The effect of polysaccharide-degrading wine yeast transformants on the efficiency of wine processing and wine flavour.
Commercial polysaccharase preparations are applied to winemaking to improve wine processing and quality. Expression of polysaccharase-encoding genes in Saccharomyces cerevisiae allows for the recombinant strains to degrade polysaccharides that traditional commercial yeast strains cannot. In this study, we constructed recombinant wine yeast strains that were able to degrade the problem-causing grape polysaccharides, glucan and xylan, by separately integrating the Trichoderma reesei XYN2 xylanase gene construct and the Butyrivibrio fibrisolvens END1 glucanase gene cassette into the genome of the commercial wine yeast strain S. cerevisiae VIN13. These genes were also combined in S. cerevisiae VIN13 under the control of different promoters. The strains that were constructed were compared under winemaking conditions with each other and with a recombinant wine yeast strain expressing the endo-beta-1,4-glucanase gene cassette (END1) from B. fibrisolvens and the endo-beta-1,4-xylanase gene cassette (XYN4) from Aspergillus niger, a recombinant strain expressing the pectate lyase gene cassette (PEL5) from Erwinia chrysanthemi and the polygalacturonase-encoding gene cassette (PEH1) from Erwinia carotovora. Wine was made with the recombinant strains using different grape cultivars. Fermentations with the recombinant VIN13 strains resulted in significant increases in free-flow wine when Ruby Cabernet must was fermented. After 6 months of bottle ageing significant differences in colour intensity and colour stability could be detected in Pinot Noir and Ruby Cabernet wines fermented with different recombinant strains. After this period the volatile composition of Muscat d'Alexandria, Ruby Cabernet and Pinot Noir wines fermented with different recombinant strains also showed significant differences. The Pinot Noir wines were also sensorial evaluated and the tasting panel preferred the wines fermented with the recombinant strains.
Louw
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Institute for Wine Biotechnology, Department of Viticulture and Oenology, Stellenbosch University, Matieland (Stellenbosch), ZA 7602, South Africa.
La Grange
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Pretorius
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van Rensburg
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D002849:Chromatography, Gas; D003001:Cloning, Molecular; D016974:Dickeya chrysanthemi / Q000235:genetics; D005285:Fermentation / Q000235:genetics; D005511:Food Handling; D015870:Gene Expression; D005800:Genes, Fungal / Q000502:physiology; D007218:Industrial Microbiology / Q000379:methods; D016254:Mutagenesis, Insertional / Q000379:methods; D030781:Organisms, Genetically Modified / Q000235:genetics*; D016973:Pectobacterium carotovorum / Q000235:genetics; D011096:Polygalacturonase / Q000235:genetics; D011134:Polysaccharides / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic; D014920:Wine / Q000032:analysis / Q000382:microbiology*; D043325:Xylan Endo-1,3-beta-Xylosidase / Q000235:genetics; D014995:Xylosidases / Q000235:genetics; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D011134:Polysaccharides; D011994:Recombinant Proteins; D014995:Xylosidases; D011096:Polygalacturonase; D043325:Xylan Endo-1,3-beta-Xylosidase