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Redirection of the NADH oxidation pathway in Torulopsis glabrata leads to an enhanced pyruvate production

  • Applied Microbial and Cell Physiology
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Abstract

This study aimed at increasing the pyruvate productivity of a multi-vitamin auxotrophic yeast Torulopsis glabrata by redirecting NADH oxidation from adenosine triphosphate (ATP)-production pathway (oxidative phosphorylation pathway) to non-ATP production pathway (fermentative pathway). Two respiratory-deficient mutants, RD-17 and RD-18, were screened and selected after ethidium bromide (EtBr) mutagenesis of the parent strain T. glabrata CCTCC M202019. Compared with the parent strain, cytochrome aa 3 and b in electron transfer chain (ETC) of RD-18 and cytochrome b in RD-17 were disrupted. As a consequence, the activities of key ETC enzymes of the mutant RD-18, including F0F1-ATP synthase, complex I, complex I + III, complex II + III, and complex IV, decreased by 22.2, 41.6, 53.1, 23.6, and 84.7%, respectively. With the deficiency of cytochromes in ETC, a large amount of excessive cytosolic NADH was accumulated, which hampered the further increase of the glycolytic flux. An exogenous electron acceptor, acetaldehyde, was added to the strain RD-18 culture to oxidize the excessive NADH. Compared with the parent strain, the concentration of pyruvate and the glucose consumption rate of strain RD-18 were increased by 26.5 and 17.6%, respectively, upon addition of 2.1 mM of acetaldehyde. The strategy for increasing the glycolytic flux in T. glabrata by redirecting the NADH oxidation pathway may provide an alternative approach to enhance the glycolytic flux in yeast.

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Acknowledgements

The authors thank the financial support by the Natural Science Foundation of Jiangsu Province of China (contract No. BK2002072); the Scientific Research Foundation for the Returned Overseas Chinese Scholars, Ministry of Education of China; the Ph.D. Programs Foundation of Ministry of Education of China (No. 20040294003); and the Post-graduate Innovation Program of Jiangsu Province, China.

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  1. School of Biotechnology, Southern Yangtze University, 170 , Huihe Road, Wuxi, 214036, People’s Republic of China

    Liming Liu, Yin Li, Guocheng Du & Jian Chen

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  1. Liming Liu
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  2. Yin Li
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  3. Guocheng Du
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  4. Jian Chen
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Correspondence to Jian Chen.

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Liu, L., Li, Y., Du, G. et al. Redirection of the NADH oxidation pathway in Torulopsis glabrata leads to an enhanced pyruvate production. Appl Microbiol Biotechnol 72, 377–385 (2006). https://doi.org/10.1007/s00253-005-0284-3

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  • DOI: https://doi.org/10.1007/s00253-005-0284-3

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