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Isolation and characterization of the Pichia stipitis xylitol dehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant

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Summary

A P. stipitis cDNA library in λgt11 was screened using antisera against P. stipitis xylose reductase and xylitol dehydrogenase, respectively. The resulting cDNA clones served as probes for screening a P. stipitis genomic library. The genomic XYL2 gene was isolated and the nucleotide sequence of the 1089 bp structural gene, and of adjacent non-coding regions, was determined. The XYL2 open-reading frame codes for a protein of 363 amino acids with a predicted molecular mass of 38.5 kDa. The XYL2 gene is actively expressed in S. cerevisiae transformants. S. cerevisiae cells transformed with a plasmid, pRD1, containing both the xylose reductase gene (XYL1) and the xylitol dehydrogenase gene (XYL2), were able to grow on xylose as a sole carbon source. In contrast to aerobic glucose metabolism, S. cerevisiae XYL1-XYL2 transformants utilize xylose almost entirely oxidatively.

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References

  • Alexander NJ (1986) Appl Microbiol Biotechnol 25:203–207

    Google Scholar 

  • Amore R, Wilhelm M, Hollenberg CP (1989) Appl Microbiol Biotechnol 30:351–357

    Google Scholar 

  • Barnett JA (1976) The utilization of sugars by yeasts. In: Tipson RS, Horton D (eds) Advances in carbohydrate chemistry and biochemistry. Academic Press, New York, pp 125–235

    Google Scholar 

  • Batt CA, Carvallo S, Easson DD, Akedo MJr (1986) Biotechnol Bioeng 28:549–553

    Google Scholar 

  • Bennetzen JL, Hall BD (1982) J Biol Chem 257:3026–3031

    Google Scholar 

  • Bergmeyer HU, Bernt E (1974) D-Glucose, Bestimmung mit Glucose-Oxydase und Peroxydase. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse. Verlag Chemie, Weinheim, 3rd edn., pp 1250–1259

    Google Scholar 

  • Bernt E, Gutmann I (1974) Äthanol, Bestimmung mit Alkohol-Dehydrogenase und NAD. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse. Verlag Chemie, Weinheim, 3rd edn., pp 1545–1548

    Google Scholar 

  • Beutler H-O (1984) Xylitol. In: Bergmeyer HU (ed) Methods of enzymatic analysis, Volume VI, Metabolites 1: Carbohydrates. Verlag Chemie, Weinheim, 3rd edn., pp 484–490

    Google Scholar 

  • Bisson LF (1988) J Bacteriol 170:4838–4845

    Google Scholar 

  • Blake MS, Johnston KH, Russel-Jones GJ, Gottschlich EC (1984) Anal Biochem 136:175–179

    Google Scholar 

  • Broach JR (1983) Methods Enymol 101:307–325

    Google Scholar 

  • Bruinenberg PM, De Bot PHM, Van Dijken JP, Scheffers WA (1983a) Eur J Appl Microbiol Biotechnol 18:287–292

    Google Scholar 

  • Bruinenberg PM, Van Dijken JP, Scheffers WA (1983b) J Gen Microbiol 129:965–971

    Google Scholar 

  • Bruinenberg PM, De Bot PHM, Van Dijken JP, Scheffers WA (1984) Appl Microbiol Biotechnol 19:256–260

    Google Scholar 

  • Ciriacy M, Porep H (1986) Conversion of pentoses to ethanol by baker's yeast. In: Magnien E (ed) Biomolecular engineering in the european community. Martinus Nijhoff Publishers, Dordrecht, pp 675–681

    Google Scholar 

  • Cirillo VP (1968) J Bacteriol 95:603–611

    Google Scholar 

  • Dellweg H, Rizzi M, Mether H, Debus D (1984) Biotechnol Lett 6:395–400

    Google Scholar 

  • Dobson MJ, Tuite MF, Roberts NA, Kingsman AJ, Kingsman SM, Perkins RE, Conroy SC, Dunbar B, Fothergill LA (1982) Nucleic Acids Res 10:2625–2637

    Google Scholar 

  • Does AL, Bisson LF (1989) Appl Environ Microbiol 55:159–164

    Google Scholar 

  • Gong C-S, Chen LF, Flickinger MC, Chiang L-C, Tsao GT (1981) Appl Environ Microbiol 41:430–436

    Google Scholar 

  • Gong C-S, Claypool TA, McCracken LD, Maun CH, Ueng PP, Tsao GT (1983) Biotechnol Bioeng 25:85–102

