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Evolutionary engineering improves tolerance for replacement jet fuels in Saccharomyces cerevisiae

机译:进化工程提高了酿酒酵母替代喷气燃料的耐受性

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摘要

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 (tTcb3p¹⁻⁹⁸⁹), but not its complete deletion, was sufficient to recover the evolved phenotype improving limonene fitness 9-fold. tTcb3p¹⁻⁹⁸⁹ 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. tTcb3p¹⁻⁹⁸⁹ is the first example of successful engineering of phase tolerance and creates opportunities for production of the highly toxic C₁₀ alkenes in yeast.
机译:单萜是液态烃,其应用范围从风味和香气到替代喷气燃料。然而,它们的毒性对微生物合成提出了重大挑战。在这里,我们进化了耐柠檬烯的酿酒酵母菌株,并在200代进化时间过程中对6个菌株进行了测序。在traicalbin蛋白Tcb2p和Tcb3p中发现了突变。亲本菌株中的基因组重建表明,单个蛋白质(tTcb3p11)的截短而不是其完全缺失,足以恢复进化的表型,使柠檬烯适应性提高了9倍。 tTcb3p¹⁻⁹⁸⁹对其他两种单萜(β-ter烯和月桂烯)的耐受性分别提高了11倍和8倍,对生物喷气燃料混合物AMJ-700t(10%的异丙苯,50%的柠檬烯,40%的法呢烯)的耐受性4-折。 tTcb3p¹是成功进行相差工程的第一个例子,它为在酵母中生产高毒性的C₁₀烯烃创造了机会。

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