Evolutionary Engineering Improves Tolerance for Replacement Jet Fuels in Saccharomyces cerevisiae

Applied and Environmental Microbiology
Timothy C R BrennanLars K Nielsen

Abstract

Monoterpenes are liquid hydrocarbons with applications ranging from flavor and fragrance to replacement jet fuel. Their toxicity, however, presents a major challenge for microbial synthesis. Here we evolved limonene-tolerant Saccharomyces cerevisiae strains and sequenced six strains across the 200-generation evolutionary time course. Mutations were found in the tricalbin proteins Tcb2p and Tcb3p. Genomic reconstruction in the parent strain showed that truncation of a single protein (tTcb3p(1-989)), but not its complete deletion, was sufficient to recover the evolved phenotype improving limonene fitness 9-fold. tTcb3p(1-989) increased tolerance toward two other monoterpenes (β-pinene and myrcene) 11- and 8-fold, respectively, and tolerance toward the biojet fuel blend AMJ-700t (10% cymene, 50% limonene, 40% farnesene) 4-fold. tTcb3p(1-989) is the first example of successful engineering of phase tolerance and creates opportunities for production of the highly toxic C10 alkenes in yeast.

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Citations

Sep 1, 2015·Current Opinion in Biotechnology·Kyle A Zingaro, Eleftherios Terry Papoutsakis
Mar 8, 2016·Trends in Biotechnology·Thomas C WilliamsIan T Paulsen
Feb 27, 2016·Applied Microbiology and Biotechnology·Esmer JongedijkJules Beekwilder
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Apr 6, 2018·Biotechnology for Biofuels·Stephanie A Davis LópezDanielle Tullman-Ercek
Nov 1, 2018·Biotechnology for Biofuels·Chun-Li LiuTaek Soon Lee
Dec 22, 2017·Applied Microbiology and Biotechnology·Yujin CaoHuizhou Liu
May 28, 2019·Applied Microbiology and Biotechnology·Sandra Moser, Harald Pichler
Dec 10, 2020·Biotechnology for Biofuels·Heiko Babel, Jens O Krömer
Nov 17, 2020·Frontiers in Bioengineering and Biotechnology·Laura Ellen Walls, Leonardo Rios-Solis
Feb 28, 2021·Cell·Yuzhong LiuJay D Keasling
Sep 4, 2020·Biotechnology Advances·Yuyao RenYongjin J Zhou
Jul 15, 2021·Critical Reviews in Biotechnology·Chun-Li LiuTianwei Tan
Dec 24, 2019·Journal of Agricultural and Food Chemistry·Xixian ChenNicholas D Lindley

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