Synthesis of cembratriene-ol and cembratriene-diol in yeast via the MVA pathway

Microbial Cell Factories

Michael P. Timko¹  Ning Fang²  Yu Zhang²  Xiaofeng Liu²  Zhongfeng Zhang²  Chunkai Wang²  Yanhua Liu²  Hongbo Zhang²  Yongmei Du²  Shiquan Bian² 

[1]Department of Biology, University of Virginia, 485 McCormick Road, 22904, Charlottesville, VA, USA

[2]TRI of CAAS-UVA Joint Laboratory of Synthetic Biology, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, 266101, Qingdao, China

关键词: Cembranoids;    Tobacco;    Biosynthesis;    Yeast;    CBTS1;    CYP450;    MVA pathway;   

DOI  :  10.1186/s12934-021-01523-4

来源: Springer

【 摘 要 】

BackgroundCembranoids are one kind of diterpenoids with multiple biological activities. The tobacco cembratriene-ol (CBT-ol) and cembratriene-diol (CBT-diol) have high anti-insect and anti-fungal activities, which is attracting great attentions for their potential usage in sustainable agriculture. Cembranoids were supposed to be formed through the 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway, yet the involvement of mevalonate (MVA) pathway in their synthesis remains unclear. Exploring the roles of MVA pathway in cembranoid synthesis could contribute not only to the technical approach but also to the molecular mechanism for cembranoid biosynthesis.ResultsWe constructed vectors to express cembratriene-ol synthase (CBTS1) and its fusion protein (AD-CBTS1) containing an N-terminal GAL4 AD domain as a translation leader in yeast. Eventually, the modified enzyme AD-CBTS1 was successfully expressed, which further resulted in the production of CBT-ol in the yeast strain BY-T20 with enhanced MVA pathway for geranylgeranyl diphosphate (GGPP) production but not in other yeast strains with low GGPP supply. Subsequently, CBT-diol was also synthesized by co-expression of the modified enzyme AD-CBTS1 and BD-CYP450 in the yeast strain BY-T20.ConclusionsWe demonstrated that yeast is insensitive to the tobacco anti-fungal compound CBT-ol or CBT-diol and could be applied to their biosynthesis. This study further established a feasibility for cembranoid production via the MVA pathway and provided an alternative bio-approach for cembranoid biosynthesis in microbes.

【 授权许可】

CC BY