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Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation

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Journal of Industrial Microbiology and Biotechnology

Abstract

A new method for the selection of Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability to ferment xylose to ethanol was developed. The method is based on the ability of P. stipitis and H. polymorpha colonies to grow and produce ethanol on agar plates with xylose as the sole carbon and energy source. Secreted ethanol, in contrast to xylose, supports growth of cells of the indicator xylose-negative strains (the wild-type strain of Saccharomyces cerevisiae or Δxyl1 mutant of H. polymorpha) mixed with agar medium. The size of the tester culture-growth zone around xylose-grown colonies appeared to be dependent on the amount of secreted ethanol. Mutants with altered (decreased or elevated) ethanol production in xylose medium have been isolated using this method. The mutants exhibited pleiotropic alterations in enzymatic activities of the intermediary xylose metabolism.

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Acknowledgments

Authors are grateful to Dr. O.V. Stasyk for critical reading the manuscript and to Mrs B.V. Kshanovska (both from Institute of Cell Biology, NAS of Ukraine, Lviv) for participation in isolation of H. polymorpha mutants. The work was supported in part by the Polish National research grant KBN 3 PO4B 003 23 and the INTAS grant Nr 05-1000005-7730.

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Authors and Affiliations

  1. Department of Metabolic Engineering, Rzeszów University, Ćwiklińskiej 2, 35-601, Rzeszów, Poland

    Dorota Grabek-Lejko, Bernadetta Oklejewicz & Andriy A. Sibirny

  2. Institute of Cell Biology, NAS of Ukraine, Drahomanov Street, 14/16, Lviv, 79005, Ukraine

    Olena B. Ryabova, Andriy Y. Voronovsky & Andriy A. Sibirny

Authors
  1. Dorota Grabek-Lejko
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  2. Olena B. Ryabova
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  3. Bernadetta Oklejewicz
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  4. Andriy Y. Voronovsky
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  5. Andriy A. Sibirny
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Correspondence to Andriy A. Sibirny.

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Grabek-Lejko, D., Ryabova, O.B., Oklejewicz, B. et al. Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation. J Ind Microbiol Biotechnol 33, 934–940 (2006). https://doi.org/10.1007/s10295-006-0147-7

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  • DOI: https://doi.org/10.1007/s10295-006-0147-7

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