A Pseudomonas putida efflux pump acts on short-chain alcohols

Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle; Keasling, Jay

doi:10.1186/s13068-018-1133-9

Title: A Pseudomonas putida efflux pump acts on short-chain alcohols

Journal Article · Fri May 11 00:00:00 EDT 2018 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-018-1133-9· OSTI ID:1618742

Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle; Keasling, Jay

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.

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Research Organization:: Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1618742

Alternate ID(s):: OSTI ID: 1460319

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Vol. 11 Journal Issue: 1; ISSN 1754-6834

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: Netherlands

Language:: English

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Cited by: 29 works

Citation information provided by
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09 BIOMASS FUELS
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MOESM1 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254279 The current record is supplemented by this figure	figure	May 2018
MOESM1 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254279.v1 The current record is supplemented by this figure	figure	May 2018
MOESM2 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254297 The current record is supplemented by this figure	figure	May 2018
MOESM2 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254297.v1 The current record is supplemented by this figure	figure	May 2018
MOESM3 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254315.v1 The current record is supplemented by this figure	figure	May 2018
MOESM4 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254333 The current record is supplemented by this dataset	dataset	May 2018
MOESM4 of A Pseudomonas putida efflux pump acts on short-chain alcohols Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.6254333.v1 The current record is supplemented by this dataset	dataset	May 2018
A Pseudomonas putida efflux pump acts on short-chain alcohols [Supplemental Data] Basler, Georg; Thompson, Mitchell; Tullman-Ercek, Danielle https://doi.org/10.6084/m9.figshare.c.4097201.v1 The current record is supplemented by this collection	collection	May 2018