Thesis (MAgricAdmin (Agricultural Economics))--University of Stellenbosch, 2005. / The wine industry is one of the oldest commercial activities in South Africa. The South African wine grape industry annually produces more than a million tonnes of grapes, making the country the ninth largest producer in the world. The total area under wine grape production is divided into eight regions for administrative purposes. These boundaries are a legacy of the era of controlled marketing and there is continued meaningfulness of having various classifications such as ‘wine of origin’ scheme. The purpose of this study is to identify the factors that determine the producers’ choice of wine grape cultivars in the wine regions in South Africa. Time series data for the period 1990-2003 were used to estimate the parameters of linear regression models. Two equations for each wine grape cultivar in each region were postulated and estimated using Ordinary Least Squares as applied with Eviews. Further, a stepwise regression as applied in STATISTICA was used to eliminate the parameters that were not statistically significant at five percent significant level. In identifying the factors that determine the choice of wine grape cultivars in the regions, the results showed that each wine grape cultivar in each region has its own factors influencing the producers’ choice of that specific wine grape cultivar. Same wine grape cultivars in different regions similarly have its own factors determining the producers’ choice. The implication of this is that there are differences in terms of the requirements and types of crops and wine grape cultivars grown in each region. However, the most important result that emerged with regular frequency is that, the factors determining the producers’ choice of a specific wine grape cultivar for each region is price of other wine grape cultivars and competitive products in that wine region. The price of specific wine grape cultivars only had an influence on few wine grape cultivars. The implication is that the producers in South Africa appears to consider the prices of other wine grape cultivars and competitive products before making a choice of whether to plant or uproot a specific wine grape cultivar more than the price of the specific wine grape cultivar. This supports the theory that farm prices play a key role in allocating resources and in rewarding efficient producers.

Thesis (MBA)--University of Stellenbosch, 2011. / Wine is not the alcoholic beverage of choice for the average South African. South Africa‟s wine consumption per capita is much lower than in other wine producing countries. It would be hugely beneficial to the South African wine industry if our domestic market were beter developed. This study aims to improve our understanding and knowledge of the South African wine market by exploring the purchasing behaviour and preferences of the South African wine consumer. This objective is attained by identifying whether a correlation exists between demographic variables and wine purchasing behaviour in the South African market. A literature review of international research indicated that various demographical, behavioural and other determinants significantly influence a consumer's decision to buy wine. This information was used to formulate a data collection strategy. A quota sampling strategy was used and the data was collected with the help of an online questionnaire. This study used the chi-squared test of a contingency table to determine if a relationship exists between any of the variables and the Cramer‟s V test was used to measure the strength of the association. This study produced similar results as other international studies conducted on the factors influencing wine purchasing behaviour. The male survey respondents tended to spend more on average on a bottle of wine and had on average a higher wine knowledge when compared to the female survey respondents. The older survey participants consumed more wine and tended to spend more per month on wine compared to their younger counterparts. The respondents in the higher income groups tended to spend more per month on wine and purchased wine that is more expensive when compared to their counterparts in the lower income groups. This study found significant relationships between wine knowledge and wine purchasing variables. Higher levels of wine knowledge often lead to higher wine consumption levels. Furthermore, the respondents in the higher wine expenditure groups generally possessed a higher wine knowledge compared to the respondents in the lower expenditure groups. This study found a number of significant relationships between the purchasing behaviour and the preferences of the survey respondents. The respondents in the higher consumption groups preferred red or white wine and had an adverse preference for wine in the “other” category. Furthermore, most red and sparkling wine consumers preferred a natural cork while most dry white wine consumers preferred a screwcap as closure method. The respondents displayed a possible willingness to pay more for a bottle of red wine than for a bottle of white wine. Furthermore, the respondents favoured screwcap in the lower average price category and preferred a natural cork in the higher average price categories. The survey participants tended to purchase wine at different price points and did not only buy wine in one price bracket.

Thesis (MBA)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: This mini-study project examines the strategic position of the Douglas Wine Cellar in context with the dynamics in the wine industry. For success in the future it suggests that producers, manufactures and marketing strategies will have to focus on doing the same things differently as well as doing different things, with the emphasis on improving productivity, increasing quality focus and generating value-added products aimed at meeting market requirements. The greater focus on productivity in agriculture should include wider use of effective farming practices, more effective use of capital equipment, more effective implementation of Research & Development results, product differentiation and brand building as well as the development of distribution channels with the ability to deliver these products in a cost effective and timely manner. There is no single "Superhighway". Routes are complex and discontinuous with an ever increasing pace of change. Today we live and work in a technology rich society. Rapid advancements in all industries are occurring daily. To survive and prosper, Douglas Wine Cellar must commit to a continual processof advancement and improvement of strategies to fulfill the vision of GWKLtd. THE VISION OF GWK IS TO BE A SUSTAINABLE AGRI-BUSINESS OF EXCELLENCE / AFRIKAANSE OPSOMMING: Hierdie werkstuk is onderneem ten einde die strategiese posisie van Douglas Wynkelder in die konteks van die wynindustrie dinamika te ondersoek. Sukses in die toekoms hang daarvan af of produsente, vervaardigers en bemarkingstrategië daarop fokus om dieselfde dinge verskillend te doen sowel as om nuwe stelsels te implementeer, met die klem op produktiwiteit, verhoogde kwaliteit en die generering van waardetoegevoegde produkte wat aan die markbehoeftesvoldoen. Die groter fokus op produktiwiteit in die landbou behoort die algemene verbruik van effektiewe boerderypraktyke, effektiewe kapitaalspandering, effektiewe implementering van navorsings- & ontwikkelingsresultate, produkdifferensiasie en handelsmerk-uitbouing in te sluit, asook die ontwikkeling van distribusiekanale met die vermoë om produkte koste effektief en tydig aan die mark te lewer. Daar bestaan geen enkele "super hoofweg" nie. Roetes is kompleks en nie volhoubaar nie, met ewigdurende veranderings. Vandag lewe en werk ons in 'n tegnologies-ontwikkelde samelewing. Vinnige vooruitgang in alle industrië is aan die orde van die dag. Om te oorleef en welvaart te skep moet Douglas Wynkelder die pad betree van voortdurende vordering en verbetering van strategië, om sodoende aan die visie van GWK Bpk te voldoen. DIE VISIE VAN GWK IS OM IN VOLHOUBARE LANDBOUBESIGHEID VAN UITNEMENDHEID TE WEES

