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Improving the homologous recombination efficiency of Yarrowia lipolytica by grafting the heterogenous component from Saccharomyces cerevisiae
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  • Qingchun Ji,
  • Jie Mai,
  • Ying Ding,
  • Yongjun Wei,
  • Rodrigo Ledesma-Amaro,
  • Xiao-Jun Ji
Qingchun Ji
Nanjing Tech University
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Jie Mai
Nanjing Tech University
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Ying Ding
Nanjing Tech University
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Yongjun Wei
Zhengzhou University
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Rodrigo Ledesma-Amaro
Imperial College London
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Xiao-Jun Ji
Nanjing Tech University
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Abstract

The oleaginous non-conventional yeast Yarrowia lipolytica has enormous potential as a microbial platform for the synthesis of various bioproducts. However, while the model yeast Saccharomyces cerevisiae has very high homologous recombination (HR) efficiency, non-homologous recombination is dominant in Y. lipolytica, and foreign genes are randomly inserted into the genome. Consequently, the low HR efficiency greatly restricts the genetic engineering of this yeast. In this study, RAD52, the key component of the HR machinery in S. cerevisiae, was grafted into Y. lipolytica to improve HR efficiency. The gene ade2, whose deletion can result in a brown colony phenotype, was used as the reporter gene for evaluating the HR efficiency. The HR efficiency of Y. lipolytica strains before and after integrating the ScRad52 gene was compared using insets with homology arms of different length. The results showed that the strategy could achieve gene targeting efficiencies of up to 95% with a homology arm length of 1000 bp, which was 6.5 times of the wildtype strain and 1.6 times of the traditionally used ku70 disruption strategy. This study will facilitate the further genetic engineering of Y. lipolytica to make it a more efficient cell factory for the production of value-added compounds.

Peer review status:UNDER REVIEW

29 Aug 2020Submitted to Biotechnology Journal
01 Sep 2020Assigned to Editor
01 Sep 2020Submission Checks Completed
07 Sep 2020Reviewer(s) Assigned