Frontiers of Chemical Science and Engineering

, Volume 11, Issue 1, pp 126–132 | Cite as

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

Communication
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Abstract

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.

Keywords

pathway assembly toolbox reiterative recombination S. cerevisiae biosynthesis 

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Notes

Acknowledgements

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. LQ14B060005), and Qianjiang Talents Project.

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© Higher Education Press and Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Institute of Bioengineering, College of Chemical and Biological EngineeringZhejiang UniversityHangzhouChina
  2. 2.Key Laboratory of Biomass Chemical Engineering of Ministry of EducationZhejiang UniversityHangzhouChina

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