Four Cytochrome P450 Enzymes Mediate Oxidation Cascades in the Biosynthesis of Cephalotane‐Type Diterpenoids
Abstract
Abstract Cephalotane‐type diterpenoids, a class of natural products exclusively found in Cephalotaxus plants, are well known for their attractive structures and potent biological activities. However, their low natural abundance and intricate cage‐like structures hinder their accessibility. Recently, the identification of a cephalotene synthase ( Cs CTS) has addressed the first committed step in the biosynthesis. However, the enzymes involved in the complex post‐modification of the cephalotene core into structurally diverse cephalotane‐type diterpenoids remain obscure. In this study, we functionally characterised four novel cytochrome P450 enzymes from C. sinensis . These enzymes demonstrate multiple oxidative functions and cooperatively catalyse a cascade of oxidation reactions, including the formation of signature 13,17‐lactone, 5,19‐lactone, and tropone. We further co‐expressed the characterised CYP450 enzymes in combination with Cs CTS to produce a variety of cephalotane‐type diterpenoids, including hainanolidol ( 2 ), mannolide C ( 3 ), mannolide A ( 4 ), and cephinoid H ( 5 ), in Nicotiana benthamiana . Subsequently, harringtonolide ( 1 ) was chemically converted from hainanolidol (3.9 µmol with 10 equiv of Pb(OAc) 4 ) in 87.5% yield. In this study, the biosynthetic pathways of representative cephalotane‐type diterpenoids were elucidated and reconstructed, thereby establishing a foundation for their sustainable production through biosynthesis and/or chemo‐biosynthesis, highlighting the remarkable efficiency of merely five Cephalotaxus ‐specific enzymes in assembling such structurally complex natural products.
Article Details
Authors (13)
Aobo Guo
Jimei Liu
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Changkang Li
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Shuai Wang
Yaotian Han
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Songyang Sui
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Yuxin Wang
Department of Chemistry
Xinxin Yin
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Beilei Yu
State Key Laboratory of Bioactive Substance and Function of Natural Medicines NHC Key Laboratory of Biosynthesis of Natural Products CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Dawei Chen
State Key Laboratory of Chemo and Biosensing, College of Chemistry and Chemical Engineering, Advanced Catalytic Engineering Research Center of the Ministry of Education
Kebo Xie
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China
Ridao Chen
Division of Chemical Biology & Medicinal Chemistry, College of Pharmacy
Jungui Dai
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, NHC Key Laboratory of Biosynthesis of Natural Products, and Beijing Key Laboratory of Non‐Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China