Heterologous Biosynthesis of the Fungal Sesquiterpene Trichodermol in Saccharomyces cerevisiae

• Published in: Frontiers in microbiology Vol. 9; p. 1773
• Main Authors: Liu, Jianghua, Zhai, Yanan, Zhang, Yang, Zhu, Shuaiming, Liu, Gang, Che, Yongsheng
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 06.08.2018
• Subjects: FPP; heterologous biosynthesis; Microbiology; RNA-Seq; Saccharomyces cerevisiae; trichodermol; trichodiene; RNA-Seq; FPP; trichodiene; heterologous biosynthesis; qPCR; trichodermol; Saccharomyces cerevisiae
• ISSN: 1664-302X; 1664-302X
• DOI: 10.3389/fmicb.2018.01773

Trichodermol, a fungal sesquiterpene derived from the farnesyl diphosphate pathway, is the biosynthetic precursor for trichodermin, a member of the trichothecene class of fungal toxins produced mainly by the genera of and . Trichodermin is a promising candidate for the development of fungicides and antitumor agents due to its significant antifungal and cytotoxic effects. It can also serve as a scaffold to generate new congeners for structure-activity relationship (SAR) study. We reconstructed the biosynthetic pathway of trichodermol in BY4741, and investigated the effect of produced trichodermol on the host by RNA sequencing (RNA-Seq) and quantitative Real-time PCR analyses. Co-expression of pESC:: using plasmid pLLeu-tHMGR-UPC2.1 led to trichodiene production of 683 μg L , while integration of only the codon-optimized into the chromosome of yeast improved the production to 6,535 μg L . Subsequent expression of the codon-optimized cytochrome P450 monooxygenase encoding genes, and , resulted in trichodermol, with an estimated titer of 252 μg L at shake flask level. RNA-Seq and qPCR analyses revealed that the produced trichodermol downregulated the expression of the genes involved in ergosterol biosynthesis, but significantly upregulated the expression of related to membrane transport pathway in Collectively, we achieved the first heterologous biosynthesis of trichodermol by reconstructing its biosynthetic pathway in yeast, and the reconstructed pathway will serve as a platform to generate trichodermin analogs as potential candidates for agrochemicals and anticancer agents through further optimizations.