Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: During wine fermentation, numerous grape must constituents serve as nutrients to wine yeast (Saccharomyces cerevisiae), which enable their growth and successful completion of alcoholic fermentation. Many of these nutritional factors, in particular nitrogen, also act as precursors for yeastderived flavour compounds such as higher alcohols, esters and volatile fatty acids. Yeast nitrogen metabolism thus plays a determining role in wine aroma and quality. Not only is the nitrogen source, concentration and supplementation timing important, but various environmental factors and the genetic constitution of the yeast strain used for fermentation will also contribute to fermentation outcomes. The main goal of this work was to explore the complex interactions between a number of contributing factors; namely nitrogen source, timing of addition, yeast strain and fermentation matrix. Broadly, this study assessed the impact of seven different nitrogen combinations, added either to the initial grape must or after the onset of fermentation, on fermentation performance and aroma compound production by nine commercial wine yeast strains. Fermentations were done in synthetic grape must, and validated for a subset of parameters in real grape must. The nitrogen treatments were designed according to the generally established order of preference of S. cerevisiae for individual amino acids as source of nitrogen under fermentative conditions, and the potential of certain amino acids to participate in metabolic pathways that produce specific aroma compounds. The results reveal that different nitrogen combinations can lead to unexpected aroma outcomes, depending strongly on the genetic background of individual yeast strains and the timing of nitrogen addition. Certain nitrogen treatments consistently resulted in significant increases or decreases in specific aroma compound concentrations in comparison to the treatment fermented on ammonium as only nitrogen source, for multiple yeast strains. These compounds were classified as nitrogen treatment dependent. Other aroma compounds were produced similarly for all nitrogen treatments and were designated as nitrogen treatment independent. The presence of specific amino acid groups (for example the branched-chain and aromatic amino acids) could be correlated to significantly altered production patterns of related (such as higher alcohols) or unrelated (diethyl succinate) aroma compounds relative to the other nitrogen treatments. Taken together, a number of interesting and novel hypotheses regarding the metabolic pathways involved could be derived from the data. Ultimately, this initial assessment of interactive effects during fermentation will contribute to practical guidelines for winemakers to allow matching grape must constituents (such as nutrients) with the intrinsic aroma production capabilities of specific yeast strains in order to modulate wine aroma, style and quality. / AFRIKAANSE OPSOMMING: Tydens wynfermentasie dien talle druiwemosbestanddele as voedingstowwe vir wyngis (Saccharomyces cerevisiae) wat hul groei bevorder en hul in staat stel om alkoholiese fermentasie suksesvol te voltooi. Baie van hierdie voedingstowwe, veral stikstof, dien ook as voorlopers vir geurkomponente afkomstig van gismetabolisme, soos hoër alkohole, esters en vlugtige vetsure. Die stikstofmetabolisme van gis speel dus ‘n bepalende rol in wynaroma en -kwaliteit. Nie net is die stikstofbron, konsentrasie en tydsberekening van stikstof toevoeging belangrik nie, maar verskeie omgewingsfaktore, asook die genetiese samestelling van die gisras aangewend vir fermentasie, sal bydra tot die fermentasie uitkomste. Die hoofdoel van hierdie werk was om die komplekse interaksies tussen ‘n aantal bydraende faktore te ondersoek; naamlik die stikstofbron, tyd van stikstof toevoeging, gisras en fermentasiematriks. Breedweg het hierdie studie die impak van sewe verskillende stikstofkombinasies, toegedien tot die druiwemos voor of na die aanvang van fermentasie, op die suksesvolle verloop van fermentasie en die produksie van aromakomponente deur nege kommersiële wyngisrasse bepaal. Fermentasies is in sintetiese druiwemos uitgevoer, en ‘n deelversameling van die fermentasies in regte druiwesap te herhaal. Die stikstofbehandelings is ontwerp in ooreenstemming met die algemeen vasgestelde voorkeurvolgorde van S. cerevisiae vir individuele aminosure as stikstofbron onder fermentatiewe kondisies, en die potensiaal van sekere aminosure om mee te doen in metaboliese paaie wat spesifieke aromaverbindings produseer. Die resultate toon dat verskillende stikstofkombinasies tot onverwagte aroma-uitkomste kan lei wat sterk afhanklik is van die genetiese agtergrond van individuele gisrasse en die tyd van stikstof byvoeging. Sekere stikstofbehandelings het konsekwent, vir veelvuldige gisrasse, tot beduidende toenames of afnames in die konsentrasies van spesifieke aromakomponente gelei in vergelyking met die behandeling wat ammonium as enigste stikstofbron bevat het. Hierdie verbindings is as stikstofbehandeling afhanklik geklassifiseer. Ander aromaverbindings is soortgelyk vir alle stikstofbehandelings geproduseer en is aangewys as stikstofbehandeling onafhanklik. Die teenwoordigheid van spesifieke aminosuurgroepe (byvoorbeeld die vertakte-ketting en aromatiese aminosure) kon gekorreleer word met beduidende veranderings in produksiepatrone van verwante (soos hoër alkohole) of onverwante (dietielsuksinaat) aromakomponente relatief tot die ander stikstofbehandelings. Alles inaggenome kon ‘n aantal interessante en nuwe hipoteses rakende die betrokke metabolise padweë van die data afgelei word. Uiteindelik sal hierdie aanvaklike bepaling van interaktiewe effekte tydens fermentasie bydra tot praktiese riglyne vir wynmakers, wat hulle in staat sal stel om druiwesapbestanddele (soos nutriënte) te strook met die intrinsieke aromaproduksie kapasiteite van spesifieke gisrasse, en sodoende wynaroma, styl en kwaliteit te moduleer. / Oenobrands and THRIP for funding this project

