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Metabolic engineering of Saccharomyces cerevisiae for increased bioconversion of lignocellulose to ethanol

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Abstract

The absence of pentose-utilizing enzymes in Saccharomyces cerevisiae is an obstacle for efficiently converting lignocellulosic materials to ethanol. In the present study, the genes coding xylose reductase (XYL1) and xylitol dehydrogenase (XYL2) from Pichia stipitis were successfully engineered into S. cerevisae. As compared to the control transformant, engineering of XYL1 and XYL2 into yeasts significantly increased the microbial biomass (8.1 vs. 3.4 g/L), xylose consumption rate (0.15 vs. 0.02 g/h) and ethanol yield (6.8 vs. 3.5 g/L) after 72 h fermentation using a xylose-based medium. Interestingly, engineering of XYL1 and XYL2 into yeasts also elevated the ethanol yield from sugarcane bagasse hydrolysate (SUBH). This study not only provides an effective approach to increase the xylose utilization by yeasts, but the results also suggest that production of ethanol by this recombinant yeasts using unconventional nutrient sources, such as components in SUBH deserves further attention in the future.

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References

  1. Watanabe S, Saleh AA, Pack SP, Annaluru N, Kodaki T, Makino K (2007) Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing protein engineered NADP+-dependent xylitol dehydrogenase. J Biotechnol 130:316–319

    Article  PubMed  CAS  Google Scholar 

  2. Palmqvist E, Hahn-Hägerdal B (2000) Fermentation of lignocellulosic hydrolysates. II: inhibitors and mechanisms of inhibition. Bioresour Technol 74:25–33

    Article  CAS  Google Scholar 

  3. Hahn-Hägerdal B, Galbe M, Gorwa-Grauslund G, Liden G, Zacchi G (2006) Bio-ethanol. the fuel of tomorrow from the residues of today. Trends Biotechnol 24:549–556

    Article  PubMed  Google Scholar 

  4. McMillan JD, Boynton BL (1994) Arabinose utilization by xylose-fermenting yeasts and fungi. Appl Biochem Biotehcnol 45–46:569–584

    Article  Google Scholar 

  5. Smiley KL, Bolen PL (1982) Demonstration of d-xylose reductase and d-xylitol dehydrogenase in Pachysolen tannophilus. Biotechnol Lett 4:607–610

    Article  CAS  Google Scholar 

  6. Aristidou A, Penttilä M (2000) Metabolic engineering applications to renewable resource utilization. Curr Opin Biotechnol 11:187–198

    Article  PubMed  CAS  Google Scholar 

  7. Hahn-Hägerdal B, Wahlbom CF, Gardonyi M, Van Zyl WH, Cordero Otero RR, Jönsson LJ (2001) Metabolic engineering of Saccharomyces cerevisiae for xylose utilization. Adv Biochem Eng Biotechnol 73:53–84

    PubMed  Google Scholar 

  8. Hahn-Hägerdal B, Karhumaa K, Jeppsson M, Gorwa-Grauslund MF (2007) Metabolic engineering for pentose utilization in Saccharomyces cerevisiae. Adv Biochem Eng Biotechnol 108:147–177

    PubMed  Google Scholar 

  9. Amore R, Kötter P, Küster C, Ciriacy M, Hollenberg CP (1991) Cloning and expression in Saccharomyces cerevisiae of the NAD(P)H-dependent xylose reductase-encoding gene (XYL1) from the xylose-assimilating yeast Pichia stipitis. Gene 109:89–97

    Article  PubMed  CAS  Google Scholar 

  10. Shi NQ, Prahl K, Hendrick J, Cruz J, Lu P, Cho JY, Jones S, Jeffries T (2000) Characterization and complementation of a Pichia stipitis mutant unable to grow on d-xylose or l-arabinose. Appl Biochem Biotechnol 84–86:201–216

    Article  PubMed  Google Scholar 

  11. Sambrook J, Russel DW (2001) Molecular cloning—a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  12. Watanabe S, Kodaki T, Makino K (2005) Complete reversal of coenzyme specificity of xylitol dehydrogenase and increase of thermostability by the introduction of structural zinc. J Biol Chem 280:10340–10349

    Article  PubMed  CAS  Google Scholar 

  13. Bradford MM (1976) A rapid sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  14. Martin C, Alriksson B, Sjöde A, Nilvebrant NO, Jönsson LJ (2007) Dilute-sulphuric acid prehydrolysis of agricultural and agro-industrial residues for ethanol production. Appl Biochem Biotechnol 137–140:339–352

    Article  PubMed  Google Scholar 

  15. Alriksson B, Rose SH, Van Zyl WH, Sjöde A, Nilvebrant NO, Jönsson LJ (2009) Cellulase production from spent lignocellulose hydrolysates by recombinant Aspergillus niger. Appl Environ Microbiol 75:2366–2374

    Article  PubMed  CAS  Google Scholar 

  16. Lang X, Macdonald DG, Hill GA (2001) Recycle bioreactor for bioethanol production from wheat starchII. Fermentation and economics. Energy Sour 23:427–436

