A Pseudomonas putida efflux pump acts on short-chain alcohols

Biotechnology for Biofuels
Georg BaslerJay D Keasling

Abstract

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. 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. 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...Continue Reading

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Citations

Apr 17, 2020·Biotechnology Journal·Ying ZhuChun Li
Apr 8, 2020·Frontiers in Microbiology·Wing-Jin LiNick Wierckx
May 7, 2020·Biotechnology and Bioengineering·Sofia FerreiraIsabel Rocha
Aug 23, 2020·Applied and Environmental Microbiology·Mitchell G ThompsonJay D Keasling
Aug 18, 2020·Applied and Environmental Microbiology·Peng LiuYu-Guo Zheng
Nov 24, 2020·Frontiers in Bioengineering and Biotechnology·Maryna VasylkivskaPetra Patakova
Jan 8, 2021·Applied Microbiology and Biotechnology·Barbora BranskaPetra Patakova
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Jun 27, 2021·Nature Reviews. Microbiology·Jay KeaslingEric Sundstrom
Jul 6, 2021·Essays in Biochemistry·Nora Lisa BitzenhoferAnita Loeschcke

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Methods Mentioned

BETA
PCR
flow cytometry

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