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Establishment of l-arabinose fermentation in glucose/xylose co-fermenting recombinant Saccharomyces cerevisiae 424A(LNH-ST) by genetic engineering

  • Applied Genetics and Molecular Biotechnology
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Abstract

Cost-effective and efficient ethanol production from lignocellulosic materials requires the fermentation of all sugars recovered from such materials including glucose, xylose, mannose, galactose, and l-arabinose. Wild-type strains of Saccharomyces cerevisiae used in industrial ethanol production cannot ferment d-xylose and l-arabinose. Our genetically engineered recombinant S. cerevisiae yeast 424A(LNH-ST) has been made able to efficiently ferment xylose to ethanol, which was achieved by integrating multiple copies of three xylose-metabolizing genes. This study reports the efficient anaerobic fermentation of l-arabinose by the derivative of 424A(LNH-ST). The new strain was constructed by over-expression of two additional genes from fungi l-arabinose utilization pathways. The resulting new 424A(LNH-ST) strain exhibited production of ethanol from l-arabinose, and the yield was more than 40%. An efficient ethanol production, about 72.5% yield from five-sugar mixtures containing glucose, galactose, mannose, xylose, and arabinose was also achieved. This co-fermentation of five-sugar mixture is important and crucial for application in industrial economical ethanol production using lignocellulosic biomass as the feedstock.

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References

  • Barnett JA (1976) The utilization of sugars by yeasts. Adv Carbohydr Chem Biochem 32:125–234

    Article  CAS  Google Scholar 

  • Becker DM, Guarente L (1991) High-efficiency transformation of yeast by electroporation. Meth Enzymol 194:182–187

    Article  CAS  Google Scholar 

  • Becker J, Boles E (2003) A modified Saccharomyces cerevisiae strain that consumes l-arabinose and produces ethanol. Appl Environ Microbiol 69:4144–4150

    Article  CAS  Google Scholar 

  • Bettiga M, Bengtsson O, Hahn-Hagerdal B, Gorwa-Grauslund MF (2009) Arabinose and xylose fermentation by recombinant Saccharomyces cerevisiae expressing a fungal pentose utilization pathway. Microb Cell Fact 8:40

    Article  Google Scholar 

  • Chevallier MR, Aigle M (1979) Qualitative detection of penicillinase produced by yeast strains carrying chimeric yeast–coli plasmids. FEBS Lett 108:179–180

    Article  CAS  Google Scholar 

  • Dien BS, Kurtzman CP, Saha BC, Bothast RJ (1996) Screening for l-arabinose fermenting yeasts. Appl Biochem Biotechnol 57–58:233–242

    Article  Google Scholar 

  • Eliasson A, Christensson C, Wahlbom CF, Hahn-Hagerdal B (2000) Anaerobic xylose fermentation by recombinant Saccharomyces cerevisiae carrying XYL1, XYL2, and XKS1 in mineral medium chemostat cultures. Appl Environ Microbiol 66:3381–3386

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Ho NW, Chen Z, Brainard AP (1998) Genetically engineered Saccharomyces yeast capable of effective cofermentation of glucose and xylose. Appl Environ Microbiol 64:1852–1859

    CAS  Google Scholar 

  • Ho NW, Chen Z, Brainard AP, Sedlak M (1999) Successful design and development of genetically engineered Saccharomyces yeasts for effective cofermentation of glucose and xylose from cellulosic biomass to fuel ethanol. Adv Biochem Eng Biotechnol 65:163–192

    CAS  Google Scholar 

  • Horvath IT, Anastas PT (2007) Innovations and green chemistry. Chem Rev 107:2169–2173

    Article  CAS  Google Scholar 

  • Kotter P, Amore R, Hollenberg CP, Ciriacy M (1990) Isolation and characterization of the Pichia stipitis xylitol dehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant. Curr Genet 18:493–500

    Article  CAS  Google Scholar 

  • Kou SC, Christensen MS, Cirillo VP (1970) Galactose transport in Saccharomyces cerevisiae. II. Characteristics of galactose uptake and exchange in galactokinaseless cells. J Bacteriol 103:671–678

    CAS  Google Scholar 

  • Kurtzman CP, Dien BS (1998) Candida arabinofermentans, a new l-arabinose fermenting yeast. Antonie Leeuwenhoek 74:237–243

    Article  CAS  Google Scholar 

  • Kuyper M, Winkler AA, van Dijken JP, Pronk JT (2004) Minimal metabolic engineering of Saccharomyces cerevisiae for efficient anaerobic xylose fermentation: a proof of principle. FEMS Yeast Res 4:655–664

    Article  CAS  Google Scholar 

  • Kuyper M, Hartog MMP, Toirkens MJ, Almering MJH, Winkler AA, van Dijken JP, Pronk JT (2005) Metabolic engineering of a xylose-isomerase-expressing Saccharomyces cerevisiae strain for rapid anaerobic xylose fermentation. FEMS Yeast Res 5:399–409

    Article  CAS  Google Scholar 

  • Matsushika A, Inoue H, Kodaki T, Sawayama S (2009) Ethanol production from xylose in engineered Saccharomyces cerevisiae strains: current state and perspectives. Appl Microbiol Biotechnol 84:37–53

