Importance of the methyl-citrate cycle on glycerol metabolism in the yeast Yarrowia lipolytica |
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Authors: | Seraphim Papanikolaou Athanasios Beopoulos Anna Koletti France Thevenieau Apostolis A. Koutinas Jean-Marc Nicaud George Aggelis |
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Affiliation: | 1. Department of Food Science and Human Nutrition, Agricultural University of Athens, 75 Iera Odos, Athens 11855, Greece;2. INRA, UMR1319, Micalis, F-78352, Jouy-en-Josas, France;3. CNRS, Micalis, F-78352, Jouy-en-Josas, France;4. Unit of Microbiology, Department of Biology, Division of Genetics, Cell and Development Biology, University of Patras, Patras 26500, Greece;5. Department of Biological Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia |
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Abstract: | A novel approach to trigger lipid accumulation and/or citrate production in vivo through the inactivation of the 2-methyl-citrate dehydratase in Yarrowia lipolytica was developed. In nitrogen-limited cultures with biodiesel-derived glycerol utilized as substrate, the Δphd1 mutant (JMY1203) produced 57.7 g/L of total citrate, 1.6-fold more than the wild-type strain, with a concomitant glycerol to citrate yield of 0.91 g/g. Storage lipid in cells increased at the early growth stages, suggesting that inactivation of the 2-methyl-citrate dehydratase would mimic nitrogen limitation. Thus, a trial of JMY1203 strain was performed with glycerol under nitrogen-excess conditions. Compared with the equivalent nitrogen-limited culture, significant quantities of lipid (up to ∼31% w/w in dry weight, 1.6-fold higher than the nitrogen-limited experiment) were produced. Also, non-negligible quantities of citric acid (up to ∼26 g/L, though 0.57-fold lower than the nitrogen-limited experiment) were produced, despite remarkable nitrogen presence into the medium, indicating the construction of phenotype that constitutively accumulated lipid and secreted citrate in Y. lipolytica during growth on waste glycerol utilized as substrate. |
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Keywords: | Yarrowia lipolytica Oleaginous yeast Citric acid Crude glycerol Methylcitrate cycle |
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