文献类型：期刊文献

英文题名：De Novo Production of Plant 4′-Deoxyflavones Baicalein and Oroxylin A from Ethanol in Crabtree-Negative Yeast

作者：Qian, Zhilan[1];Yu, Jiahui[1];Chen, Xinjie[1];Kang, Yijia[1];Ren, Yanna[1];Liu, Qi[1];Lu, Jian[1];Zhao, Qing[2,3];Cai, Menghao[1,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Shanghai Chenshan Plant Sci Res Ctr, Shanghai Key Lab Plant Funct Genom & Resources, Shanghai Chenshan Bot Garden, Shanghai 201602, Peoples R China;[3]Chinese Acad Sci, CAS Ctr Excellence Mol Plant Sci, State Key Lab Plant Mol Genet, Shanghai 200032, Peoples R China;[4]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Me, Sch Pharm, Shanghai 200237, Peoples R China

年份：2022

卷号：11

期号：4

起止页码：1600

外文期刊名：ACS SYNTHETIC BIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000791641700020)】；

基金：This work was supported by the National Key Research and Development Program of China [Grant number 2018YFC1706202], the Natural Science Foundation of China [Grant number 31870073], the Shanghai Rising-Star Program [Grant number 19QA1402700], and Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism [Shanghai Municipal Education Commission, Grant 2021 Sci & Tech 03-28]. The authors thank Prof. Yi Yang and Dr. Xianjun Chen, East China University of Science and Technology, for providing the pADH1 (410)-LVAD plasmid.

语种：英文

外文关键词：plant 4 '-deoxyflavones; Scutellaria baicalensis Georgi; Pichia pastoris; transcriptional regulation device; fine-tuning pathway control

摘要：Baicalein and oroxylin A are well-known medicinal 4'-deoxyflavones found mainly in the roots of traditional medicinal plant Scutellaria baicalensis Georgi. However, extraction from plants is time-consuming, environmentally unfriendly, and insufficient Although microbial synthesis of flavonoids has been extensively reported, synthesis of downstream modified 4'-deoxyflavones has not, and their yields are extremely low. Here, we reassembled the S. baicalensis 4'-deoxyflavone biosynthetic pathway in a Crabtree-negative yeast, Pichia pastoris, with activity analysis and combinatorial expression of eight biosynthetic genes, allowing production of 4'-deoxyflavones like baicalein, oroxylin A, wogonin, norwogonin, 6-methoxywogonin, and the novel 6-methoxynorwogonin. De novo baicalein synthesis was then achieved by complete pathway assembly. Toxic intermediates highly impaired the cell production capacity; hence, we alleviated cinnamic acid growth inhibition by culturing the cells at near-neutral pH and using alcoholic carbon sources. To achieve pathway balance and improve baicalein and oroxylin A synthesis, we further divided the pathway into five modules. A series of ethanol-induced and constitutive transcriptional amplification devices were constructed to adapt to the modules. This fine-tuning pathway control considerably reduced byproduct and intermediate accumulation and achieved high-level de novo baicalein (401.9 mg/L with a total increase of 1182-fold, the highest titer reported) and oroxylin A (339.5 mg/L, for the first time) production from ethanol. This study provides new strategies for the microbial synthesis of 4'-deoxyflavones and other flavonoids.