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

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dc.contributor.author La Grange, Daniel C.
dc.contributor.author Pretorius, Isak S.
dc.contributor.author Van Zyl, Willem H.
dc.date.accessioned 2011-04-07T08:23:01Z
dc.date.available 2011-04-07T08:23:01Z
dc.date.issued 1997-07
dc.identifier.citation La 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.pdf en_ZA
dc.identifier.issn 1432-0614 (Online Version)
dc.identifier.issn 0175-7598 (Print Version)
dc.identifier.other DOI: 10.1007/s002530050924
dc.identifier.uri http://hdl.handle.net/10019.1/8481
dc.description The original publication is available at www.springerlink.com.
dc.description Includes bibliography.
dc.description.abstract A 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.extent p. 262-266 : ill.
dc.language.iso en_US en_ZA
dc.publisher Springer-Verlag en_ZA
dc.subject Bacillus en_ZA
dc.subject Saccharomyces cerevisiae en_ZA
dc.subject Pumilus en_ZA
dc.subject Xylosidase en_ZA
dc.subject Recombinant en_ZA
dc.subject Optimum en_ZA
dc.title Cloning of the Bacillus pumilus beta-xylosidase gene (xynB ) and its expression in Saccharomyces cerevisiae en_ZA
dc.type Article en_ZA
dc.rights.holder Springer-Verlag en_ZA


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