文献类型：期刊文献

英文题名：Heterologous pathway assembly reveals molecular steps of fungal terreic acid biosynthesis

作者：Kong, Chuixing[1];Huang, Hezhou[1];Xue, Ying[1];Liu, Yiqi[1];Peng, Qiangqiang[1];Liu, Qi[1];Xu, Qin[1];Zhu, Qiaoyun[1];Yin, Ying[1];Zhou, Xiangshan[1];Zhang, Yuanxing[1,2];Cai, Menghao[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2018

卷号：8

外文期刊名：SCIENTIFIC REPORTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000423787500091)】；

基金：This work was supported by Fundamental Research Funds for the Shanghai Science and Technology Innovation Action Plan (17JC1402400) and Talent Program of School of Biotechnology in East China University of Science and Technology. We thank Dr. Faliang An, East China University of Science and Technology, for the assistance in NMR analysis. The authors declare that they have no conflict of interest. We also gratefully acknowledge Prof. Hong Lu, School of Life Sciences, Fudan University, for kindly providing the yeast two-hybrid system.

语种：英文

摘要：Terreic acid is a potential anticancer drug as it inhibits Bruton's tyrosine kinase; however, its biosynthetic molecular steps remain unclear. In this work, the individual reactions of terreic acid biosynthesis were determined by stepwise pathway assembly in a heterologous host, Pichia pastoris, on the basis of previous knockout studies in a native host, Aspergillus terreus. Polyketide synthase AtX was found to catalyze the formation of partially reduced polyketide 6-methylsalicylic acid, followed by 3-methylcatechol synthesis by salicylate 1-monooxygenase AtA-mediated decarboxylative hydroxylation of 6-methylsalicylic acid. Our results show that cytochrome P450 monooxygenase AtE hydroxylates 3-methylcatechol, thus producing the next product, 3-methyl-1,2,4-benzenetriol. A smaller putative cytochrome P450 monooxygenase, AtG, assists with this step. Then, AtD causes epoxidation and hydroxyl oxidation of 3-methyl-1,2,4-benzenetriol and produces a compound terremutin, via which the previously unknown function of AtD was identified as cyclooxygenation. The final step involves an oxidation reaction of a hydroxyl group by a glucose-methanol-choline oxidoreductase, AtC, which leads to the final product: terreic acid. Functions of AtD and AtG were determined for the first time. All the genes were reanalyzed and all intermediates and final products were isolated and identified. Our model fully defines the molecular steps and corrects previous results from the literature.