Improving the homologous recombination efficiency of Yarrowia lipolytica by grafting heterologous component from Saccharomyces cerevisiae
File(s)
Published version
Author(s)
Type
Journal Article
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 end-joining 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.
Date Issued
2020-12-01
Date Acceptance
2020-11-09
Citation
Metabolic Engineering Communications, 2020, 11
ISSN
2214-0301
Publisher
Elsevier
Journal / Book Title
Metabolic Engineering Communications
Volume
11
Copyright Statement
©2020 The Authors. Published by Elsevier B.V. on behalf of International Metabolic Engineering Society. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Sponsor
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/33294367
PII: S2214-0301(20)30052-3
Grant Number
BB/R01602X/1
Subjects
Homologous recombination
Non-homologous end-joining
Oleaginous yeast
Rad52
Yarrowia lipolytica
Publication Status
Published
Coverage Spatial
Netherlands
Article Number
ARTN e00152
Date Publish Online
2020-11-13