Overexpression of Citrate Synthase Increases Isocitric Acid Biosynthesis in the Yeast Yarrowia lipolytica

Piotr Hapeta, Magdalena Rakicka-Pustułka, Piotr Juszczyk, Małgorzata Robak, Waldemar Rymowicz and Zbigniew Lazar
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Piotr Hapeta: Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland
Magdalena Rakicka-Pustułka: Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland
Piotr Juszczyk: Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland
Małgorzata Robak: Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland
Waldemar Rymowicz: Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland
Zbigniew Lazar: Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland

Sustainability, 2020, vol. 12, issue 18, 1-15

Abstract: Yarrowia lipolytica is a non-conventional yeast producing valuable compounds, such as citric acids, from renewable raw materials. This study investigated the impact of citrate synthase overexpression on the biosynthesis of citric and isocitric acid in Y. lipolytica. Two transformants of Y. lipolytica A101.1.31 strain (efficient citric acid producer), overexpressing CIT1 or CIT2 gene (encoding proteins with citrate synthase activity), were constructed. The results revealed that overexpression of either of these genes enhances citrate synthase activity. Additionally, the cit1 knockout strain was unable to use propionate as the sole carbon source, which proves that CIT1 gene encodes a dual activity protein–citrate and 2-methylcitrate synthase. In the overexpressing mutants, a significant increase in isocitric acid biosynthesis was observed. Both CIT1 and CIT2 overexpressing strains produced citric and isocitric acid from vegetable oil in a ratio close to 1 (CA/ICA ratio for wild-type strain was 4.12).

Keywords: Yarrowia lipolytica; citrate synthase; 2-methylcitrate synthase; citrate; isocitrate (search for similar items in EconPapers)
JEL-codes: O13 Q Q0 Q2 Q3 Q5 Q56 (search for similar items in EconPapers)
Date: 2020
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