NDLI: Identification of a Yarrowia lipolytica acetamidase and its use as a yeast genetic marker

Content Provider	Springer Nature : BioMed Central
Author	Hamilton, Maureen Consiglio, Andrew L. MacEwen, Kyle Shaw, A. Joe Tsakraklides, Vasiliki
Abstract	Background Yarrowia lipolytica is an oleaginous yeast that can be genetically engineered to produce lipid and non-lipid biochemicals from a variety of feedstocks. Metabolic engineering of this organism usually requires genetic markers in order to select for modified cells. The potential to combine multiple genetic manipulations depends on the availability of multiple or recyclable selectable markers. Results We found that Y. lipolytica has the ability to utilize acetamide as the sole nitrogen source suggesting that the genome contains an acetamidase gene. Two potential Y. lipolytica acetamidase gene candidates were identified by homology to the A. nidulans acetamidase amdS. These genes were deleted in the wild-type Y. lipolytica strain YB-392, and deletion strains were evaluated for acetamide utilization. One deletion strain was unable to grow on acetamide and a putative acetamidase gene YlAMD1 was identified. Transformation of YlAMD1 followed by selection on acetamide media and counterselection on fluoroacetamide media showed that YlAMD1 can be used as a recyclable genetic marker in Saccharomyces cerevisiae and Ylamd1Δ Y. lipolytica. Conclusions These findings add to our understanding of Y. lipolytica nitrogen utilization and expand the set of genetic tools available for engineering this organism, as well as S. cerevisiae.
Related Links	https://microbialcellfactories.biomedcentral.com/counter/pdf/10.1186/s12934-020-1292-9.pdf
Ending Page	9
Page Count	9
Starting Page	1
File Format	HTM / HTML
ISSN	14752859
DOI	10.1186/s12934-020-1292-9
Journal	Microbial Cell Factories
Issue Number	1
Volume Number	19
Language	English
Publisher	BioMed Central
Publisher Date	2020-02-05
Access Restriction	Open
Subject Keyword	Applied Microbiology Biotechnology Microbiology Microbial Genetics and Genomics Enzymology Genetic Engineering Yarrowia lipolytica Acetamidase Acetamide Fluoroacetamide Dominant marker Counterselectable marker Marker removal Marker recycling
Content Type	Text
Resource Type	Article
Subject	Applied Microbiology and Biotechnology Bioengineering Biotechnology
Journal Impact Factor	4.3/2023
5-Year Journal Impact Factor	5.5/2023

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