Thesis (MScAgric)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: Wine cellars are diverse in terms of equipment types and process configurations. Whilst other food production processes have, in many cases, been properly analysed and modelled, this process diversity has resulted in an absence of process analyses in the wine industry. Each wine cellar is unique and represents a fully integrated agro-business, starting with a raw material (grapes) and extending to marketing and selling of the final product (wine). This makes the wine industry unique in this context. This study is the first attempt to analyse winemaking procedures in the form of a process audit. The study was approached in the following manner: • A questionnaire was developed to assess cellar configurations and conditions. This questionnaire was submitted to a statistically significant number of cellars, and a statistically significant number of questionnaires were returned. • The data collected from the questionnaire were statistically analysed and associations between equipment or procedures and wine faults were identified. • Three cellars were studied in depth. These three cellars had their processes audited and their effluent characterised. Additional data were obtained from current sampling projects and these data were analysed to complement the data obtained from the questionnaire • A preliminary input/output model was developed. The major results of this study are: • It was found that certain faults that appear in wine might be associated with equipment and/or process faults. These associations are statistically significant and they show that cellar hygiene is of critical importance when assessing these wine faults. The most important of these faults are VA, microbial contamination of the wine, sluggish and stuck fermentations. A risk hierarchy was derived to indicate which events are associated with others most strongly. • It was found that few wineries measure water consumption and even fewer wineries measure the quantity of effluent produced. • Correlations have been developed to predict winery parameters in terms of tons of grapes pressed per annum. These parameters include water and electricity consumed, wine produced and the quantity of effluent produced. Effluent characteristics have also been correlated to the tons of grapes pressed per annum. These characteristics include chemical oxygen demand, sodium absorption ratio and total dissolved solids in solution. Chemical oxygen demand was identified as the most important contributing factor in winery effluent. It was shown that all variables rise with an increase in cellar size, but the rise is not linear. This implies that large cellars have greater quantities of effluent of lower quality than small cellars. Most cellars have effluent concentrations that require some form of effluent treatment. The characterisation of effluent shows that the most widely used disposal practice is irrigation, and that the effluent disposed in this manner does not meet legislative requirements. • A preliminary input/output model was developed in order to enable wineries that have not measured the relevant parameters to predict the abovementioned variations. The resolution of these predictions is low but the model serves to provide an initial estimate if there are no data available. The model will give industrial averages for any given cellar size. • An economic balance was performed using this preliminary model. It was shown that if cellars were to lower the consumption of utilities and to reduce the strength of their effluent (using cleaner practices and not dilution) the reduction of operating costs could be reduced by 14% for smaller cellars to 17% for larger cellars. This study has shown that it is possible to make wine in a more environmentally friendly manner, producing better quality wines, without incurring extra costs. / AFRIKAANSE OPSOMMING: Wynkelders is diverse eenhede ten opsigte van die tipe toerusting en prosesse wat gebruik word. Terwyl ander prosesse rondom voedselproduksie in baie gevalle reeds geanaliseer en gemodelleer is, het die diversiteit in die wynindustrie gelei tot 'n afwesigheid van prosesanalises. Elke wynkelder is uniek en verteenwoordig 'n ten voile geintegreerde agro-besigheid wat begin met die rou materiaal (druiwe) en lei tot die bemarking en verkope van die finale produk (wyn), 'n eienskap wat die wynindustrie uniek maak in hierdie verband. Hierdie studie is die eerste poging om die wynmaakproses in die vorm van ‘n proses-oudit te analiseer. Die studie is soos volg aangepak: • ‘n Vraelys is ontwikkel om kelderkonfigurasies en -toestande te ondersoek. Die vraelys is aan ‘n statisties betekenisvolle aantal kelders voorgele, en ‘n statisties betekenisvolle aantal vraelyste is terugontvang. • Die data wat uit die vraelyste ontvang is, is statisties ontleed en verwantskappe tussen toerusting of prosesse en wyngebreke is geidentifiseer. • Drie kelders is in diepte bestudeer. Hierdie drie kelders se prosesse is geoudit en die afvloeiwater is gekarakteriseer. Addisionele data is verkry van huidige projekte en hierdie data is ge-analiseer om die data van die vraelys aan te vul. • ‘n Voorlopige inset / uitset model is ontwikkel. Die