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2-Deoxyglucose as a selective agent for derepressed mutants of Pichia stipitis

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Abstract

The glucose analog 2-deoxyglucose (2-DOG) has been used to obtain mutants depressed 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 33g/L ethanol in 45h 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.

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Abbreviations

μ:

Specific growth rate (/h)

Rs:

volumetric sugar uptake rate (g/L/h)

Qs:

specific sugar uptake rate (g/g/h)

Rpe:

volumetric ethanol production rate (g/L/h)

Qpe:

specific ethanol production rate (g/g/h)

Ye:

ethanol yield (g/g)

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Authors and Affiliations

  1. Forest Products Laboratory, Institute of Microbial Biochemical Technology, One Gifford Pinchot Drive, 53705, Madison, WI

    Hassan K. Sreenath & Thomas W. Jeffries

  2. Department of Bacteriology, University of Wisconsin-Madison, Madison, WI

    Thomas W. Jeffries

Authors
  1. Hassan K. Sreenath
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  2. Thomas W. Jeffries
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Correspondence to Thomas W. Jeffries.

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The Forest Products Laboratory is maintained in cooperation with the University of Wisconsin-Madison. This article was prepared by U.S. Goverment employees on official time, and it is therefore in the public domain and not subject to copyright.

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Sreenath, H.K., Jeffries, T.W. 2-Deoxyglucose as a selective agent for derepressed mutants of Pichia stipitis . Appl Biochem Biotechnol 77, 211–222 (1999). https://doi.org/10.1385/ABAB:77:1-3:211

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  • DOI: https://doi.org/10.1385/ABAB:77:1-3:211

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