tullman-Ercek, Danielle, and Keasling, Jay. 2018. "A Pseudomonas putida efflux pump acts on short-chain alcohols". United States. doi:10.1186/s13068-018-1133-9. https://www.osti.gov/servlets/purl/1460319.
@article{osti_1460319,
title = {A Pseudomonas putida efflux pump acts on short-chain alcohols},
author = {Basler, Georg and Thompson, Mitchell and Tullman-Ercek, Danielle and Keasling, Jay},
abstractNote = {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.},
doi = {10.1186/s13068-018-1133-9},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 11,
place = {United States},
year = {2018},
month = {5}
}

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