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Improved production of ethanol by deleting FPS1 and over-expressing GLT1 in Saccharomyces cerevisiae

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Abstract

To improve ethanol production in Saccharomyces cerevisiae, two yeast strains were constructed. In the mutant KAM-3, the FPS1 gene, which encodes a channel protein responsible for glycerol export, was deleted. The mutant KAM-11 had the GLT1 gene (encoding glutamate synthase) placed under the PGK1 promoter while having the FPS1 deletion. Growth rate and biomass concentration remained virtually unchanged with the mutant KAM-11, compared to that of the parent. Over-expression of GLT1 by the PGK1 promoter along with FPS1 deletion resulted in a 14% higher ethanol production and a 30% lower glycerol formation compared to the parental strain under anaerobic fermentation conditions. Furthermore, acetate and pyruvic acid formation was also reduced in order for cells to maintain redox balance.

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References

  • Bakker BM, Overkamp KM, Van Maris AJ, Kötter P, Luttik MA, van Dijken JP, Pronk JT (2001) Stoichiometry and compartmentation of NADH metabolism in Saccharomyces cerevisiae. FEMS Microbiol Rev 25:15–37

    Article  PubMed  CAS  Google Scholar 

  • Björkqvist S, Ansell R, Adler L, Lidén G (1997) Physiological response to anaerobicity of glycerol-3-phosphate dehydrogenase mutants of Saccharomyces cerevisiae. Appl Environ Microbiol 63:128–132

    PubMed  Google Scholar 

  • Hedfalk K, Bill RM, Jonathan GL, Mullins GL, Karlgren S, Filipsson C, Bergstrom J, Tamás MJ, Rydström J, Hohmann S (2004) A regulatory domain in the C-terminal extension of the yeast glycerol channel Fps1p. J Biol Chem 158:14954–14960

    Article  Google Scholar 

  • Hohmann S (2002) Osmotic stress signaling and osmoadaptation in yeasts. Microbiol Mol Biol Rev 66:300–372

    Article  PubMed  CAS  Google Scholar 

  • Michnick S, Roustan JL, Remize F, Barre P (1997) Modulation of glycerol and ethanol yields during alcoholic fermentation in Saccharomyces cerevisiae strains overexpressed or disrupted for GPD1 encoding glycerol 3-phosphate dehydrogenase. Yeast 13:783–793

    Article  PubMed  CAS  Google Scholar 

  • Nissen TL, Kielland-Brandt MC, Nielsen J, Villadsen J (2000) Optimization of ethanol production in Saccharomyces cerevisiae by metabolic engineering of the ammonium assimilation. Metab Eng 2:69–77

    Article  PubMed  CAS  Google Scholar 

  • Oliveira R, Lages F, Silva-Graça M, Lucas C (2003) Fps1p channel is the mediator of the major part of glycerol passive diffusion in Saccharomyces cerevisiae: artefacts and re-definitions. Biochim Biophys Acta 1613:57–71

    Article  PubMed  CAS  Google Scholar 

  • Oner ET, Oliver SG, Kirdar B (2005) Production of ethanol from starch by respiration-deficient recombinant Saccharomyces cerevisiae. Appl Environ Microbiol 71:6443–6445

    Article  CAS  Google Scholar 

  • Pahlman AK, Granath K, Ansell R, Hohmann S, Adler L (2001) The yeast glycerol 3-phosphatases Gp1p and Gp2p are required for glycerol biosynthesis and differentially involved in the cellular responses to osmotic, anaerobic and oxidative stress. J Biol Chem 276:3555–3563

    Article  PubMed  CAS  Google Scholar 

  • Remize F, Roustan JL, Sablayrolles JM, Barre P, Dequin S (1999) Glycerol overproduction by engineered Saccharomyces cerevisiae wine yeast strains leads to substantial changes in by-product formation and to a stimulation of fermentation rate in stationary phase. Appl Environ Microbiol 65:143–149

    PubMed  CAS  Google Scholar 

  • Remize F, Barnavon L, Dequin S (2001) Glycerol export and glycerol-3-phosphate dehydrogenase, but not glycerol phosphatase, are rate limiting for glycerol production in Saccharomyces cerevisiae. Metab Eng 3:301–312

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Valadi H, Larsson C, Gustafsson L (1998) Improved ethanol production by glycerol 3-phosphate dehydrogenase mutants of Saccharomyces cerevisiae. Appl Microbiol Biotechnol 4:434–439

    Article  Google Scholar 

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Acknowledgement

The authors would like to thank Prof. Pingsheng Ma for his valuable instruction during the experimental work. This work was financially suported by the Chinese National High-Tech R&D Program (2002AA647040).

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Authors and Affiliations

  1. School of Chemical Engineering and Technology, Tianjin University, Tianjin, China

    Qing-Xue Kong, Li-Min Cao, Ai-Li Zhang, Xun Chen & Xue-Ming Zhao

  2. Department of Food Science, Tianjin Agricultural College, Tianjin, China

    Qing-Xue Kong

  3. College of Life Science and Technology, Jinan University, Guangzhou, China

    Ji-Guang Gu

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  1. Qing-Xue Kong
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  2. Ji-Guang Gu
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  3. Li-Min Cao
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  4. Ai-Li Zhang
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  5. Xun Chen
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  6. Xue-Ming Zhao
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Correspondence to Xue-Ming Zhao.

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Kong, QX., Gu, JG., Cao, LM. et al. Improved production of ethanol by deleting FPS1 and over-expressing GLT1 in Saccharomyces cerevisiae . Biotechnol Lett 28, 2033–2038 (2006). https://doi.org/10.1007/s10529-006-9185-5

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  • DOI: https://doi.org/10.1007/s10529-006-9185-5

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