Engineering the microenvironment of P450s to enhance the production of diterpenoids in Saccharomyces cerevisiae  

作　　者：Yatian Cheng Linglong Luo Hao Tang Jian Wang Li Ren Guanghong Cui Yujun Zhao Jinfu Tang Ping Su Yanan Wang Yating Hu Ying Ma Juan Guo Luqi Huang 

机构地区：[1]National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Science,State Key Laboratory of Dao-di Herbs,Beijing 100700,China [2]School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China [3]School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China

出　　处：《Acta Pharmaceutica Sinica B》2024年第10期4608-4618,共11页药学学报（英文版）

基　　金：supported by the National Key R&D Program of China(2020YFA0908000);Scientific and technological innovation project of China Academy of Chinese Medical Sciences(CI2023D002,CI2023E002,CI2021A04110,CI2021B014);the National Natural Science Foundation of China(81822046);Key project at central government level:The ability to establish sustainable use of valuable Chinese medicine resources(2060302).

摘　　要：Cytochrome P450 enzymes play a crucial role as catalysts in the biosynthesis of numerous plant natural products(PNPs).Enhancing the catalytic activity of P450s in host microorganisms is essential for the efficient production of PNPs through synthetic biology.In this study,we engineered Saccharomyces cerevisiae to optimize the microenvironment for boosting the activities of P450s,including coexpression with the redox partner genes,enhancing NADPH supply,expanding the endoplasmic reticulum(ER),strengthening heme biosynthesis,and regulating iron uptake.This created a platform for the efficient production 11,20-dihydroxyferruginol,a key intermediate of the bioactive compound tanshinones.The yield was enhanced by 42.1-fold through 24 effective genetic edits.The optimized strain produced up to 67.69±1.33 mg/L 11,20-dihydroxyferruginol in shake flasks.Our work represents a promising advancement toward constructing yeast cell factories containing P450s and paves the way for microbial biosynthesis of tanshinones in the future.

关 键 词：Saccharomyces cerevisiae Metabolic engineering P450s TANSHINONES 11 20-Dihydroxyferruginol 

分 类 号：R915[医药卫生—微生物与生化药学]