    Article  CAS  Google Scholar 

  17. Demirbas A (2005) Bioethanol from cellulosic materials: a renewable motor fuel from biomass. Energy Sour 27:327–337

    Article  CAS  Google Scholar 

  18. Eliasson A, Christensson C, Wahlbom CF, Hahn-Hägerdal B (2009) Anaerobic xylose fermentation by recombinant Saccharomyces cerevisiae carrying XYL1, XYL2, and XKS1 in mineral medium chemostat cultures. Appl Environ Microbiol 66:3381–3386

    Article  Google Scholar 

  19. Roca C, Nielsen J, Olsson L (2003) Metabolic engineering of ammonium assimilation in xylose fermenting Saccharomyces cerevisiae improves ethanol production. Appl Environ Microbiol 69:4732–4736

    Article  PubMed  CAS  Google Scholar 

  20. Pitkänen JP, Aristidou A, Salusjärvi L, Ruohonen L, Penttilä M (2003) Metabolic flux analysis of xylose metabolism in recombinant Saccharomyces cerevisiae using continuous culture. Metab Eng 5:16–31

    Article  PubMed  Google Scholar 

  21. Kuyper M, Harhangi HR, Stave AK, Winkler AA, Jetten MS, de Laat WT, den Ridder JJ, Op den Camp HJ, van Dijken JP, Pronk JT (2003) High-level functional expression of a fungal xylsoe isomerase: the key to efficient ethanolic fermentation of xylose by Saccharomyces cerevisiae. FEMS Yeast Res 4:69–78

    Article  PubMed  CAS  Google Scholar 

  22. Zaldivar J, Borges A, Johansson B, Smits HP, Villas-Boas SG, Nielsen J, Olsson L (2002) Fermentation performance and intracellular metabolite patterns in laboratory and industrial xylose-fermenting Saccharomyces cerevisiae. Appl Microbiol Biotechnol 59:436–442

    Article  PubMed  CAS  Google Scholar 

  23. Sellick CA, Campbell RN, Reece RJ (2008) Galactose metabolism in yeast-structure and regulation of the leloir pathway enzymes and the genes encoding them. Int Rev Cell Mol Biol 269:111–150

    Article  PubMed  CAS  Google Scholar 

  24. Bruinenberg PM, De Bot PHM, Van Dijken JP, Scheffers TW (1984) NADH-linked aldose reductase: the key to anaerobic alcoholic fermentation of xylose by yeasts. Appl Microbiol Biotechnol 19:256–260

    Article  CAS  Google Scholar 

  25. Bruinenberg PM, De Bot PHM, Van Dijken JP, Scheffers WA (1983) The role of redox balances in the anaerobic fermentation of xylose by yeasts. Eur J Appl Microbiol Biotechnol 18:287–292

    Article  CAS  Google Scholar 

  26. Jeffries TW, Jin YS (2004) Metabolic engineering for improved fermentation of pentoses by yeasts. Appl Microbiol Biotechnol 63:495–509

    Article  PubMed  CAS  Google Scholar 

  27. Cornish-Bowden A, Hofmeyr JHS, Cardenas ML (1995) Strategies for manipulating metabolic fluxes in biotechnology. Bioorg Chem 23:439–449

    Article  CAS  Google Scholar 

  28. Eliasson A, Hofmeyr JHS, Pedler S, Hahn-Hägerdal B (2001) The xylose reductase/xylitol dehydrogenase/xylulokinase ratio affects product formation in recombinant xylose-utilizing Saccharomyces cerevisiae. Enzyme Microb Technol 29:288–297

    Article  CAS  Google Scholar 

  29. De Robichion-Szulmajster H (1958) Induction of enzymes of galactose pathway in mutants of Saccharomyces cerevisiae. Science 127:19–28

    Article  Google Scholar 

  30. Bhat PJ, Murthy TVS (2001) Transcriptional control of the GAL/MEL regulation of yeast Saccharomyces cerevisiae: mechanism of galactose mediated signal transduction. Mol Microbiol 40:1059–1066

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was granted by Prominent Youthfund Project of Sichuan Agricultural University with grant. No. 00924201.

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Authors and Affiliations

  1. Institute of Animal Nutrition, Sichuan Agricultural University, Ya’an, 625014, Sichuan, People’s Republic of China

    He Jun & Cai Jiayi

  2. Key Laboratory of Animal Disease-Resistance Nutrition, Ministry of Education, Ya’an, Sichuan, People’s Republic of China

    He Jun

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  1. He Jun
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  2. Cai Jiayi
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Correspondence to He Jun.

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Jun, H., Jiayi, C. Metabolic engineering of Saccharomyces cerevisiae for increased bioconversion of lignocellulose to ethanol. Indian J Microbiol 52, 442–448 (2012). https://doi.org/10.1007/s12088-012-0259-x

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  • DOI: https://doi.org/10.1007/s12088-012-0259-x

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