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Enhanced alpha-ketoglutaric acid production in Yarrowia lipolytica WSH-Z06 by regulation of the pyruvate carboxylation pathway

  • Applied genetics and molecular biotechnology
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Abstract

In previous research, a thiamine-auxotrophic yeast for alpha-ketoglutaric acid (KGA) overproduction was screened in our laboratory and named Yarrowia lipolytica WSH-Z06 (CCTCC no. M207143). However, the high concentration of by-products (mainly pyruvate) limited its application on an industrial scale. To enhance KGA production and reduce pyruvate (PA) accumulation, the pyruvate carboxylation pathway was regulated. By overexpressing the pyruvate carboxylase genes ScPYC1 from Saccharomyces cerevisiae and RoPYC2 from Rhizopus oryzae in Y. lipolytica WSH-Z06, the yields of KGA in Y. lipolytica-ScPYC1 and Y. lipolytica-RoPYC2 increased by 24.5 and 35.3 %, and the yields of PA decreased by 51.9 and 69.8 % in shake flasks, respectively. These changes in the expression levels and activities of key intracellular enzymes showed that enhancing the pyruvate carboxylation pathway had successfully redistributed the carbon flux from PA to KGA. Finally, by controlling the pH in a 3-L fermenter, the maximum concentration of KGA in Y. lipolytica-RoPYC2 reached 62.5 g L−1 with an evident decrease in PA yield from 35.2 to 13.5 g L−1.

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Acknowledgments

This work was supported by the Major State Basic Research Development Program of China (973 Program, 2012CB7208022012CB720806), the Key Program of National Natural Science Foundation of China (31130043), the National Natural Science Foundation of China (31171638), the Research Program of Sate Key Laboratory of Food Science and Technology, Jiangnan University (No. SKLF-TS-200901), the Priority Academic Program Development of Jiangsu Higher Education Institutions, and the 111 Project (111-2-06).

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Authors and Affiliations

  1. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China

    Xiaoxia Yin, Guocheng Du, Jingwen Zhou & Jian Chen

  2. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China

    Jingwen Zhou & Jian Chen

  3. UMR1238 Microbiologie et Génétique Moléculaire, INRA/CNRS/AgroPan’s Tech, CBAI, BP 01, 78850, Thiverval-Grignon, France

    Catherine Madzak

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  1. Xiaoxia Yin
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  2. Catherine Madzak
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  3. Guocheng Du
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  5. Jian Chen
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Correspondence to Jingwen Zhou or Jian Chen.

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Yin, X., Madzak, C., Du, G. et al. Enhanced alpha-ketoglutaric acid production in Yarrowia lipolytica WSH-Z06 by regulation of the pyruvate carboxylation pathway. Appl Microbiol Biotechnol 96, 1527–1537 (2012). https://doi.org/10.1007/s00253-012-4192-z

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