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Tannins and colour components in red wine are important quality parameters. These factors can be manipulated in the vineyard by grape growing techniques or in the cellar by different winemaking practices. Grape seeds make a significant contribution to tannin concentration in wine when compared to those from the skins and pulp. Tannins contribute to the ageing potential, organoleptic properties and stabilisation of red wine colour. The colour of a red wine is also influenced by malolactic fermentation, the biological process that transforms malic acid into lactic acid which normally leads to an increase in pH. The subsequent change in pH alters the anthocyanin equilibrium, the primary colour components in red wine. Oxygen contributes to the polymerisation of anthocyanins over time to form more stable pigments that are less sensitive to pH fluctuations and sulphur dioxide bleaching. Limited research has been done on the use of oxygen after alcoholic fermentation and the impact it has on the phenolic composition of red wines. Similarly, only a few studies have examined the impact of either the addition or removal of seeds to the phenolic composition of a red wine in combination with oxygen addition. Additionally, little published data seems to exist on the effect of different pHs on red wine’s phenolic and colour development after oxygen addition. In our results we have shown that it is possible to stabilise wine colour by adding supplementary seeds before alcoholic fermentation. This led to an increase in colour intensity in certain cases. Some red wines produced without seeds had significantly lower colour intensities. This clearly suggests that catechins and proanthocyanidins are extracted from seeds and contribute to wine colour as they combine with other pigments to stabilise wine colour. Spectrophotometric and HPLC analyses have shown that the total phenolic content increased with seed concentration. However, we have observed that a wine may possibly become saturated with phenols when supplementary seeds are added. Anthocyanin concentrations often decreased when oxygen was added, while polymeric phenols and polymeric pigments sometimes increased. When applying different oxygen dosages to a red wine on commercial scale with microoxygenation, it was found that monomeric anthocyanins decreased as more oxygen was added and this decrease in anthocyanins led to the formation of stable polymeric pigments. This was reflected in the significant increase in colour intensity for the wines receiving oxygen. Small differences were detected in the total phenol and tannin concentration for the control and oxygenated wines. However, some of these phenolic and colour differences disappeared during subsequent ageing of the wine. When making wines of different initial pHs, we observed that the colour density decreased as the pH increased. The application of oxygen reduced the decrease in colour during MLF, especially at a lower pH. The addition of oxygen did not result in significantly different polymeric pigment concentrations in the various pH treatments, although the results could have been different if the wines were aged. However, pH differences in the range between 3.4 and 4.0 did not significantly influence the phenol composition of the wines under our conditions. This study led to a better understanding on the effect of oxygen additions under different conditions on red wine's phenolic and colour composition. / AFRIKAANSE OPSOMMING: Tanniene en kleur komponente teenwoordig in rooiwyn is belangrike kwaliteit parameters. Hierdie faktore wat wynkwaliteit beïnvloed kan gemanipuleer word in die wingerd deur verskeie verbouingstegnieke toe te pas en in die kelder deur die toepassing van verskillende wynbereidingsmetodes. Die bydrae van sade tot die finale tannienkonsentrasie in rooiwyn is groot in vergelyking met dié van die pulp en doppe. Tanniene dra by tot die verouderingspotensiaal, organoleptiese eienskappe en die stabilisasie van die wynkleur. Die kleur van ‘n rooi wyn word ook beïnvloed deur appelmelksuurgisting (AMG), die biologiese proses wat appelsuur omskakel na melksuur en ‘n gevolglike toename in die pH van die wyn veroorsaak. Hierdie verandering in die pH van die wyn beïnvloed die antosianien ewewig, die primêre kleur komponente teenwoordig in rooiwyn. Suurstof dra by tot die polimerisasie van antosianiene oor tyd om meer stabiele kleur pigmente te vorm met ‘n hoër kleurintensiteit wat minder sensitief is teenoor pH veranderinge en die bleikingseffek van swaweldioksied. Beperkte navorsing is gedoen op die gebruik van suurstof na alkoholiese gisting en die impak daarvan op die fenoliese samestelling van ‘n rooiwyn. Slegs ‘n paar studies het die invloed van die verwydering of byvoeging van sade in kombinasie met suurstoftoediening op ‘n rooiwyn se fenoliese samestelling ondersoek. Dit wil voorkom of beperkte gepubliseerde data beskikbaar is oor die effek wat verskillende pH’s het op rooi wyn se fenoliese en kleurontwikkeling na suurstof byvoeging. Ons resultate het aangedui dat dit wel moontlik is om in sekere gevalle die kleur van ‘n rooiwyn te stabiliseer deur addisionele sade by te voeg voor alkoholiese fermentasie. Hierdie byvoeging het ‘n toename in kleurintensiteit tot gevolg gehad. Sekere wyne wat gemaak is sonder sade het ‘n kenmerkend laer kleur intensiteit gehad. Hierdie bevinding is ‘n duidelike bewys dat katesjiene en prosianidiene geëkstraheer word vanuit die sade en bydra tot wynkleur deurdat hulle met ander pigmente verbind om die kleur sodoende te stabiliseer. Spektrofotometriese en hoë druk vloeistof chromatografie (HDVC) analises het gewys dat die totale fenoliese konsentrasie neem toe met ‘n toename in saad konsentrasie. Daar is egter waargeneem dat ‘n wyn moontlik versadig kan raak met fenole wanneer addisionele sade bygevoeg word. Antosianien konsentrasies het meestal afgeneem wanneer suurstof bygevoeg is, maar polimeriese fenole en polimeriese pigmente het partykeer toegeneem. Met die toediening van verskillende suurstof dosisse tot ‘n rooiwyn op kommersiële skaal het ons bevind dat monomeriese antosianiene afneem wanneer meer suurstof bygevoeg word. Hierdie afname in antosianiene het egter gelei tot die vorming van stabiele polimeriese pigmente. Dié bevinding was gereflekteer in die toename in kleurintensiteit van wyne wat met suurstof behandel is. Klein verskille was waargeneem vir die totale fenol en tannien konsentrasies tussen die kontrole en wyne behandel met suurstof. Sekere van hierdie fenoliese kleur verskille het egter afgeneem tydens die daaropvolgende veroudering van die wyne. Wyne wat gemaak is met verskillende aanvanklike pH’s se kleurintensiteit neem af soos die pH toeneem. Die toediening van suurstof het die kleurverlies tydens AMG verminder, veral by ‘n laer pH. Die toediening van suurstof het nie verskillende polimeriese pigment konsentrasies by verskillende pH’s veroorsaak nie, maar ‘n verskil kon moontlik waargeneem word indien die wyne verouder was. pH verskille tussen 3.4 en 4.0 het egter nie die fenoliese samestelling van die wyne onder ons omstandighede beduidend beïnvloed nie. Hierdie studie het gelei tot meer kennis oor die effek van suurstoftoedienings onder verskillende kondisies op rooiwyn se fenoliese en kleursamestelling.

