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Regulatory properties of malic enzyme in the oleaginous yeast, Yarrowia lipolytica, and its non-involvement in lipid accumulation

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Abstract

Malic enzyme (EC 1.1.1.40) converts l-malate to pyruvate and CO2 providing NADPH for metabolism especially for lipid biosynthesis in oleaginous microorganisms. However, its role in the oleaginous yeast, Yarrowia lipolytica, is unclear. We have cloned the malic enzyme gene (YALI0E18634g) from Y. lipolytica into pET28a, expressed it in Escherichia coli and purified the recombinant protein (YlME). YlME used NAD+ as the primary cofactor. Km values for NAD+ and NADP+ were 0.63 and 3.9 mM, respectively. Citrate, isocitrate and α-ketoglutaric acid (>5 mM) were inhibitory while succinate (5–15 mM) increased NADP+- but not NAD+-dependent activity. To determine if fatty acid biosynthesis could be increased in Y. lipolytica by providing additional NADPH from an NADP+-dependent malic enzyme, the malic enzyme gene (mce2) from an oleaginous fungus, Mortierella alpina, was expressed in Y. lipolytica. No significant changes occurred in lipid content or fatty acid profiles suggesting that malic enzyme is not the main source of NADPH for lipid accumulation in Y. lipolytica.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31271812,81071685,21276108), the National High Technology Research and Development Program of China (863 Program 2012AA022105C), the National Basic Research Program of China 973 Program (2012CB720802), the National Science Fund for Distinguished Young Scholars (31125021), the 111 project B07029, Starting Grant from Institut Mérieux (Strategic Mérieux Research Grants), and Institut Mérieux Fellowship 2012.

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Authors and Affiliations

  1. School of Food Science and Technology, Jiangnan University, Wuxi, 214122, Jiangsu, People’s Republic of China

    Huaiyuan Zhang, Luning Zhang, Haiqin Chen, Yong Q. Chen, Colin Ratledge, Yuanda Song & Wei Chen

  2. Department of Biological Sciences, University of Hull, Hull, HU6 7RX, UK

    Colin Ratledge

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  1. Huaiyuan Zhang
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  2. Luning Zhang
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  3. Haiqin Chen
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  4. Yong Q. Chen
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  5. Colin Ratledge
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  6. Yuanda Song
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  7. Wei Chen
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Correspondence to Yuanda Song.

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Zhang, H., Zhang, L., Chen, H. et al. Regulatory properties of malic enzyme in the oleaginous yeast, Yarrowia lipolytica, and its non-involvement in lipid accumulation. Biotechnol Lett 35, 2091–2098 (2013). https://doi.org/10.1007/s10529-013-1302-7

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  • DOI: https://doi.org/10.1007/s10529-013-1302-7

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