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

Author:

Liu Yuan12ORCID,Zhou Jiawei13ORCID,Liu Panting1,Hu Tianyuan4,Liu Xuan1,Gao Jie1,Ma Lin1,Lu Yun1ORCID,Li Dan5,Jiang Zhouqian1ORCID,Zhang Xianan1,Huang Luqi2,Gao Wei1ORCID,Wu Xiaoyi1ORCID,Zhang Yifeng2ORCID,Liu Changli1ORCID

Affiliation:

1. School of Traditional Chinese Medicine Capital Medical University Beijing 100069 China

2. National Key Laboratory for Quality Ensurance and Sustainable Use of Dao‐di Herbs, National Resource Center for Chinese Materia Medica China Academy of Chinese Medical Sciences Beijing 100700 China

3. College of Biotechnology and Bioengineering Zhejiang University of Technology Hangzhou 310014 China

4. School of Pharmacy, College of Medicine Hangzhou Normal University Hangzhou 311121 China

5. School of Pharmaceutical Science Capital Medical University Beijing 100069 China

Abstract

Summary 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.

Publisher

Wiley

Subject

Plant Science,Physiology

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3