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

Author

Listed:
  • 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)

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).

Suggested Citation

  • Piotr Hapeta & Magdalena Rakicka-Pustułka & Piotr Juszczyk & Małgorzata Robak & Waldemar Rymowicz & Zbigniew Lazar, 2020. "Overexpression of Citrate Synthase Increases Isocitric Acid Biosynthesis in the Yeast Yarrowia lipolytica," Sustainability, MDPI, vol. 12(18), pages 1-15, September.
    • Handle: RePEc:gam:jsusta:v:12:y:2020:i:18:p:7364-:d:410632
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