Premium
Identification of a Mitochondrial Oxodicarboxylate Carrier in the Oleaginous Yeast Yarrowia lipolytica
Author(s) -
Trotter Pamela J.,
Juco Karen,
McGreer Samantha,
Orsi Natalie,
Pierson Paige N.,
Savage Nowlan,
Tamayo Lizeth,
Nicaud JeanMarc
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.861.4
Subject(s) - yarrowia , saccharomyces cerevisiae , gene , biology , biochemistry , mutant , yeast , homologous recombination , plasmid
In Saccharomyces cerevisiae, the mitochondrial oxodicarboxylate carriers, encoded by the scODC1 and scODC2 genes, are necessary for effective glutamate biosynthesis and fatty acid utilization. Yarrowia lipolytica , an oleaginous yeast, is able to efficiently degrade as well as accumulate large amounts of lipid substrates. The genome of Y. lipolytica encodes one homolog, the YALI0D02629 gene (denoted ylODC1/2 ), which displays 60–64% identity to the scODC1 and scODC2 genes. The ylODC1/2 coding sequence was amplified from genomic DNA by polymerase chain reaction and cloned into a S. cerevisiae expression vector under control of the Gal1 promoter. When transformed with the ylODC1/2 ‐containing plasmid, the inability of the odc1Δodc2Δ mutant of S. cerevisiae to grow on fatty acid medium is corrected, indicating that the Y. lipolytica protein possesses a similar function. The gene was disrupted in Y. lipolytica by insertion of the ylURA3 gene via homologous recombination in a strain lacking the heterologous recombination capability normally found in this organism. Initial growth analysis has revealed no fatty acid growth impairment in the new mutant, suggesting the presence of redundant transport functions, as was observed in S. cerevisiae . Examination of the effects of the ylODC1/2 gene disruption on glutamate and fatty acid metabolism will allow further assessment of its cellular function in Y. lipolytica .