Thesis (MSc)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: The polysaccharides that are present in wine originate from the grapes, the fungi that grow on the grapes and from other microorganisms that come into contact with the must during winemaking. The grape-derived polysaccharides of most concern in winemaking are pectin, glucan and xylan that can be enzymatically degraded by pectinases, glucanases and xylanases, respectively. These are the main structural polysaccharides of the cell wall of the grape cell. Degradation of the cell walls will result in the separation and rupture of the grape cells, and cell wall-bound compounds will be released into the must. Treating the must with pectinase and macerating enzyme preparations can result in an increase in free-flow juice, an improvement in must clarification and filtration, and an increased extraction of phenols and tannins. The tannins that are extracted polymerise with anthocyanins in red wine during ageing, resulting in increased colour intensity and stability. Wine aroma is also influenced by enzyme treatment. The degradation of the cell wall contributes to the release of glycosidically-bound terpene or alcohol precursors from the berries. The hydrolysis of these precursors during fermentation can result in an improvement in aroma. It can thus be seen that it is possible to improve wine quality and processing by supplementing the endogenous enzymes that are present in the fermentation with commercial enzyme preparations. Commercial enzymes are typically crude fungal preparations. The majority of commercial pectinase and glucanase preparations are derived from Aspergillus and Trichoderma, respectively. Since the endogenous polysaccharase activity of Saccharomyces cerevisiae is very limited, the heterologous expression of specific polysaccharase genes in an industrial yeast strain can improve the winemaking process, resulting in a higher quality wine without the addition of expensive commercial enzyme preparations. Since only the desired enzymes are secreted by the recombinant strain, there will be no undesired sideactivities, which can be detrimental to wine quality. Several pectinase-, glucanaseand xylanase-encoding genes, cloned from a variety of organisms, have been expressed successfully in laboratory strains of S. cerevisiae. Attempts have also been made to construct industrial wine yeast strains that express these polysaccharase genes and secrete the encoded enzymes. Fermentation with some of these strains resulted in a decrease in total phenolics and turbidity, an increase in juice extraction, and alterations in the colour and aromatic profile of the resulting wines. In this study, four polysaccharide-degrading, recombinant wine yeast strains were constructed. The endo-β-1,4-xylanase gene, XYN2, and the endo-β-1,4-glucanase gene, end1, were previously cloned from the soft rot fungus Trichoderma reesei and the rumen bacterium Butyrivibrio fibrisolvens, respectively. These genes were subcloned into different expression cassettes which were used to construct the four integration plasmids. The recombinant plasmids contained the following gene cassettes: TEF1P-XYN2-ADH2T (plasmid pDLG29) ADH1P- MFα1S -end1-TRP5T (plasmid pDLG30) ADH1P-MFα1S-end1-TRP5T and ADH2P-XYN2-ADH2T (plasmid pDLG33), ADH1P-MFα1S-end1-TRP5T and YG100PXYN2- ADH2T (plasmid pDLG39). These four plasmids were then separately integrated into the ILV2 locus of the commercial wine yeast strain S. cerevisiae VIN13. Wine was made with the four strains constructed in this study, a pectolytic strain, VIN13[pPPK], a glucanase- and xylanase-secreting strain, VIN13[pEX], an untransformed VIN13 strain, and an untransformed strain with the addition of the commercial enzyme preparation Rapidase EX Colour. Microvinification experiments were carried out on Pinot noir, Ruby Cabernet and Muscat d’Alexandria wines. Fermentation with the polysaccharide-degrading strains resulted in significant improvements in juice extraction, colour intensity and stability, and in alterations in the aromatic profiles of the wines produced. Subject to the approval by the regulatory authorities and eventual consumer acceptance of the use of genetically modified organisms (GMOs) in fermented foods and beverages, it might be required that the GM status of the yeast that is used appears on the label. Currently, there is no robust technique available with which the use of GM yeast can be revealed in a finished wine because the yeast cells and their DNA are removed from or denatured in the wine during filtration and processing. One way with which the undeclared use of a GM yeast in winemaking could be exposed would be to compare the chemical profile of a suspect wine with that of non-GM wine. In order to explore this concept further, a secondary aim of this study was to investigate whether Fourier Transformation Infra Red (FT-IR) spectroscopy coupled with multivariate data analysis could distinguish between wines fermented with transgenic and non-transgenic yeast strains, or between wines fermented with different transgenic strains. The results showed that this method could be used to classify wines fermented with different yeast strains if fermentation with the strain resulted in a unique chemical profile in the resulting wine. This was a preliminary study and these findings were summarised as an addendum to the thesis. / AFRIKAANSE OPSOMMING: Die polisakkariede wat in wyn teenwoordig is, is afkomstig van die