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Effects of pyruvate dehydrogenase subunits overexpression on the α-ketoglutarate production in Yarrowia lipolytica WSH-Z06

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Abstract

Yarrowia lipolytica WSH-Z06 harbours a promising capability to oversynthesize α-ketoglutarate (α-KG). Its wide utilization is hampered by the formation of high concentrations of pyruvate. In this study, a metabolic strategy for the overexpression of the α and β subunits of pyruvate dehydrogenase E1, E2 and E3 components was designed to reduce the accumulation of pyruvate. Elevated expression level of α subunit of E1 component improved the α-KG production and reduced the pyruvate accumulation. Due to a reduction in the acetyl-CoA supply, neither the growth of cells nor the synthesis of α-KG was restrained by the overexpression of β subunit of E1, E2 and E3 components. Furthermore, via the overexpression of these thiamine pyrophosphate (TPP)-binding subunits, the dependency of pyruvate dehydrogenase on thiamine was diminished in strains T1 and T2, in which α and β subunits of E1 component were separately overexpressed. In these two recombinant strains, the accumulation of pyruvate was insensitive to variations in exogenous thiamine. The results suggest that α-KG production can be enhanced by altering the dependence on TPP of pyruvate dehydrogenase and that the competition for the cofactor can be switched to ketoglutarate dehydrogenase via separate overexpression of the TPP-binding subunits of pyruvate dehydrogenase. The results presented here provided new clue to improve α-KG production.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31130043, 21276109), the Natural Science Foundation of Jiangsu Province (BK2011004), the Author of National Excellent Doctoral Dissertation of PR China (FANEDD, 2011046), the Program for New Century Excellent Talents in the University (NCET-12-0876), the Fundamental Research Funds for the Central Universities (JUSRP51307A) 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

    Hongwei Guo, Guocheng Du, Jingwen Zhou & Jian Chen

  2. Synergetic Innovation Center of Food Safety and Nutrition, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China

    Hongwei Guo, Guocheng Du, 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. Hongwei Guo
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  2. Catherine Madzak
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  3. Guocheng Du
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  4. Jingwen Zhou
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  5. Jian Chen
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Correspondence to Jingwen Zhou or Jian Chen.

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Guo, H., Madzak, C., Du, G. et al. Effects of pyruvate dehydrogenase subunits overexpression on the α-ketoglutarate production in Yarrowia lipolytica WSH-Z06. Appl Microbiol Biotechnol 98, 7003–7012 (2014). https://doi.org/10.1007/s00253-014-5745-0

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  • DOI: https://doi.org/10.1007/s00253-014-5745-0

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