    Google Scholar 

  • Gubler U, Hoffman BJ (1983) Gene 25:263–269

    Google Scholar 

  • Hagedorn J, Ciriacy M (1989) Curr Genet 16:27–33

    Google Scholar 

  • Halliwell G, Lovelady J (1981) J Gen Microbiol 126:211–217

    Google Scholar 

  • Hamilton R, Watanabe CK, de Boer HA (1987) Nucleic Acids Res 15:3581–3591

    Google Scholar 

  • Henikoff S (1984) Gene 28:351–359

    Google Scholar 

  • Hohn B, Murray K (1977) Proc Nat Acad Sci USA 74:3259–3263

    Google Scholar 

  • Huynh TV, Young RA, Davis RW (1985) Construction and screening cDNA libraries in λgt10 and λgt11. In: Glover D (ed) DNA cloning, a practical approach. IRL Press, Oxford, pp 49–78

    Google Scholar 

  • Jeffries TW (1983) Adv Biochem Eng 27:1–32

    Google Scholar 

  • Jeffries TW (1985) TIB 3:208–212

    Google Scholar 

  • Johnston M, Davis RW (1984) Mol Cell Biol 4:1440–1448

    Google Scholar 

  • Kilian SG, van Uden N (1988) Appl Microbiol Biotechnol 27:545–548

    Google Scholar 

  • Kotyk A (1967) Folia Microbiol 12:121–131

    Google Scholar 

  • Kozak M (1981) Nucleic Acids Res 9:5233–5252

    Google Scholar 

  • Laemmli UK (1970) Nature 227:680–685

    Google Scholar 

  • Lagunas R, Gancedo JM (1973) Eur J Biochem 37:90–94

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Mierendorf RC, Percy C, Young RA (1987) Methods Enymol 152:458–469

    Google Scholar 

  • Porep HJ (1987) Xyluloseverwertung bei Saccharomyces cerevisiae. PhD thesis, University of Düsseldorf, Federal Republic of Germany

  • Proudfoot NJ, Brownlee GG (1976) Nature 263:211–214

    Google Scholar 

  • Ramos J, Szkutnicka K, Cirillo VP (1988) J Bacteriol 170:5375–5377

    Google Scholar 

  • Rizzi M, Erlemann P, Bui-Thanh N-A, Dellweg H (1988) Appl Microbiol Biotechnol 29:148–154

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) Proc Nat Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sarthy AV, McConaughy BL, Lobo Z, Sundstrom JA, Furlong CE, Hall BD (1987) Appl Environ Microbiol 53:1996–2000

    Google Scholar 

  • Senac T, Hahn-Hägerdal B (1990) Appl Environ Microbiol 56:120–126

    Google Scholar 

  • Skoog K, Hahn-Hägerdal B (1988) Enzyme Microb Technol 10:66–80

    Google Scholar 

  • Toivola A, Yarrow D, Van Den Bosch E, Van Dijken JP, Scheffers WA (1984) Appl Environ Microbiol 47:1221–1223

    Google Scholar 

  • Towbin H, Staelin T, Gordon J (1979) Proc Natl Acad Sci USA 76:4350–5354

    Google Scholar 

  • Ueng PP, Hunter CA, Gong C-S, Tsao GT (1981) Biotechnol Lett 3:315–320

    Google Scholar 

  • Verduyn C, van Kleef R, Frank J, Schreuder H, van Dijken JP, Scheffers WA (1985) Biochem J 226:669–677

    Google Scholar 

  • Wang PY, Schneider H (1980) Can J Microbiol 26:1165–1168

    Google Scholar 

  • Wieland OH (1984) Glycerol, UV-method. In: Bergmeyer HU (ed) Methods of enzymatic analysis, volume VI, Metabolites 1: Carbohydrates. Verlag Chemie, Weinheim, 3rd edn., pp 504–510

    Google Scholar 

  • Wierenga RK, De Maeyer MCH, Hol WGJ (1985) Biochemistry 24:1346–1357

    Google Scholar 

  • Wierenga RK, Terpstra P, Hol WGJ (1986) J Mol Biol 187:101–107

    Google Scholar 

  • Zalkin H, Yanofsky C (1982) J Biol Chem 257:1491–1500

    Google Scholar 

  • Zamenhoff S (1958) Methods Enymol 3:696–704

    Google Scholar 

  • Zaret KS, Sherman F (1982) Cell 28:563–573

    Google Scholar 

Download references

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Authors and Affiliations

  1. Institut für Mikrobiologie, Heinrich-Heine-Universität, Universitätstrasse 1, W-4000, Düsseldorf, Germany

    Peter Kötter, René Amore, Cornelis P. Hollenberg & Michael Ciriacy

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  1. Peter Kötter
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  2. René Amore
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  3. Cornelis P. Hollenberg
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  4. Michael Ciriacy
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Communicated by F.K. Zimmermann

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Kötter, P., Amore, R., Hollenberg, C.P. et al. 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 (1990). https://doi.org/10.1007/BF00327019

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  • DOI: https://doi.org/10.1007/BF00327019

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