Identification and characterization of camptothecin tailoring enzymes in Nothapodytes tomentosa

作     者：Yin Chen Jian-Ping Huang Yong-Jiang Wang Meng-Ling Tu Junheng Li Bingyan Xu Guoqing Peng Jing Yang Sheng-Xiong Huang Yin Chen;Jian-Ping Huang;Yong-Jiang Wang;Meng-Ling Tu;Junheng Li;Bingyan Xu;Guoqing Peng;Jing Yang;Sheng-Xiong Huang

作者机构：State Key Laboratory of Phytochemistry and Plant Resources in West China and Yunnan Key Laboratory of Natural Medicinal ChemistryKunming Institute of BotanyChinese Academy of SciencesKunming 650201China University of Chinese Academy of SciencesBeijing 100049China Key Laboratory for Economic Plants and BiotechnologyKunming Institute of BotanyChinese Academy of SciencesKunming 650201China 

出 版 物：《Journal of Integrative Plant Biology》 (植物学报（英文版）)

年 卷 期：2024年第66卷第6期

页      面：1158-1169页

核心收录：

学科分类：0907[农学-林学] 08[工学] 0829[工学-林业工程] 09[农学] 

基　　金：supported by National Natural Science Foundation of China(82225043) Biological Resources Program(KFJBRP-009) Yunnan Revitalization Talent Support Program“Yunling Scholar”Project(S.-X.H.) Yunnan Revitalization Talent Support Program“Young Talent”Project(J.-P.H.) 

主　　题：camptothecin biosynthesis enzymatic assay Nothapodytes tomentosa tailoring enzyme transcriptomic analysis 

摘      要：Camptothecin is a complex monoterpenoid indole alkaloid with remarkable antitumor *** that two C-10 modified camptothecin derivatives,topotecan and irinotecan,have been approved as potent anticancer agents,there is a critical need for methods to access other aromatic ringfunctionalized congeners(e.g.,C-9,C-10,etc.).However,contemporary methods for chemical oxidation are generally harsh and low-yielding when applied to the camptothecin scaffold,thereby limiting the development of modified *** herein,we have identified four tailoring enzymes responsible for C-9 modifications of camptothecin from Nothapodytes tomentosa,via metabolomic and transcriptomic *** consist of a cytochrome P450(Nt CPT9H)which catalyzes the regioselective oxidation of camptothecin to 9-hydroxycamptothecin,as well as two methyltransferases(Nt OMT1/2,converting 9-hydroxycamptothecin to 9-methoxycamptothecin),and a uridine diphosphate-glycosyltransferase(Nt UGT5,decorating 9-hydroxycamptothecin to9-β-D-glucosyloxycamptothecin).Importantly,the critical residues that contribute to the specific catalytic activity of Nt CPT9H have been elucidated through molecular docking and mutagenesis *** work provides a genetic basis for producing camptothecin derivatives through metabolic *** will hasten the discovery of novel C-9 modified camptothecin derivatives,with profound implications for pharmaceutical manufacture.