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 C 10 alkenes in yeast.
CITATION STYLE
Brennan, T. C. R., Williams, T. C., Schulz, B. L., Palfreyman, R. W., Krömer, J. O., & Nielsen, L. K. (2015). Evolutionary engineering improves tolerance for replacement jet fuels in Saccharomyces cerevisiae. Applied and Environmental Microbiology, 81(10), 3316–3325. https://doi.org/10.1128/AEM.04144-14
Mendeley helps you to discover research relevant for your work.