Engineered Saccharomyces cerevisiae for de novo δ-tocotrienol biosynthesis

Abstract

As a vitamin E isomer, δ-tocotrienol has attracted much attention owing to its rich biological activities for human health, especially anticancer activities. Microbial biosynthesis constitutes a promising strategy of δ-tocotrienol production owing to its economic and environmental advantages. In this study, we employed modular engineering strategies to reconstruct and optimize a de novo δ-tocotrienol biosynthesis pathway in Saccharomyces cerevisiae. Subsequently, the rate-limiting steps were identified and eliminated and the key enzymes were assembled in the δ-tocotrienol biosynthesis module to develop a substrate channeling and improve their catalytic efficiency. Furthermore, the shikimate and δ-tocotrienol biosynthesis modules were optimized via combination strategies to further increase δ-tocotrienol production, following which the δ-tocotrienol titer reached 1455.5 μg/L. Finally, overexpression of the endogenous transporter PDR11 and two-phase extraction fermentation were employed for δ-tocotrienol production, yielding up to 3262.2 μg/L δ-tocotrienol. Thus, the findings of this study demonstrate the possibility of efficient δ-tocotrienol biosynthesis in S. cerevisiae.

Keywords

δ-Tocotrienol / Saccharomyces cerevisiae / De novo biosynthesis / Substrate channeling / Two-phase extraction fermentation

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Funding

Key Research and Development Program of China(2018YFA0900300); National Natural Science Foundation of China(32021005); Fundamental Research Funds for the Central Universities(JUSRP622004)