druiwe, die swamme wat op die druiwe groei en vanaf ander mikroörganismes wat tydens die wynmaakproses met die mos in aanraking kom. Die belangrikste druifpolisakkariede in wynbereiding is pektien, glukaan en xilaan, wat onderskeidelik deur pektinases, glukanases en xilanases afgebreek kan word. Hierdie is die vernaamste strukturele polisakkariede van ‘n druifsel se selwand. Die afbreking van die selwande veroorsaak dat die druifselle skei en skeur, met die gevolg dat die selwandgebonde verbindings in die mos vrygelaat word. Die behandeling van die mos met pektinase en versappingsensiempreparate kan tot ʼn toename in vry-afloopsap lei, sowel as ʼn verbetering in mosverheldering en -filtrasie en ʼn verhoogde ekstraksie van fenole en tanniene. Die tanniene wat geëkstraheer word, polimeriseer in rooiwyn tydens veroudering, en dit lei tot verhoogde kleurintensiteit en -stabiliteit. Wynaroma word ook deur ensiembehandeling beïnvloed. Die afbreking van die druifselwand dra by tot die vrylating van glikosidiesgebonde terpeen- en alkoholvoorlopers uit die korrels. Die hidrolise van hierdie voorlopers tydens gisting kan lei tot ʼn verbetering van die aroma. Dit is dus duidelik dat dit moontlik is om wynkwaliteit en wynbereiding te verbeter deur die endogene ensieme wat in die gisting teenwoordig is met kommersiële ensiempreparate te supplementeer. Kommersiële ensiempreparate is tipies ongesuiwerde swampreparate. Die meerderheid kommersiële pektinase- en glukanasepreparate word onderskeidelik vanaf Aspergillus en Trichoderma verkry. Aangesien die endogene polisakkaraseaktiwiteit van Saccharomyces cerevisiae baie beperk is, kan die heteroloë uitdrukking van spesifieke polisakkarase-gene in ʼn industriële gisras die wynbereidingsproses verbeter en lei tot ʼn hoër kwaliteit wyn sonder die byvoeging van duur kommersiële ensiempreparate. Omdat die verkose ensieme deur die rekombinante ras uitgeskei word, sal daar geen ongewenste newe-effekte teenwoordig wees wat ʼn nadelige effek op wynkwaliteit kan hê nie. Verskeie mikrobiese gene wat vir pektinases, glukanases en xilanases kodeer, is reeds voorheen uit ‘n wye verskeidenheid van organismes gekloneer en suksesvol in laboratoriumrasse van S. cerevisiae uitgedruk. Pogings is ook aangewend om industriële wyngisrasse te konstrueer wat hierdie polisakkarasegene uitdruk en hul enkodeerde ensieme uitskei. Gisting met sommige van hierdie rekombinante gisrasse het gelei tot ʼn afname in totale fenoliese verbindings en troebelheid, ʼn verhoging in sapekstraksie, en veranderings in die kleur en aromatiese profiel van die gevolglike wyne. In hierdie studie is vier polisakkaried-afbrekende, rekombinante wyngisrasse gekonstrueer. Die endo-β-1,4-xilanasegeen, XYN2, en die endo-β-1,4- glukanasegeen, end1, is voorheen reeds onderskeidelik vanaf die sagte vrotswam, Trichoderma reesei, en die rumenbakterium, Butyrivibrio fibrisolvens, gekloneer. Hierdie gene is in vier integrasieplasmiede in verskillende ekspressiekassette gesubkloneer. Die plasmiede het die volgende geenkassette bevat: TEF1P-XYN2- ADH2T (plasmied pDLG29) ADH1P- MFα1S -end1-TRP5T (plasmied pDLG30) ADH1PMFα1S- end1-TRP5T and ADH2P-XYN2-ADH2T (plasmied pDLG33), ADH1P-MFα1S end1-TRP5T and YG100P-XYN2-ADH2T (plasmied pDLG39). Hierdie vier plasmiede is toe afsonderlik in die ILV2-lokus van die kommersiële wyngisras, S. cerevisiae VIN 13, geïntegreer. Wyn is met hierdie vier gekonstrueerde gisrasse gemaak, die pektolitiese gisras, VIN13[pPPK], die glukanase- en xilanase-afskeidende gisras, VIN13[pEX], die ongetransformeerde VIN13-ras, en met ʼn ongetransformeerde VIN13 gis waarby die kommersiële ensiempreparaat, Rapidase EX Colour, bygevoeg is. Mikro-wynbereidingseksperimente is op Pinot noir-, Ruby Cabernet- en Muscat D’Alexandria wyne uitgevoer. Gisting met die polisakkaried-afbrekende gisrasse het gelei tot ʼn noemenswaardige verbetering in sapekstraksie, kleurintensiteit en kleurstabiliteit, asook in veranderinge in die aromatiese profiele van die geproduseerde wyne. Indien die gebruik van geneties gemodifiseerde organismes (GMOs) in gefermenteerde voedsel en drank deur die reguleringsowerhede goedgekeur en uiteindelik deur die verbruiker aanvaar sou word, sou dit vereis kon word dat die GMstatus van die wyngisgis op die etiket van die wynbottel aangebring word. Verpligte etikettering van GM-wyn sal metodes vereis waarmee die ‘nalentskap’ van GMgisselle in die finale produk geïdentifiseer en gemoniteer kan word. Tans is daar geen robuuste tegnieke beskikbaar waarmee die gebruik van GM-giste openbaar kan word nie, aangesien die gisselle en hul DNA tydens filtrasie en prosessering verwyder word. Een wyse waarop die onverklaarde gebruik van ‘n GM-gis in wynbereiding blootgestel sou kno word, is om die chemiese profiel van die verdagte wyn met dié van ‘n nie-GM-wyn te vergelyk. Ten einde hierdie konsep verder te ondersoek was ‘n sekondêre doelwit van hierdie studie om te bepaal of FT-IR (Fourier-transformasie-infrarooi) spektroskopie tesame met meervariante dataanalise gebruik kan word om te onderskei tussen wyne wat met transgeniese en nietransgeniese gisrasse gegis is, of tussen wyne wat met verskillende transgeniese rasse gegis is. Die resultate het aangedui dat hierdie metode gebruik kan word om wyne wat met verskillende gisrasse gegis is, te klassifiseer indien die betrokke gisras ʼn unieke chemiese profiel in die uiteindelike wyn veroorsaak het. Dit was egter ʼn voorlopige ondersoek en is as ʼn byvoegsel tot die tesis geskryf.

Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Glutathione (GSH), a tripeptide consisting of glutamate, cysteine and glycine, is the most ubiquitous non‐protein intracellular thiol in a large variety of organisms, including plants, animals and fungi. The thiol moiety of the cysteine residue confers unique redox and nucleophilic properties. In plant cells, GSH fulfils an indispensible role in the antioxidant system, sulphur metabolism and detoxification of xenobiotics. Upon grape crushing, GSH is extracted into the juice where it exerts several protective effects during the vinification process. In must, it reacts with oxidized phenolic compounds to form the colourless grape reaction product (GRP) which limits must browning to a certain extent. During wine ageing, GSH impedes the decrease of important aroma compounds, including certain esters, terpenes and volatile thiols, while at the same time preventing the development of atypical ageing off‐flavours. GSH may also inhibit the yellowing of wine during ageing. It is thus evident that elevated GSH levels in wine, in particular white wine which is more sensitive to oxidation, may be highly valuable for wine quality. The reductive crushing and pressing of white grape varieties, which limits oxidation and the consequent incorporation of GSH into GRP, promotes higher GSH levels in the juice. The reductive handling of juice also limits the formation of oxidized glutathione (GSSG). However, during alcoholic fermentation and maturation, levels generally decrease as a result of assimilation by the yeast Saccharomyces cerevisiae, and inevitable oxidation that takes place during the vinification process. The principal focus of this study was to gain a better understanding of the fate of glutathione during alcoholic fermentation and to establish whether certain oenological applications could result in elevated wine GSH levels. The application studied, included choice of yeast strain, extended lees contact, nitrogen supplementation and supplementation with glutathione enriched inactive dry yeast preparations (GHS‐IDYs). In addition, the need for a rapid analytical method for the simultaneous quantification of both GSH and GSSG in must and wine which does not involve derivatization or require extensive sample preparation, led to the development of a novel UPLC‐MS/MS method. The method was also employed to determine intracellular GSH and GSSG contents of the yeast S. cerevisiae and was studied for the first time in winemaking conditions. It was shown that the GSH levels fluctuated during alcoholic fermentation, suggesting the uptake and release by yeast. At the end of alcoholic fermentation, levels were generally lower than those initially present in grape juice, but in some cases, concentration increases were also observed. This finding indicates that, in some cases, endogenously‐produced GSH may be secreted into must during alcoholic fermentation, contributing to higher GSH levels in wine. Albeit small, significant differences in GSH content could be seen in wines fermented with different yeast strains, implying that yeast strain may to a certain extent influence wine GSH levels. While the effects of lees ageing and nitrogen supplementation seem to be insignificant in contributing to higher GSH levels in wine, the supplementation of must with GSH‐IDYs could result in increased wine GSH levels, provided the supplementation is done early during fermentation. This study has broadened our knowledge of several oenological factors, influencing GSH levels in wine and provided a new baseline for future research studies. / AFRIKAANSE OPSOMMING: Glutatioon (GSH), ’n tripeptied bestaande uit glutamaat, sisteïen en glisien, is die mees algemene nieproteïenagtige intrasellulêre tiool in ‘n wye verskeidenheid organismes, insluitende plante, diere en fungi. Die tioolfunksiegedeelte van die sisteïenresidu verleen unieke redoks‐ en nukleofiliese eienskappe. GSH vervul ‘n onmisbare rol in die antioksidantsisteem, swaelmetabolisme en die ontgiftiging van xenobiotika in plantselle. Tydens die maal van druiwe word glutatioon in die sap geëkstraeer waar dit verskeie beskermende effekte tydens die wynbereidingsproses uitoefen. GSH reageer met geöksideerde fenoliese verbindings om die kleurlose druifreaksieproduk (DRP) te vorm wat die verbruining van mos in ‘n sekere mate beperk. GSH verminder ook die afname van belangrike aromaverbindings tydens wynveroudering, insluitende sekere esters, terpene en vlugtige tiole, terwyl dit terselfdertyd die vorming van atipiese verouderingswangeure belemmer. So ook kan GSH die vergeling van wyn tydens veroudering inhibeer. Dit is dus voor die hand liggend that verhoogde GSH‐vlakke in wyn, in die besonder witwyn, wat meer oksidasie‐sensitief is, van waarde kan wees vir wynkwaliteit. Die reduktiewe maal en pers van witdruifvariëteite wat oksidasie en gevolglike inkorporasie van GSH in DRP beperk, bevorder hoër GSH‐vlakke in sap. So ook beperk die reduktiewe behandeling van sap die vorming van geoksideerde glutatioon (GSSG). Gedurende alkoholiese gisting en veroudering neem GSH‐vlakke egter af as gevolg van assimilasie deur die gis, Saccaromyces cerevisiae, asook onvermydelike oksidasie wat gedurende die wynbereidingsproses plaasvind. Die hooffokus van die studie was om ‘n beter begrip van die lot van glutatioon tydens alkoholiese gisting te verkry en om vas te stel of sekere wynkundige praktyke verhoogde GSH‐vlakke in wyn tot gevolg kan hê. Die studie het gisraskeuse, verlengde gismoerkontak, stikstofaanvulling en aanvulling met glutatioon‐verrykte, onaktiewe droëgis ingesluit. Daarbenewens het die behoefte aan ‘n vinnige analitiese metode vir die gelyktydige kwantifisering van sowel GSH as GSSG in mos en wyn wat nie derivatisering of uitgebreide monstervoorbereiding vereis nie, gelei tot die ontwikkeling van ‘n nuwe UPLC‐MS/MS metode. Hierdie metode is ook gebruik om die intrasellulêre GSH‐ en GSSG‐inhoud van die gis S. cerevisiae te bepaal wat vir die eerste keer in wynbereiding bestudeer is. Daar is bewys dat GSH‐vlakke tydens alkolholiese gisting fluktueer, wat dui op die opname en vrystelling daarvan deur die gis. Die vlakke aan die einde van alkoholiese gisting was oor die algemeen laer as vlakke aanvanklik teenwoordig in die sap. In sommige gevalle is konsentrasietoenames egter ook waargeneem. Hierdie bevinding dui daarop dat intrasellulêr‐vervaardige GSH, in sommige gevalle, in die mos uitgeskei kan word, wat tot hoër GSH‐vlakke in wyn lei. Klein, dog beduidende verskille in GSHinhoud is waargeneem in wyne wat met verskillende gisrasse berei is, wat daarop dui dat gisras in ‘n sekere mate die GSH‐vlakke in wyn kan beïnvloed. Alhoewel die effek van gismoerveroudering en stikstofaanvulling onbeduidend is, kan die aanvulling van mos met glutatioon‐verrykte, onaktiewe droëgis tot verhoogde GSH‐vlakke in wyn lei, mits die aanvulling vroeg tydens alkoholiese gisting gedoen word. Hierdie studie verbreed ons kennis van verskeie wynkundige praktyke wat GSH‐vlakke in wyn beïnvloed en vorm ‘n nuwe basis vir toekomstige navorsingstudies.

