Differential expression of Thetrichoderma reesei β-xylanase II (xyn2) gene in the xylose-fermenting yeast Pichia stipitis

Simple item page
dc.contributor.authorDen Haan R.
dc.contributor.authorVan Zyl W.H.
dc.date.accessioned2011-05-15T15:56:37Z
dc.date.available2011-05-15T15:56:37Z
dc.date.issued2001
dc.description.abstractThe transcriptional control of two native promoters and one heterologous promoter and the production of a heterologous protein from these promoters were evaluated in the xylose-fermenting yeast Pichia stipitis cultivated on xylose and glucose as carbon sources, using the β-xylanase II xyn2 gene of Trichoderma reesei. The xyn2 gene open reading frame was fused to the P. stipitis xylose reductase gene (XYL1) promoter, the P. stipitis transketolase gene (TKL) promoter and the Saccharomyces cerevisiae phosphoglycerate kinase gene (PGK1) promoter DNA sequences on episomal plasmids. The plasmids were transformed into Pichia stipitis and gene expression and β-xylanase production monitored. The XYL1 promoter was shown to be inducible in the presence of xylose, as xyn2 transcription and β-xylanase activity could be measured when the recombinant strain was cultivated on xylose but not when it was cultivated on glucose. TKL promoter expression was found to be constitutive when either glucose or xylose was used as sole carbon source. The PGK1 promoter did not promote xyn2 transcription in P. stipitis. The molecular size of the recombinant Xyn2 protein produced by P. stipitis was 20.7 kDa, which is similar to that of the native T. reesei Xyn2 protein. This indicates no or minimal glycosylation of the recombinant protein. The recombinant xyn2-expressing strain also yielded twice the amount of biomass yielded by the control strain when cultivated in medium containing 1% birchwood xylan as sole carbon source.
dc.description.versionArticle
dc.identifier.citationApplied Microbiology and Biotechnology
dc.identifier.citation57
dc.identifier.citation4
dc.identifier.issn1757598
dc.identifier.other10.1007/s002530100790
dc.identifier.urihttp://hdl.handle.net/10019.1/9948
dc.subjectcarbon
dc.subjectfungal protein
dc.subjectphosphoglycerate kinase
dc.subjecttransketolase
dc.subjectxylan
dc.subjectxylan endo 1,3 beta xylosidase
dc.subjectxylose
dc.subjectaldehyde reductase
dc.subjectendo-1,3-beta-xylanase
dc.subjectrecombinant protein
dc.subjectxylan 1,4 beta xylosidase
dc.subjectgene
dc.subjectarticle
dc.subjectbiomass
dc.subjectcontrolled study
dc.subjectculture medium
dc.subjectDNA sequence
dc.subjectenzyme activity
dc.subjectenzyme synthesis
dc.subjectepisome
dc.subjectgene expression
dc.subjectglycosylation
dc.subjectmolecular size
dc.subjectnonhuman
dc.subjectopen reading frame
dc.subjectPichia stipitis
dc.subjectplasmid
dc.subjectpromoter region
dc.subjectSaccharomyces cerevisiae
dc.subjectTrichoderma reesei
dc.subjectyeast
dc.subjectfermentation
dc.subjectgenetics
dc.subjectgrowth, development and aging
dc.subjectmetabolism
dc.subjectpH
dc.subjectPichia
dc.subjectpolyacrylamide gel electrophoresis
dc.subjectHypocrea jecorina
dc.subjectPichia stipitis
dc.subjectSaccharomyces cerevisiae
dc.subjectTrichoderma
dc.subjectAldehyde Reductase
dc.subjectElectrophoresis, Polyacrylamide Gel
dc.subjectFermentation
dc.subjectHydrogen-Ion Concentration
dc.subjectPhosphoglycerate Kinase
dc.subjectPichia
dc.subjectPromoter Regions (Genetics)
dc.subjectRecombinant Proteins
dc.subjectTransketolase
dc.subjectXylose
dc.subjectXylosidases
dc.titleDifferential expression of Thetrichoderma reesei β-xylanase II (xyn2) gene in the xylose-fermenting yeast Pichia stipitis
dc.typeArticle
Files
Collections
Stellenbosch University - Scopus Publications