Reconstitution of biosynthetic pathway for mushroom-derived cyathane diterpenes in yeast and generation of new "non-natural" analogues

Ke Ma; Yuting Zhang; Cui Guo; Yanlong Yang; Junjie Han; Bo Yu; Wenbing Yin; Hongwei Liu

doi:10.1016/j.apsb.2021.04.014

Reconstitution of biosynthetic pathway for mushroom-derived cyathane diterpenes in yeast and generation of new "non-natural" analogues

Acta Pharm Sin B. 2021 Sep;11(9):2945-2956. doi: 10.1016/j.apsb.2021.04.014. Epub 2021 May 3.

Authors

Ke Ma^{1

2}, Yuting Zhang^{1

2}, Cui Guo^{1

2}, Yanlong Yang¹, Junjie Han¹, Bo Yu^{1

3}, Wenbing Yin^{1

2}, Hongwei Liu^{1

2}

Affiliations

¹ State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
² Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, China.
³ CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.

Abstract

Mushroom-derived cyathane-type diterpenes possess unusual chemical skeleton and diverse bioactivities. To efficiently supply bioactive cyathanes for deep studies and explore their structural diversity, de novo synthesis of cyathane diterpenes in a geranylgeranyl pyrophosphate engineered Saccharomyces cerevisiae is investigated. Aided by homologous analyses, one new unclustered FAD-dependent oxidase EriM accounting for the formation of allyl aldehyde and three new NADP(H)-dependent reductases in the biosynthesis of cyathanes are identified and elucidated. By combinatorial biosynthetic strategy, S. cerevisiae strains generating twenty-two cyathane-type diterpenes, including seven "unnatural" cyathane xylosides (12, 13, 14a, 14b, 19, 20, and 22) are established. Compounds 12-14, 19, and 20 show significant neurotrophic effects on PC12 cells in the dose of 6.3-25.0 μmol/L. These studies provide new insights into the divergent biosynthesis of mushroom-originated cyathanes and a straightforward approach to produce bioactive cyathane-type diterpenes.

Keywords: Biosynthesis; Cyathane-type diterpene; Heterologous expression; Non-enzymatic reaction.