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Metz B, de Vries RP, Polak S, Seidl V, Seiboth B (2009) The Hypocrea jecorina (syn. Trichoderma reesei) lxr1 gene encodes a d-mannitol dehydrogenase and is not involved in l-arabinose catabolism. FEBS Lett 583:1309–1313

    Article  CAS  Google Scholar 

  • Pail M, Peterbauer T, Seiboth B, Hametner C, Druzhinina I, Kubicek CP (2004) The metabolic role and evolution of l-arabinitol 4-dehydrogenase of Hypocrea jecorina. Eur J Biochem 271:1864–1872

    Article  CAS  Google Scholar 

  • Richard P, Londesborough J, Putkonen M, Kalkkinen N, Penttila M (2001) Cloning and expression of a fungal l-arabinitol 4-dehydrogenase gene. J Biol Chem 276:40631–40637

    Article  CAS  Google Scholar 

  • Richard P, Verho R, Putkonen M, Londesborough J, Penttila M (2003) Production of ethanol from l-arabinose by Saccharomyces cerevisiae containing a fungal l-arabinose pathway. FEMS Yeast Res 3:185–189

    Article  CAS  Google Scholar 

  • Sedlak M, Ho NW (2001) Expression of E. coli araBAD operon encoding enzymes for metabolizing l-arabinose in Saccharomyces cerevisiae. Enzyme Microb Technol 28:16–24

    Article  CAS  Google Scholar 

  • Sedlak M, Ho NW (2004a) Characterization of the effectiveness of hexose transporters for transporting xylose during glucose and xylose co-fermentation by a recombinant Saccharomyces yeast. Yeast 21:671–684

    Article  CAS  Google Scholar 

  • Sedlak M, Ho NW (2004b) Production of ethanol from cellulosic biomass hydrolysates using genetically engineered Saccharomyces yeast capable of cofermenting glucose and xylose. Appl Biochem Biotechnol 113–116:403–416

    Article  Google Scholar 

  • Sonderegger M, Sauer U (2003) Evolutionary engineering of Saccharomyces cerevisiae for anaerobic growth on xylose. Appl Environ Microbiol 69:1990–1998

    Article  CAS  Google Scholar 

  • Taketo A (1988) DNA transfection of Escherichia coli by electroporation. Biochim Biophys Acta 949:318–324

    CAS  Google Scholar 

  • Toivari MH, Aristidou A, Ruohonen L, Penttila M (2001) Conversion of xylose to ethanol by recombinant Saccharomyces cerevisiae: importance of xylulokinase (XKS1) and oxygen availability. Metab Eng 3:236–249

    Article  CAS  Google Scholar 

  • Verho R, Londesborough J, Penttila M, Richard P (2003) Engineering redox cofactor regeneration for improved pentose fermentation in Saccharomyces cerevisiae. Appl Environ Microbiol 69:5892–5897

    Article  CAS  Google Scholar 

  • Verho R, Putkonen M, Londesborough J, Penttila M, Richard P (2004) A novel NADH-linked l-xylulose reductase in the l-arabinose catabolic pathway of yeast. J Biol Chem 279:14746–14751

    Article  CAS  Google Scholar 

  • Watanabe S, Kodaki T, Makino K (2006) Cloning, expression, and characterization of bacterial l-arabinose l-dehydrogenase involved in an alternative pathway of l-arabinose metabolism. J Biol Chem 281:2612–2623

    Article  CAS  Google Scholar 

  • Wisselink HW, Toirkens MJ, del Rosario Franco Berriel M, Winkler AA, van Dijken JP, Pronk JT, van Maris AJA (2007) Engineering of Saccharomyces cerevisiae for efficient anaerobic alcoholic fermentation of l-arabinose. Appl Environ Microbiol 73:4881–4891

    Article  CAS  Google Scholar 

  • Wisselink HW, Toirkens MJ, Wu Q, Pronk JT, van Maris AJ (2009) Novel evolutionary engineering approach for accelerated utilization of glucose, xylose, and arabinose mixtures by engineered Saccharomyces cerevisiae strains. Appl Environ Microbiol 75:907–914

    Article  CAS  Google Scholar 

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Acknowledgement

The project was financially supported by the US Department of Energy Biomass Program, Contract G017059-16649.

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Authors and Affiliations

  1. Laboratory of Renewable Resources Engineering, Purdue University, West Lafayette, IN, USA

    Aloke Kumar Bera, Miroslav Sedlak, Aftab Khan & Nancy W. Y. Ho

  2. Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN, USA

    Miroslav Sedlak

  3. School of Chemical Engineering, Purdue University, West Lafayette, IN, USA

    Nancy W. Y. Ho

Authors
  1. Aloke Kumar Bera
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  2. Miroslav Sedlak
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  3. Aftab Khan
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  4. Nancy W. Y. Ho
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Correspondence to Nancy W. Y. Ho.

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Bera, A.K., Sedlak, M., Khan, A. et al. Establishment of l-arabinose fermentation in glucose/xylose co-fermenting recombinant Saccharomyces cerevisiae 424A(LNH-ST) by genetic engineering. Appl Microbiol Biotechnol 87, 1803–1811 (2010). https://doi.org/10.1007/s00253-010-2609-0

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  • DOI: https://doi.org/10.1007/s00253-010-2609-0

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