Generation of the improved recombinant xylose-utilizing Saccharomyces cerevisiae TMB 3400 by random mutagenesis and physiological comparison with Pichia stipitis CBS 6054

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dc.contributor.authorWahlbom C.F.
dc.contributor.authorVan Zyl W.H.
dc.contributor.authorJonsson L.J.
dc.contributor.authorHahn-Hagerdal B.
dc.contributor.authorCordero Otero R.R.
dc.date.accessioned2011-05-15T16:03:27Z
dc.date.available2011-05-15T16:03:27Z
dc.date.issued2003
dc.description.abstractThe recombinant xylose-utilizing Saccharomyces cerevisiae TMB 3399 was constructed by chromosomal integration of the genes encoding D-xylose reductase (XR), xylitol dehydrogenase (XDH), and xylulokinase (XK). S. cerevisiae TMB 3399 was subjected to chemical mutagenesis with ethyl methanesulfonate and, after enrichment, 33 mutants were selected for improved growth on D-xylose and carbon dioxide formation in Durham tubes. The best-performing mutant was called S. cerevisiae TMB 3400. The novel, recombinant S. cerevisiae strains were compared with Pichia stipitis CBS 6054 through cultivation under aerobic, oxygen-limited, and anaerobic conditions in a defined mineral medium using only D-xylose as carbon and energy source. The mutation led to a more than five-fold increase in maximum specific growth rate, from 0.0255 h-1 for S. cerevisiae TMB 3399 to 0.14 h-1 for S. cerevisiae TMB 3400, whereas P. stipitis grew at a maximum specific growth rate of 0.44 h-1. All yeast strains formed ethanol only under oxygen-limited and anaerobic conditions. The ethanol yields and maximum specific ethanol productivities during oxygen limitation were 0.21, 0.25, and 0.30 g ethanol g xylose-1 and 0.001, 0.10, and 0.16 g ethanol g biomass-1 h-1 for S. cerevisiae TMB 3399, TMB 3400, and P. stipitis CBS 6054, respectively. The xylitol yield under oxygen-limited and anaerobic conditions was two-fold higher for S. cerevisiae TMB 3399 than for TMB 3400, but the glycerol yield was higher for TMB 3400. The specific activity, in U mg protein-1, was higher for XDH than for XR in both S. cerevisiae TMB 3399 and TMB 3400, while P. stipitis CBS 6054 showed the opposite relation. S. cerevisiae TMB 3400 displayed higher specific XR, XDH and XK activities than TMB 3399. Hence, we have demonstrated that a combination of metabolic engineering and random mutagenesis was successful to generate a superior, xylose-utilizing S. cerevisiae, and uncovered distinctive physiological properties of the mutant. © 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
dc.description.versionArticle
dc.identifier.citationFEMS Yeast Research
dc.identifier.citation3
dc.identifier.citation3
dc.identifier.issn15671356
dc.identifier.other10.1016/S1567-1356(02)00206-4
dc.identifier.urihttp://hdl.handle.net/10019.1/12629
dc.subjectalcohol
dc.subjectcarbon
dc.subjectmineral
dc.subjectunclassified drug
dc.subjectxylitol
dc.subjectxylitol dehydrogenase
dc.subjectxylose
dc.subjectxylose reductase
dc.subjectxylulokinase
dc.subjectarticle
dc.subjectbiomass
dc.subjectcarbohydrate synthesis
dc.subjectcontrolled study
dc.subjectenergy resource
dc.subjectfungal strain
dc.subjectfungus growth
dc.subjectgrowth rate
dc.subjectmetabolic engineering
dc.subjectmutagenesis
dc.subjectnonhuman
dc.subjectPichia stipitis
dc.subjectSaccharomyces cerevisiae
dc.subjectspecies differentiation
dc.subjectAerobiosis
dc.subjectAnaerobiosis
dc.subjectFermentation
dc.subjectKinetics
dc.subjectMutagenesis
dc.subjectOxygen
dc.subjectPichia
dc.subjectRecombination, Genetic
dc.subjectSaccharomyces cerevisiae
dc.subjectTransformation, Genetic
dc.subjectXylose
dc.subjectPichia
dc.subjectPichia stipitis
dc.subjectSaccharomyces
dc.subjectSaccharomyces cerevisiae
dc.titleGeneration of the improved recombinant xylose-utilizing Saccharomyces cerevisiae TMB 3400 by random mutagenesis and physiological comparison with Pichia stipitis CBS 6054
dc.typeArticle
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