Metabolic engineering of Rhodotorula toruloides for resveratrol production

Microbial Cell Factories

Research

Qidou Gao¹  Zhumei Fang¹  Yijuan Liu¹  Xiaobing Yang¹  Mengyao Zhang²  Zongbao K. Zhao³  Zhiwei Gong⁴ 

[1] College of Enology, Northwest A&F University, Yangling, 712100, Xianyang, Shaanxi, China;College of Enology, Northwest A&F University, Yangling, 712100, Xianyang, Shaanxi, China;Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, 116023, Dalian, China;Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, 116023, Dalian, China;School of Chemistry and Chemical Engineering, Wuhan University of Science and Technology, 947 Heping Road, 430081, Wuhan, China;

关键词: Rhodotorula toruloides;    Resveratrol;    Metabolic engineering;    Cell factory;   

DOI  :  10.1186/s12934-022-02006-w

 received in 2022-11-02, accepted in 2022-12-17,  发布年份 2022

来源: Springer

【 摘 要 】

BackgroundResveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial production of resveratrol offers a promising alternative to plant- and chemical-based processes. The non-conventional oleaginous yeast Rhodotorula toruloides is a potential workhorse for the production of resveratrol that endowed with an efficient and intrinsic bifunctional phenylalanine/tyrosine ammonia-lyase (RtPAL) and malonyl-CoA pool, which may facilitate the resveratrol synthesis when properly rewired.ResultsResveratrol showed substantial stability and would not affect the R. toruloides growth during the yeast cultivation in flasks. The heterologus resveratrol biosynthesis pathway was established by introducing the 4-coumaroyl-CoA ligase (At4CL), and the stilbene synthase (VlSTS) from Arabidopsis thaliana and Vitis labrusca, respectively. Next, The resveratrol production was increased by 634% through employing the cinnamate-4-hydroxylase from A. thaliana (AtC4H), the fused protein At4CL::VlSTS, the cytochrome P450 reductase 2 from A. thaliana (AtATR2) and the endogenous cytochrome B5 of R. toruloides (RtCYB5). Then, the related endogenous pathways were optimized to affect a further 60% increase. Finally, the engineered strain produced a maximum titer of 125.2 mg/L resveratrol in YPD medium.ConclusionThe non-conventional oleaginous yeast R. toruloides was engineered for the first time to produce resveratrol. Protein fusion, co-factor channeling, and ARO4 and ARO7 overexpression were efficient for improving resveratrol production. The results demonstrated the potential of R. toruloides for resveratrol and other phenylpropanoids production.

【 授权许可】

CC BY   
© The Author(s) 2022

【 图 表 】

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