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2-Deoxyglucose as a selective agent for derepressed mutants of Pichia stipitis
Authors:Hassan K Sreenath  Thomas W Jeffries
Institution:(1) Forest Products Laboratory, Institute of Microbial Biochemical Technology, One Gifford Pinchot Drive, 53705 Madison, WI;(2) Department of Bacteriology, University of Wisconsin-Madison, Madison, WI
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. 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.
Keywords:Pichia stipitis            mutation  selection  2-deoxyglucose (2-DOG)  fermentation  ethanol  glucose repression
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