Characterization and heterologous reconstitution of Taxus biosynthetic enzymes leading to baccatin III

• 成果类型: Article
• 署名作者: Jiang, Bin; Gao, Lei; Wang, Haijun; Sun, Yaping; Zhang, Xiaolin; Ke, Han; Liu, Shengchao; Ma, Pengchen; Liao, Qinggang; Wang, Yue; Wang, Huan; Liu, Yugeng; Du, Ran; Rogge, Torben; Li, Wei; Shang, Yi; Houk, K. N.; Xiong, Xingyao; Xie, Daoxin; Huang, Sanwen; Lei, Xiaoguang; Yan, Jianbin
• 署名单位: Guangdong Laboratory for Lingnan Modern Agriculture; Chinese Academy of Agricultural Sciences; Agriculture Genomes Institute at Shenzhen, CAAS; Peking University; South China Agricultural University; University of California System; University of California Los Angeles; Yunnan Normal University; Tsinghua University; Shenzhen Bay Laboratory
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-9217
• DOI: 10.1126/science.adj3484
• 发表日期: 2024-02-09
• 页码: 622-629
• 关键词: taxol biosynthesis; molecular-cloning; functional expression; taxadiene synthase; cytochrome-p450; cdna; paclitaxel; derivatives; oxetane; CONSTRUCTION

摘要

Paclitaxel is a well known anticancer compound. Its biosynthesis involves the formation of a highly functionalized diterpenoid core skeleton (baccatin III) and the subsequent assembly of a phenylisoserinoyl side chain. Despite intensive investigation for half a century, the complete biosynthetic pathway of baccatin III remains unknown. In this work, we identified a bifunctional cytochrome P450 enzyme [taxane oxetanase 1 (TOT1)] in Taxus mairei that catalyzes an oxidative rearrangement in paclitaxel oxetane formation, which represents a previously unknown enzyme mechanism for oxetane ring formation. We created a screening strategy based on the taxusin biosynthesis pathway and uncovered the enzyme responsible for the taxane oxidation of the C9 position (T9 alpha H1). Finally, we artificially reconstituted a biosynthetic pathway for the production of baccatin III in tobacco.