ITEM VIEW

Cloning of the Bacillus pumilus beta-xylosidase gene (xynB ) and its expression in Saccharomyces cerevisiae

dc.contributor.authorLa Grange, Daniel C.
dc.contributor.authorPretorius, Isak S.
dc.contributor.authorVan Zyl, Willem H.
dc.date.accessioned2011-04-07T08:23:01Z
dc.date.available2011-04-07T08:23:01Z
dc.date.issued1997-07
dc.identifier.citationLa Grange, D.C., Pretorius, I.S., Van Zyl, W.S. 1997. Cloning of the Bacillus pumilus b-xylosidase gene (xynB )and its expression in Saccharomyces cerevisiae. Applied Microbiology and Biotechnology, 47:262-266,DOI:10.1007/s002530050924,http://www.springerlink.com.ez.sun.ac.za/content/njwvhaqd5r6f3x7h/fulltext.pdfen_ZA
dc.identifier.issn1432-0614 (Online Version)
dc.identifier.issn0175-7598 (Print Version)
dc.identifier.otherDOI: 10.1007/s002530050924
dc.identifier.urihttp://hdl.handle.net/10019.1/8481
dc.descriptionThe original publication is available at www.springerlink.com.
dc.descriptionIncludes bibliography.
dc.description.abstractA genomic DNA library of the bacterium Bacillus pumilus PLS was constructed and the beta-xylosidase gene (xynB) was amplified from a 3-kb genomic DNA fragment with the aid of the polymerase chain reaction technique. The amplified xynB gene was inserted between the yeast alcohol dehydrogenase II gene promoter (ADH2P) and terminator (ADH2T) sequences on a multicopy episomal plasmid (pDLG11). The xynB gene was also fused in-frame to the secretion signal sequence of the yeast mating pheromone a-factor (MFa1S) before insertion between the ADH2P and ADH2T sequences on a similar multicopy episomal plasmid (pDLG12). The resulting construct ADH2P-MFa1SxynB-ADH2T was designated XLO1. Both plasmids pDLG11 and pDLG12 were introduced into Saccharomyces cerevisiae but only the expression of the XLO1 gene yielded biologically functional beta-xylosidase. The total beta-xylosidase activity remained cell-associated with a maximum activity of 0.09 nkat/ml obtained when the recombinant S. cerevisiae strain was grown for 143 h in synthetic medium. The temperature and pH optima of the recombinant Xlo1 enzyme were 45±50 °C and pH 6.6 respectively. The enzyme was thermostable at 45 °C; however, at 60 °C most of the Xlo1 was inactive after 5 min.en_ZA
dc.format.extentp. 262-266 : ill.
dc.language.isoen_USen_ZA
dc.publisherSpringer-Verlagen_ZA
dc.subjectBacillusen_ZA
dc.subjectSaccharomyces cerevisiaeen_ZA
dc.subjectPumilusen_ZA
dc.subjectXylosidaseen_ZA
dc.subjectRecombinanten_ZA
dc.subjectOptimumen_ZA
dc.titleCloning of the Bacillus pumilus beta-xylosidase gene (xynB ) and its expression in Saccharomyces cerevisiaeen_ZA
dc.typeArticleen_ZA
dc.rights.holderSpringer-Verlagen_ZA


Files in this item

Thumbnail
Thumbnail

This item appears in the following Collection(s)

ITEM VIEW