Modification of C20 oxidase in tanshinone biosynthesis pathway

• Published in: Zhongguo zhongyao zazhi Vol. 48; no. 9; p. 2298
• Main Authors: Cao, Xiao-Qing, Ma, Xiao-Hui, Cheng, Ya-Tian, Li, Qi-Shuang, Bu, Jun-Ling, Ma, Ying, Guo, Juan
• Format: Journal Article
• Language: Chinese
• Published: China 01.05.2023
• Subjects: 11-hydroxysugiol; continuous oxidation; cytochrome P450; mutants; directed evolution
• ISSN: 1001-5302
• DOI: 10.19540/j.cnki.cjcmm.20230215.102

Tanshinones are one of the main effective components of Salvia miltiorrhiza, which play important roles in the treatment of cardiovascular diseases. Microbial heterogony production of tanshinones can provide a large number of raw materials for the production of traditional Chinese medicine(TCM) preparations containing S. miltiorrhiza, reduce the extraction cost, and relieve the pressure of clinical medication. The biosynthetic pathway of tanshinones contains multiple P450 enzymes, and the catalytic element with high efficiency is the basis of microbial production of tanshinones. In this study, the protein modification of CYP76AK1, a key P450-C20 hydroxylase in tanshinone pathway, was researched. The protein modeling methods SWISS-MODEL, Robetta, and AlphaFold2 were used, and the protein model was analyzed to obtain the reliable protein structure. The semi-rational design of mutant protein was carried out by molecular docking and homologous alignment. The key amino acid sites affecting the oxidation activity o