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 mark...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.展开更多
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.
文摘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.