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2009. / South Africa is the eighth largest wine producing country in the world and face stiff competition on the world market. Cabernet Sauvignon is the most planted red cultivar in the world as well as in South Africa and can be seen as the wine by which countries are judged. The aim of this study was to investigate suitable, practical maturity parameters or combinations thereof to determine the optimal time to harvest Cabernet Sauvignon grapes under South African conditions. The following parameters were investigated during this study: seed lignification, maturity indexes, anthocyanin concentration per berry, sensory criteria (grape skins tasting and wine) and phenolic content. Berry development in four Cabernet Sauvignon vineyards in different South African winegrowing areas were investigated over the 2003, 2004 and 2005 seasons. The first parameter to be investigated was seed lignification percentages. Seasonal differences at commercial harvest were observed with values of 2004 varying between 73% and 91% compared to 59% and 80% for the 2003 and 2005 seasons but commercial harvest was two weeks later during the 2004 season. During this study it was found that seeds never reached 100% lignification for Cabernet Sauvignon as was found in previous work to indicate grape maturity. The development of anthocyanins also peaked well before the maximum seed lignification was reached. It therefore appears that seed lignification is not suitable for the determination of grape maturity for Cabernet Sauvignon grapes under South African conditions. The second parameter to be investigated was maturity indexes (Balling / Titratable Acidity (TA), Balling × pH, Balling × pH2). The best wine values were used to determine the optimal maturity index values. Morgenster was the only vineyard to consistently give values that corresponded to previously reported data (index values). Anhöhe and Plaisir de Merle reported higher maturity values than that reported in literature and seasonal variation was observed. Maturity index values for the best wines varied between 88 and 101 (Balling × pH) for Anhöhe during 2003 and 2005 seasons, but increased too between 97 and 107 (Balling × pH) for 2004. The maturity index values were found to be vineyard and season dependant, with warmer areas reaching higher values. From this study it appears that maturity index values as a singular maturity parameter does not give a good indication of berry maturity in all seasons or vineyards. Thirdly, the berry anthocyanin concentration (mg / berry and mg / g berry) were investigated and comparable trends were found between the four vineyards. However vineyards in warmer, drier regions (Anhöhe) tended to have higher anthocyanin concentrations per gram berry. The more vigorous vineyard of Morgenster consistently exhibited a higher anthocyanin concentration per berry. This can be explained by the ratio of skin to pulp between small berries (Anhöhe, 0.95 g - 2004) and larger berries South Africa is the eighth largest wine producing country in the world and face stiff competition on the world market. Cabernet Sauvignon is the most planted red cultivar in the world as well as in South Africa and can be seen as the wine by which countries are judged. The aim of this study was to investigate suitable, practical maturity parameters or combinations thereof to determine the optimal time to harvest Cabernet Sauvignon grapes under South African conditions. The following parameters were investigated during this study: seed lignification, maturity indexes, anthocyanin concentration per berry, sensory criteria (grape skins tasting and wine) and phenolic content. Berry development in four Cabernet Sauvignon vineyards in different South African winegrowing areas were investigated over the 2003, 2004 and 2005 seasons. The first parameter to be investigated was seed lignification percentages. Seasonal differences at commercial harvest were observed with values of 2004 varying between 73% and 91% compared to 59% and 80% for the 2003 and 2005 seasons but commercial harvest was two weeks later during the 2004 season. During this study it was found that seeds never reached 100% lignification for Cabernet Sauvignon as was found in previous work to indicate grape maturity. The development of anthocyanins also peaked well before the maximum seed lignification was reached. It therefore appears that seed lignification is not suitable for the determination of grape maturity for Cabernet Sauvignon grapes under South African conditions. The second parameter to be investigated was maturity indexes (Balling / Titratable Acidity (TA), Balling × pH, Balling × pH2). The best wine values were used to determine the optimal maturity index values. Morgenster was the only vineyard to consistently give values that corresponded to previously reported data (index values). Anhöhe and Plaisir de Merle reported higher maturity values than that reported in literature and seasonal variation was observed. Maturity index values for the best wines varied between 88 and 101 (Balling × pH) for Anhöhe during 2003 and 2005 seasons, but increased too between 97 and 107 (Balling × pH) for 2004. The maturity index values were found to be vineyard and season dependant, with warmer areas reaching higher values. From this study it appears that maturity index values as a singular maturity parameter does not give a good indication of berry maturity in all seasons or vineyards. Thirdly, the berry anthocyanin concentration (mg / berry and mg / g berry) were investigated and comparable trends were found between the four vineyards. However vineyards in warmer, drier regions (Anhöhe) tended to have higher anthocyanin concentrations per gram berry. The more vigorous vineyard of Morgenster consistently exhibited a higher anthocyanin concentration per berry. This can be explained by the ratio of skin to pulp between small berries (Anhöhe, 0.95 g - 2004) and larger berries (Morgenster, 1.82 g – 2004). Wine colour density (A420+A520) followed the same trend as the anthocyanin concentrations of the homogenate. Grape skins (G) were used to make an artificial wine that was evaluated by an expert panel to determine the development of the grapes. Wines (W) made from sampled batches were also evaluated by an expert panel for: colour intensity, vegetative, red berry, black berry with spice, acidity, astringency and general quality. Vegetative aromas and acidity decreased and red and black berry with spice increased during ripening for both berries and wine. Colour intensity also increased, corresponding to an increase in perceived general quality score. Correlations between general quality of both the grape skins tasting and wines were investigated. Balling showed a strong correlation with general quality of the grape skins tasting (r = 0.76; p = 0.00) but not as strongly with subsequent wines (r = 0.57; p = 0.00). Anthocyanin concentration (mg / g berry) of the