Assembly of biosynthetic pathways in Saccharomyces cerevisiae using a marker recyclable integrative plasmid toolbox  

作　　者：Lidan Ye Xiaomei Lv Hongwei Yu 

机构地区：[1]Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China [2]Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University, Hangzhou 310027, China

出　　处：《Frontiers of Chemical Science and Engineering》2017年第1期126-132,共7页化学科学与工程前沿（英文版）

基　　金：This work was financially supported by the National Natural Science Foundation of China （Grant Nos. 21406196 and 21576234）, Zhejiang Provincial Natural Science Foundation of China （Grant No. LQI4B060005）, and Qianiiang Talents Project.

摘　　要：A robust and versatile tool for multigene pathway assembly is a key to the biosynthesis of high- value chemicals. Here we report the rapid construction of biosynthetic pathways in Saccharomyces cerevisiae using a marker recyclable integrative toolbox （pUMRI） devel- oped in our research group, which has features of ready-to- use, convenient marker recycling, arbitrary element replacement, shuttle plasmid, auxotrophic marker inde- pendence, GAL regulation, and decentralized assembly. Functional isoprenoid biosynthesis pathways containing 4-11 genes with lengths ranging from -10 to -22 kb were assembled using this toolbox within 1-5 rounds of reiterative recombination. In combination with GAL- regulated metabolic engineering, high production of isoprenoids （e.g., 16.3 mg.g-1 dcw carotenoids）was achieved. These results demonstrate the wide range of application and the efficiency of the pUMRI toolbox in multigene pathway construction of S. cerevisiae.A robust and versatile tool for multigene pathway assembly is a key to the biosynthesis of high- value chemicals. Here we report the rapid construction of biosynthetic pathways in Saccharomyces cerevisiae using a marker recyclable integrative toolbox （pUMRI） devel- oped in our research group, which has features of ready-to- use, convenient marker recycling, arbitrary element replacement, shuttle plasmid, auxotrophic marker inde- pendence, GAL regulation, and decentralized assembly. Functional isoprenoid biosynthesis pathways containing 4-11 genes with lengths ranging from -10 to -22 kb were assembled using this toolbox within 1-5 rounds of reiterative recombination. In combination with GAL- regulated metabolic engineering, high production of isoprenoids （e.g., 16.3 mg.g-1 dcw carotenoids）was achieved. These results demonstrate the wide range of application and the efficiency of the pUMRI toolbox in multigene pathway construction of S. cerevisiae.

关 键 词：pathway assembly TOOLBOX reiterative recom-bination S. cerevisiae biosynthesis 

分 类 号：Q946.88[生物学—植物学] TS261.11[轻工技术与工程—发酵工程]