2-Deoxyglucose as a Selective Agent for Derepressed Mutants of Pichia stipitis

Part of the Applied Biochemistry and Biotechnology book series (ABAB)

Abstract

The glucose analog 2-deoxyglucose (2-DOG) has been used to obtain mutants derepressed for pentose metabolism. Some researchers have used 2-DOG alone whereas others have used it in the presence of a glucose-repressible carbon source. We examined both methods and screened mutant strains for improved use of xylose in the presence of glucose. Pichia stipitis mutants selected for growth on D-xylose in the presence of 2-DOG used xylose from a 1:1 glucose:xylose mixture more rapidly than did their parents. One of these mutants, FPL-DX26, completely consumed xylose in the presence of glucose and produced 33 g/L ethanol in 45 h from 80 g/L of this sugar mixture. Mutants selected for growth on 2-DOG alone did not show significant improvement. Selection for growth on D-xylose in the presence of 2-DOG has been useful in developing parental strains for further genetic manipulation.

Index Entries

Pichia stipitis mutation selection 2-deoxyglucose (2-DOG) fermentation ethanol glucose repression 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Patel, G. B. (1984), Appl. Microbiol. BioTechnol. 20, 111–117.CrossRefGoogle Scholar
  2. 2.
    Delgenes, J. P., Moletta, R., and Navarro, J. M. (1988), Appl. Microbiol. Biotechnol. 29, 155–161.Google Scholar
  3. 3.
    Dellweg, H., Klein, C, Prahl, S., Rizzi, M., and Weigert, B. (1990), Food BioTechnol. 4, 137–148.CrossRefGoogle Scholar
  4. 4.
    Jeffries, T. W. and Kurtzman, C. P. (1994), Enzyme Microb. Technol. 16, 922–932.CrossRefGoogle Scholar
  5. 5.
    Takahashi, D. F., Carvalhal, M. L., and Alterthum, F. (1994), Biotechnol. Lett. 16, 747–750.CrossRefGoogle Scholar
  6. 6.
    Alexander, N. J. (1990), Curr. Genet. 17, 493–497.CrossRefGoogle Scholar
  7. 7.
    Jeffries, T. W. (1984), Enzyme Microb. Technol. 6, 254–258.CrossRefGoogle Scholar
  8. 8.
    Lachke, A. H. and Jeffries, T. W. (1986), Enzyme Microb. Technol. 8, 353–359.CrossRefGoogle Scholar
  9. 9.
    Laplace, J. M., Delgenes, J. P., Molleta, R., and Navarro, J. M. (1992), Enzyme Microb. Technol. 14, 644–648.CrossRefGoogle Scholar
  10. 10.
    McCracken, L. D. and Gong, C. S. (1983), Adv. Biochem. Bioeng. 27, 33–85.Google Scholar
  11. 11.
    Parekh, S. R., Yu, S., and Waymann, M. (1986), Appl. Microbiol. BioTechnol. 25, 300–304.CrossRefGoogle Scholar
  12. 12.
    Zhang, M., Eddy, C, Deanda, K., Finkelstein, M., and Picataggio, S. (1995), Science 267, 240–243.CrossRefGoogle Scholar
  13. 13.
    D’Amore, T. and Dowhanick, T. M. (1994), J. Am. Soc. Brew. Chem. 52, 12–15.Google Scholar
  14. 14.
    Entian, K. D. and Fröhlich, K.-U. (1984), J. Bacteriol. 158, 29–35.Google Scholar
  15. 15.
    Lobo, Z. and Maitra, P. K. (1979), Mol. Gen. Genet. 157, 297–300.CrossRefGoogle Scholar
  16. 16.
    Jones, R. M., Russell, I., and Stuart, G. G. (1986), J. Am. Soc. Brew. Chem. 44, 161–166.Google Scholar
  17. 17.
    Goffrini, P., Ficarelli, A., and Ferrero, I. (1995), Microbiology 141, 441–447.CrossRefGoogle Scholar
  18. 18.
    Pardo, E.H., Sunayama, S., Pedrosa, F.O., and Rigo, L.U. (1992), Can. J. Microbiol. 38, 417–422.CrossRefGoogle Scholar
  19. 19.
    Sreenath, H. K. and Jeffries, T. W. (1996), Biotechnol. Tech. 10, 239–242CrossRefGoogle Scholar
  20. 20.
    Jeffries, T. W. and Livingston, P. L. (1992), U. S. Patent No. 5, 126, 266.Google Scholar
  21. 22.
    Sreenath, H. K. and Jeffries, T. W. (1997), Appl. Biochem. BioTechnol. 63/65, 109–116.CrossRefGoogle Scholar
  22. 22.
    Lu, P., Davis, B. P., Hendrick, J., and Jeffries, T. W. (1998), Appl. Microbiol BioTechnol. 49, 141–146.CrossRefGoogle Scholar
  23. 23.
    Jeffries, T. W. (1982), Biotechnol. Bioeng. Symp. 12, 103–110.Google Scholar
  24. 24.
    Verhaar, L. A. Th. and Küster, B. F. M. (1981), J. Chromatog. 210, 279–290.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  1. 1.Institute of Microbial Biochemical TechnologyForest Products LaboratoryMadisonUSA
  2. 2.Department of BacteriologyUniversity of Wisconsin-MadisonMadisonUSA

Personalised recommendations