berries (r = 0.36; p = 0.00), perceived colour intensity of grapes (r = 0.69; p = 0.00) and wine (r = 0.84; p = 0.00) correlated with general wine quality. The tasting panel identified wines that were statically better than the rest for each season and vineyard. Maximum berry anthocyanin concentration coincided with wines rated as the best by the tasting panel. More than one wine was identified during the maximum anthocyanin peak that did not differ statistically from the best wine. It appears from this study that a window period exists at the maximum anthocyanin peak, where wines of comparable quality, but different style, can be produced. Principal component analysis (PCA) was used to determine the least number of suitable parameters that could distinguish between unripe and ripe grapes in order to establish a grape maturity model. These differences were successfully described by Balling, TA, pH, potassium (K+), tartaric and malic acid. Anthocyanin concentration could further distinguish between ripe and overripe grapes in the model. From these parameters the minimum and maximum values were used to construct a universal ripeness model containing data from all four vineyards. Variation between the four vineyards caused too much overlapping in the universal model data as the vineyards were situated in different climatic regions according to the Winkler temperature model. On a per vineyard basis this did not occur to the same extend. The best rated Cabernet Sauvignon wines correlated strongly with soluble solid content; colour and quality perceptions of grapes, but large seasonal differences resulted in larger grape compositional variances than that of the individual vineyards in the different climatic zones. This illustrated the difficulty of pinpointing a specific parameter to indicate optimal ripeness. From this study it is clear that a universal maturity model for Cabernet Sauvignon berries is not attainable at present, but individual vineyard models shows the most potential. A preliminary study into the differences of the phenolic composition was done using reverse phase high performance liquid chromatography (RP-HPLC) on the homogenate and wine. Malvidin-3-glucoside and total anthocyanins followed comparable trends to that found for the Iland method. Strong correlations (r > 0.9) were found between the malvidin- 3-glucoside and malvidin-3-glucoside-acetate and p-coumarate; this was also true for the total anthocyanins in both homogenate and wine. Wines identified by a tasting panel to be the best quality, corresponded with the maximum anthocyanin concentration (mg / L) peak in the homogenate. Dense canopies at the Morgenster vineyard over the three seasons lead to lower total anthocyanin and quercetin-3-glucuronide concentrations compared to the Anhöhe and Plaisir de Merle vineyards. The shading of bunches by the dense canopy most likely contributed to this. Catechin, epicatechin, proanthocyanidin and polymeric phenol concentrations decreased significantly from veraison until harvest. Seasonal differences were noted in the four vineyards. No correlations could be found between the general wine quality and the phenolic compounds, but a weak trend was observed for total anthocyanins in the homogenate. A trend was found with the total flavan-3-ol to anthocyanin ratio determined by RP-HPLC analysis of the grape homogenates (r = 0.40, p = 0.00). This ratio varied between 1 and 3 for the wines rated as being the best quality. Phenols by themselves do not give a clear indication of optimal harvest time. From this study it appears that no single parameter could consistently indicate optimal ripeness over the seasons or per vineyard, but the maximum berry colour (anthocyanin concentration) did give an indication of optimal harvesting time. It is clear that a combination of parameters could predict the optimal time more precisely as with the above mentioned model but more research is needed to this end.

Thesis (MSc (Wine Biotechnology))--University of Stellenbosch, 2006. / Resveratrol is a phytoalexin that is produced in the leaves and skins of grape berries in response to biotic and abiotic factors. Substitution and polymerisation of resveratrol units produce an array of compounds which form part of the active disease defence mechanism in grapevine. Wine is one of the major sources of resveratrol in the human diet. Resveratrol is one of the phenolic compounds present in wine that mediates protective effects on human health. It has been shown to prevent the development of cardiovascular disease, cancer and pathogenesis related to inflammation. Red wines contain higher levels of resveratrol than white wines owing to extended maceration times during fermentation on the skins. During white wine vinification skin contact is limited as skins are removed prior to fermentation. Thus, the extraction of resveratrol into white wines is minimal. The principal focus of our research is the development of a wine yeast strain capable of resveratrol production during grape must fermentation. It is proposed that red and white wines produced with such a resveratrol-producing yeast will contain elevated levels of resveratrol, and that added health benefits may be derived from their consumption. Initial work done in our laboratory established that expressing multiple copies of the genes encoding coenzyme A ligase (4CL216) and resveratrol synthase (vst1) in laboratory yeast enabled the yeast to produce resveratrol, conditional to the supplementation of the growth medium with p-coumaric acid. This study focused on the optimisation of resveratrol production in Saccharomyces cerevisiae. It involved the integration and constitutive expression of 4CL216 from hybrid poplar and vst1 from grapevine. Integration and expression of these genes in three laboratory strains was confirmed by Southern and Northern blot analyses. The evaluation of resveratrol production by yeast required the initial optimisation of the analytical techniques. We optimised the method for sample preparation from the intracellular fraction of yeast and devised a procedure for the assay of the extracellular fractions. The LCMSMS method was further developed to encompass detection and quantification of other compounds related to resveratrol production in yeast. Comparison of resveratrol production in three different yeast genetic backgrounds indicated that the onset of production and the resveratrol yield is yeast strain dependent. Precursor feeding studies indicated that p-coumaric acid availability was a factor limiting maximal resveratrol production. Early indications were obtained that endogenously-produced resveratrol may have an impact on yeast viability during extended culture periods. This study has broadened our understanding of the resveratrol production dynamics in S. cerevisiae and provided important indications as to where further optimisation would be beneficial in order to optimally engineer a wine yeast for maximal resveratrol production.

