A combined strategy for the overproduction of complex ergot alkaloid agroclavine

Zhi-Pu Yu; Chunyan An; Yongpeng Yao; Chang-Yun Wang; Zhoutong Sun; Chengsen Cui; Ling Liu; Shu-Shan Gao

doi:10.1016/j.synbio.2022.08.003

A combined strategy for the overproduction of complex ergot alkaloid agroclavine

Synth Syst Biotechnol. 2022 Aug 20;7(4):1126-1132. doi: 10.1016/j.synbio.2022.08.003. eCollection 2022 Dec.

Authors

Zhi-Pu Yu^{1

2}, Chunyan An³, Yongpeng Yao⁴, Chang-Yun Wang^{1

2}, Zhoutong Sun^{5

6}, Chengsen Cui^{5

6}, Ling Liu⁴, Shu-Shan Gao^{5

6}

Affiliations

¹ Key Laboratory of Marine Drugs, The Ministry of Education of China, Institute of Evolution & Marine Biodiversity, School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, PR China.
² Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266003, PR China.
³ Beijing Institute for Drug Control, NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine, Beijing, 102206, PR China.
⁴ State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, PR China.
⁵ Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, PR China.
⁶ National Technology Innovation Center of Synthetic Biology, Tianjin, 300308, PR China.

Abstract

Microbial cell factories (MCFs) and cell-free systems (CFSs) are generally considered as two unrelated approaches for the biosynthesis of biomolecules. In the current study, two systems were combined together for the overproduction of agroclavine (AC), a structurally complex ergot alkaloid. The whole biosynthetic pathway for AC was split into the early pathway and the late pathway at the point of the FAD-linked oxidoreductase EasE, which was reconstituted in an MCF (Aspergillus nidulans) and a four-enzyme CFS, respectively. The final titer of AC of this combined system is 1209 mg/L, which is the highest one that has been reported so far, to the best of our knowledge. The development of such a combined route could potentially avoid the limitations of both MCF and CFS systems, and boost the production of complex ergot alkaloids with polycyclic ring systems.

Keywords: Agroclavine; Cell-free systems; Ergot alkaloids; Microbial cell factories; Overproduction.