Frontiers of Chemical Science and Engineering >
Assembly of biosynthetic pathways in Saccharomyces cerevisiae using a marker recyclable integrative plasmid toolbox
Received date: 21 May 2016
Accepted date: 09 Aug 2016
Published date: 17 Mar 2017
Copyright
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) developed in our research group, which has features of ready-to-use, convenient marker recycling, arbitrary element replacement, shuttle plasmid, auxotrophic marker independence, 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.
Key words： pathway assembly; toolbox; reiterative recombination; S. cerevisiae; biosynthesis
Lidan Ye , Xiaomei Lv , Hongwei Yu . Assembly of biosynthetic pathways in Saccharomyces cerevisiae using a marker recyclable integrative plasmid toolbox[J]. Frontiers of Chemical Science and Engineering, 2017 , 11(1) : 126 -132 . DOI: 10.1007/s11705-016-1597-8
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