Thesis (MSc (Wine Biotechnology))--Stellenbosch University, 2008. / In the wine industry, the importance of selecting an appropriate yeast strain, generally of the species Saccharomyces cerevisiae, to ensure reliable fermentation and to achieve a desired level of quality has been well established. As a consequence, the demand for new starter cultures with improved or new oenological characteristics is increasing. Appropriately selected starter cultures can reduce the occurrence of stuck fermentations, impart specific aroma profiles and reduce the development of offflavours. Using standard breeding and selection procedures, several wine yeast strains that would be less likely than currently existing strains to experience stuck fermentation have previously been developed at the Institute for Wine Biotechnology. The target of these projects had been to develop strains with improved nitrogen efficiency [defined as the amount of fermented hexoses for a given amount of free amino nitrogen (FAN)], improved fructose utilization and ethanol tolerance. These three parameters are known contributors to stuck fermentation. Two of the strains that had been isolated in these projects, strain 116 for nitrogen efficiency and strain 38-1 for efficient fructose utilization, were chosen as parental strains for the current study. The aim was to further improve and possibly combine these traits in yeast strains by using hybridization followed by various enrichment and directed evolution procedures in a continuous fermentation setup. The strategy was to sequentially subject the population of mass-mated hybrids to a number of selective environments for a large number of generations. The yeasts were subjected to a high fructose/glucose ratio for 12 generations, followed by selection in an environment with a limited supply of nitrogen for 54 generations and finally to high ethanol stress. After each round of enrichment, individual strains were analysed to assess the results. For the hybrid strains selected after enrichment in a medium with a high fructose/glucose ratio, no general improvement could be discerned. However, one of the hybrids, hybrid strain 331, fermented fructose better than the parental strains and other hybrid strains. These results may suggest that the selection pressure was not applied for a sufficient number of generations and may not have been sufficiently strong. In addition, the parental strain may already performing at a rate that may render further improvement more difficult in this genetic background. The next aim of this study was to enhance fermentation performance of wine yeast hybrid strains in low nitrogen and high sugar conditions. Several hybrid strains 331, RR03 and 05R generated in this study showed improvement in efficiency of nitrogen utilization when compared to the parental strains, indicating a successful selection strategy. Several strains also showed higher ethanol tolerance, and some strains possessed] combinations of the traits to be improved. Future research will evaluate these hybrids regarding the production of aromatic compounds and of the sensory profile produced. Such strains would help the wine industry to control the occurrence of stuck fermentations and to produce quality wines.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2006. / The assessments of grape and wine quality are complex issues and the wine industry needs more objective analysis of grape and wine quality. The standard quality assessment protocol for grading grapes at most wine cellars in South Africa is based on viticultural practices and the determination of chemical parameters such as ºBrix, pH and titratable acidity (TA). Grape juice indices calculated by formulae such as ºBrix/pH, TA/pH, ºBrix/TA, ºBrix x (pH)2 have been used in the past but these approaches have had limited success. It was shown that the total anthocyanins and total phenolics of red grapes correlate with wine quality and provide additional objective measures of grape quality. Most methods for the quantification of total anthocyanins and total phenolics are complex and time consuming and therefore not easily implemented in the routine laboratory environment. Fourier transform infrared spectroscopy (FT-IR) is widely used in South African laboratories for the routine quantification of wine and grape parameters but the commercial calibration models supplied for the quantification of grape total anthocyanins and phenolics are not satisfactory. The focus of this study was to develop new FT-IR calibration models for the quantification of total anthocyanins and phenolics of grapes and to use the generated data during a preliminary evaluation of the implementation of these parameters as part of the grape quality control protocol at a commercial winery in South Africa. The potential of Fourier transform infrared spectroscopy (FT-IR) for the rapid quantification of total anthocyanins and total phenolics in red grapes was investigated and evaluated for prediction accuracy with independent validation sets. The design of calibration sets aimed at capturing most of the variation due to vintage (2004 and 2005), cultivar (Cabernet Sauvignon, Merlot, Pinotage, and Shiraz) and sugar concentration. Best prediction accuracies were obtained for calibration sets using grapes from a single vintage or cultivar or approximately the same sugar concentration. The highest prediction accuracies were obtained for total anthocyanins calibration sets of grapes with sugar concentrations ≥ 23.5ºBrix (SEP = 0.13 mg/g; R2 validation set = 0.77) and for total phenolics calibration sets of grapes with sugar concentrations < 23.5ºBrix (SEP = 0.13 OD280/g; R2 validation set = 0.74). Strong correlations were found between the spectral data and the total anthocyanins (SEP = 0.12 mg/g; R2 validation set = 0.84) and total phenolics concentration data (SEP = 0.10 OD280/g; R2 validation set = 0.76) for 2005 Merlot calibration sets indicating that the FT-IR spectra captured most of the variation. Overall the RPD (ratio of the standard deviation of the reference data to the standard error of prediction) values of all calibration models were below 3 indicating that calibration models are fit for screening purposes. Spectroscopic absorbance at 280 nm is not specific enough for the quantification of total phenolics and the use of an alternative reference method such as high performance liquid chromatography (HPLC) will be considered in the future. Principal component analysis (PCA) revealed that the major sources of variation in the FT-IR spectra of grapes could be ascribed to vintage and grape sugar concentration and this had an effect on the accuracy of the analytical data generated when using FT-IR spectroscopy. This report is the first to our knowledge where FT-IR has been used for the quantification of total anthocyanins and phenolics of grapes. The evaluation of the reference laboratory protocol for the quantification of total anthocyanins and total phenolics in grapes were evaluated in Chapter 4 and emphasized the importance of meticulous laboratory practices to obtain reliable reference data for calibration purposes. This large scale investigation of the total anthocyanins and phenolics concentrations in grapes is the first of its kind in South Africa and a quantitative database containing analytical data of the anthocyanins and total phenolics concentrations of 692 grape samples representing a wide range of grape maturities of Vitis vinifera cultivars Cabernet Sauvignon, Merlot, Pinotage and Shiraz from the 2004 and 2005 vintages was established based on the reference values. The data were used in a preliminary investigation of the implementation of total anthocyanins and total phenolics concentrations as part of grape quality classification at a commercial South African winery (Chapter 5). The results showed that the total anthocyanins and total phenolics concentration in grapes increased with increasing grape maturity (measured as sugar concentration). ANOVA and post-hoc analysis (Bonferroni testing) revealed significant differences between the total anthocyanins and total phenolics concentrations of the four Vitis vinifera cultivars investigated. Grapes harvested earlier in the season had significantly higher (p≤0.05) total anthocyanins and total phenolics concentrations than grapes harvested later in the season. This implies that grapes harvested earlier in the harvest season could produce wines with higher quality. Grapes from regularly irrigated vineyards had lower total anthocyanins and total phenolics concentrations compared to dryland vineyards. The current grape grading system in use at the industrial cellar did not correlate well with the total anthocyanins and total phenolics concentrations of grapes which highlighted the need for the inclusion of more objective measures during grape grading. The information captured in the database can be used as a starting point to establish profiles of the typical anthocyanins and total phenolics of South African grapes and data from more vintages should be included and continually updated. These findings highlight the important contribution of the results obtained in this preliminary study for the incorporation of total anthocyanins and phenolics concentrations as objective parameters of grape quality. Finally multivariate data analysis of the FT-IR spectra revealed important information regarding factors (both physical and chemical) that contribute to the variation of the spectra. The main variation between the 2004 and 2005 samples can probably be interpreted in terms of the water content of the samples.