文献类型：期刊文献

英文题名：Yeast transcriptional device libraries enable precise synthesis of value-added chemicals from methanol

作者：Zhu, Qiaoyun[1];Liu, Qi[1];Yao, Chaoying[1];Zhang, Yuanxing[1,2];Cai, Menghao[1,3]

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;[3]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Me, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2022

卷号：50

期号：17

起止页码：10187

外文期刊名：NUCLEIC ACIDS RESEARCH

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000852783100001)】；

基金：This study was supported by the National Key R&D Program of China [2020YFA0907800]; National Natural Science Foundation of China [31870073 and 42176238]; Shanghai Rising-Star Program [19QA1402700]; Shanghai Frontiers Science Center ofOptogeneticTechniques for Cell Metabolism [ShanghaiMunicipal Education Commission]; and FundamentalResearch Funds for the Shanghai Science and Technology Innovation Action Plan [17JC1402400]. Funding for open access charge: National Key R&D Program of China [2020YFA0907800]; National Natural Science Foundation of China [31870073 and 42176238].

语种：英文

摘要：Natural methylotrophs are attractive methanol utilization hosts, but lack flexible expression tools. In this study, we developed yeast transcriptional device libraries for precise synthesis of value-added chemicals from methanol. We synthesized transcriptional devices by fusing bacterial DNA-binding proteins (DBPs) with yeast transactivation domains, and linking bacterial binding sequences (BSs) with the yeast core promoter. Three DBP-BS pairs showed good activity when working with transactivation domains and the core promoter of P-AOX1 in the methylotrophic yeast, Pichia pastoris. Fine-tuning of the tandem BSs, spacers and differentiated input promoters further enabled a constitutive transcriptional device library (cTRDL) composed of 126 transcriptional devices with an expression strength of 16-520% and an inducible TRDL (iTRDL) composed of 162 methanol-inducible transcriptional devices with an expression strength of 30-500%, compared with P-AOX1. Selected devices from iTRDL were adapted to the dihydromonacolin L biosynthetic pathway by orthogonal experimental design, reaching 5.5-fold the production from the P-AOX1-driven pathway. The full factorial design of the selected devices from the cTRDL was adapted to the downstream pathway of dihydromonacolin L to monacolin J. Monacolin J production from methanol reached 3.0-fold the production from the P-AOX1-driven pathway. Our engineered toolsets ensured multilevel pathway control of chemical synthesis in methylotrophic yeasts.