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The isocitrate dehydrogenase gene of oleaginous yeast Lipomyces starkeyi is linked to lipid accumulation

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The oleaginous yeast Lipomyces starkeyi can accumulate intracellular lipids to over 60% of its cell dry mass under a nitrogen-limited condition. We showed that extracellular and intracellular citrate concentrations of L. starkeyi AS 2.1560 increased and the nicotinamide adenine dinucleotide - isocitrate dehydrogenase (NAD+-IDH) activity decreased at the beginning of the lipid accumulation, suggesting that the attenuation of the NAD+-IDH activity might initiate lipid storage. We next cloned the IDH gene by the methods of degenerate PCR and rapid amplification of cDNA ends. Phylogenetic analyses of the evolutionary relationships among LsIDH1, LsIDH2, and other yeast NAD+-IDHs revealed that the L. starkeyi IDH had a closer relationship with the IDHs of Yarrowia lipolytica. Further real-time PCR analysis showed that the expression levels of both LsIDH1 and LsIDH2 decreased concurrently with the evolution of cellular lipids. Our data should be valuable for understanding the biology of oleaginous yeasts and for further strain engineering of L. starkeyi.

La levure oléagineuse Lipomyces starkeyi peut accumuler des lipides intracellulaires à plus de 60% de son poids sec en condition limitante en azote. Nous avons montré que la concentration intracellulaire et extracellulaire de citrate de L. starkeyi AS 2.1560 augmentait et que l’activité de la NAD+ - isocitrate déshydrogénase (NAD+-IDH) diminuait au début de l’accumulation de lipides, suggérant que l’atténuation de l’activité de la NAD+-IDH pourrait amorcer l’entreposage de lipides. Nous avons ensuite cloné le gène IDH par une réaction en chaîne par polymérase (PCR) dégénérée et amplification rapide des extrémités des ADNc. Les analyses phylogéniques de la relation évolutive entre LsIDH1, LsIDH2 et d’autres NAD+-IDH de levure ont révélé que l’IDH de L. starkeyi est plus étroitement reliée à celles de Yarrowia lipolytica. Des analyses par PCR additionnelles ont montré que les niveaux d’expression de LsIDH1 et LsIDH2 diminuent parallèlement avec l’évolution des lipides cellulaires. Nos résultats devraient être utiles pour comprendre la biologie des levures oléagineuses et pour l’ingénierie subséquente de la souche de L. starkeyi.

Document Type: Research Article

Publication date: September 1, 2009

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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