The heterologous expression of polysaccharidase-encoding genes with oenological relevance in Saccharomyces cerevisiae.
OBJECTIVE
The main objective of this study was to develop polysaccharide-degrading wine strains of Saccharomyces cerevisiae, which are able to improve aspects of wine processing and clarification, as well as colour extraction and stabilization during winemaking.


RESULTS
Two yeast expression/secretion gene cassettes were constructed, namely (i) a pectinase gene cassette (pPPK) consisting of the endo-polygalacturonase gene (pelE) from Erwinia chrysanthemi and the pectate lyase gene (peh1) from Erwinia carotovora and (ii) a glucanase/xylanase gene cassette (pEXS) containing the endo-beta-1,4-glucanase gene (end1) from Butyrivibrio fibrisolvens and the endo-beta-1,4-xylanase gene (xynC) from Aspergillus niger. The commercial wine yeast strain, VIN13, was transformed separately with these two gene cassettes and checked for the production of pectinase, glucanase and xylanase activities. Pinot Noir, Cinsaut and Muscat d'Alexandria grape juices were fermented using the VIN13[pPPK] pectinase- and the VIN13[pEXS] glucanase/xylanase-producing transformants. Chemical analyses of the resultant wines indicated that (i) the pectinase-producing strain caused a decrease in the concentration of phenolic compounds in Pinot Noir whereas the glucanase/xylanase-producing strain caused an increase in phenolic compounds presumably because of the degradation of the grape skins; (ii) the glucanase/xylanase-producing strain caused a decrease in wine turbidity, especially in Pinot Noir wine, as well as a clear increase in colour intensity and (iii) in the Muscat d'Alexandria and Cinsaut wines, the differences between the control wines (fermented with the untransformed VIN3 strain) and the wines produced by the two transformed strains were less prominent showing that the effect of these polysaccharide-degrading enzymes is cultivar-dependent.


CONCLUSIONS
The recombinant wine yeasts producing pectinase, glucanase and xylanase activities during the fermentation of Pinot Noir, Cinsaut and Muscat d'Alexandria grape juice altered the chemical composition of the resultant wines in a way that such yeasts could potentially be used to improve the clarity, colour intensity and stability and aroma of wine.


CONCLUSIONS
Aspects of commercial-scale wine processing and clarification, colour extraction and stabilization, and aroma enhancement could potentially be improved by the use of polysaccharide-degrading wine yeasts without the addition of expensive commercial enzyme preparations. This offers the potential to further improve the price:quality ratio of wine according to consumer expectations.
Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Matieland, South Africa.
P
P
van Rensburg
M L A
ML
Strauss
M G
MG
Lambrechts
R R
RR
Cordero Otero
I S
IS
Pretorius
D001234:Aspergillus niger / Q000235:genetics; D045856:Butyrivibrio / Q000235:genetics; D016974:Dickeya chrysanthemi / Q000235:genetics; D005285:Fermentation; D005516:Food Microbiology; D015870:Gene Expression; D005800:Genes, Fungal; D005818:Genetic Engineering; D007218:Industrial Microbiology; D030781:Organisms, Genetically Modified; D016973:Pectobacterium carotovorum / Q000235:genetics; D011134:Polysaccharides / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014920:Wine
D011134:Polysaccharides