Gene identification and semisynthesis of the anti-inflammatory oleanane-type triterpenoid wilforlide A

Bibliographische Detailangaben
Veröffentlicht in:	The New phytologist. - 1979. - 241(2024), 4 vom: 01. Jan., Seite 1720-1731
1. Verfasser:	Liu, Yuan (VerfasserIn)
Weitere Verfasser:	Zhou, Jiawei, Liu, Panting, Hu, Tianyuan, Liu, Xuan, Gao, Jie, Ma, Lin, Lu, Yun, Li, Dan, Jiang, Zhouqian, Zhang, Xianan, Huang, Luqi, Gao, Wei, Wu, Xiaoyi, Zhang, Yifeng, Liu, Changli
Format:	Online-Aufsatz
Sprache:	English
Veröffentlicht:	2024
Zugriff auf das übergeordnete Werk:	The New phytologist
Schlagworte:	Journal Article Tripterygium wilfordii abrusgenic acid biosynthesis cytochrome P450 triterpenoids wilforlide A 84104-71-2 oleanane Triterpenes mehr... Anti-Inflammatory Agents Oleanolic Acid 6SMK8R7TGJ

Beschreibung
Zusammenfassung:	© 2023 The Authors New Phytologist © 2023 New Phytologist Foundation. Wilforlide A is one of the main active constituents produced in trace amounts in Tripterygium wilfordii Hook F, which has excellent anti-inflammatory and immune suppressive effects. Despite the seeming structural simplicity of the compound, the biosynthetic pathway of wilforlide A remains unknown. Gene-specific expression analysis and genome mining were used to identify the gene candidates, and their functions were studied in vitro and in vivo. A modularized two-step (M2S) technique and CRISPR-Cas9 methods were used to construct engineering yeast. Here, we identified a cytochrome P450, TwCYP82AS1, that catalyses C-22 hydroxylation during wilforlide A biosynthesis. We also found that TwCYP712K1 to K3 can further oxidize the C-29 carboxylation of oleanane-type triterpenes in addition to friedelane-type triterpenes. Reconstitution of the biosynthetic pathway in engineered yeast increased the precursor supply, and combining TwCYP82AS1 and TwCYP712Ks produced abrusgenic acid, which was briefly acidified to achieve the semisynthesis of wilforlide A. Our work presents an alternative metabolic engineering approach for obtaining wilforlide A without relying on extraction from plants
Beschreibung:	Date Completed 26.01.2024 Date Revised 26.01.2024 published: Print-Electronic Citation Status MEDLINE
ISSN:	1469-8137
DOI:	10.1111/nph.19427