belangrikste resultate van hierdie studie is: • Dit is bevind dat sekere gebreke wat in wyn voorkom geassosieer kan word met tekortkominge in toerusting en/of prosesse in die kelder. Hierdie assosiasies is statisties betekenisvol en toon dat kelderhigtene van kritiese belang is wanneer gebreke in wyn ondersoek word. Die mees belangrike gebreke wat voorkom is vlugtige suur, mikrobiese kontaminasie van wyn, slepende en gestaakte fermentasie. ‘n Risikohierargie is afgelei om die gebeure te toon wat die sterktste met mekaar geassosieer word. • Dit is gevind dat min wynkelders waterverbruik meet. Selfs minder kelders meet die hoeveelheid afvloeiwater wat geproduseer word. • Korrelasies is ontwikkel om kelderparameters te voorspel in terme van ton druiwe gepars per jaar. Hierdie parameters sluit in water- en elektrisiteitsverbruik, wyn geprosuseer en hoeveelheid afvloeiwater geproduseer. Eienskappe van afvloeiwater is ook gekorreleer met die ton druiwe wat per jaar gepars word. Hierdie eienskappe sluit in chemiese suurstofbehoeftes, natrium absorpsieverhoudings en totale opgeloste soliede materiaal in oplossing. Chemiese suurstofbehoeftes is geidentifiseer as die mees belangrike bydraende faktor tot afvloeiwater in kelders. Dit is getoon dat alle veranderlikes verhoog hoe groter die kelder, maar hierdie verhoging is nie lineer nie. Dit impliseer dat groter kelder meer afvloeiwater van ‘n laer kwaliteit produseer. Die meeste kelders produseer afvloeiwater van sodanige konsentrasies dat behandeling daarvan nodig is. Die ondersoek van hierdie afvloeiwater toon dat die mees algemene wyse van wegdoening van afvloeiwater besproeiing is, en dat afvloeiwater wat op hierdie manier weggedoen word, nie voldoen aan die wetgewing se vereistes nie. • 'n Voorlopige inset/uitset model is ontwikkel om kelders wat nie die toepaslike parameters gemeet het nie in staat te stel om hierdie parameters te voorspel. Die akuraatheid van hierdie voorspellings is nie hoog nie, maar die model verskaf ‘n aanvanklike skatting waar daar geen data beskikbaar is nie. Die model verskaf industriele gemiddeldes aan kelders van enige grootte. • ‘n Ekonomiese balans is uitgevoer deur van hierdie model gebruik te maak. Dit is getoon dat indien kelders die gebruik van water en elektrisiteit verminder en die konsentrasie van afvloeiwater verlaag (deur van skoner paktyke gebruik te maak, en nie verdunning nie) die bestuurskoste met 14% vir kleiner kelders tot 17% vir groter kelders verlaag kan word. Die studie het getoon dat dit moontlik is om wyn te maak op ‘n meer omgewingsvriendelike wyse, en sodoende beter kwaliteit wyn te produseer sonder addisionele kostes.

Thesis (MScAgric)--Stellenbosch University, 1952. / No Abstract Available

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: There is a growing international consumer demand for the production of lower ethanol wines. This can be attributed to various qualitative, social, economic and health concerns that are associated with high ethanol wines (Kutyna et al., 2010; Varela et al., 2012). There is continuous development and research into methods and technologies to lower the ethanol concentration in wine. However, in addition to the added cost and complexity these technologies all have various shortcomings. The development of yeast strains with lower ethanol productivity, yet desirable organoleptic and fermentation capacity, therefore remains a highly sought after research and development target in the wine industry. Biologically based approaches aim to generate yeast strains with the capacity to divert carbon from ethanol production towards targeted metabolic endpoints (Kutyna et al., 2010). This should ultimately be achieved without the production of unwanted metabolites that can negatively affect wine characteristics. In the context of these challenges, this study aimed to investigate the use of fructans as carbon sinks during fermentation to divert fructose from glycolysis and ethanol production toward intracellular fructan production by generating levan producing strains. In addition, the impact of fructan production on metabolic carbon flux during fermentation by these strains was analyzed. This was the first attempt to analyze intracellular fructan production in Saccharomyces cerevisiae under fermentative conditions with fructans acting as carbon sinks. Fructans are fructose polymers that act as storage molecules in certain plants and function as part of the extracellular matrix in microbial biofilms, and are intensively studied due to their economic interest. Here we undertook the heterologous expression of a levansucrase (LS) M1FT from Leuconostoc mesenteroides, an enzyme producing β(2-6) levan-type fructans, in the S. cerevisiae BY4742Δsuc2 strains without invertase activity (encoded by SUC2). Levansucrases indeed utilize sucrose as both fructose donor and initial polymerization substrate, and the sucrose concentration is of import to maintain transfructosylation activity of enzyme. High intracellular sucrose accumulation was achieved by the heterologous expression of either a sucrose synthase (Susy; cloned from potato) or by growing strains expressing the spinach sucrose transporter (SUT) in sucrose containing media. Endogenous sucrose synthesis was of specific interest to the overall goal of the project, which was to reroute carbon flux away from glycolysis in grape must containing only hexoses as carbon source. In addition, this approach of combining intracellular sucrose production with intracellular levan production could be used in various applications to limit the need for sucrose in media as both carbon source and LS substrate. The extracellular LS M1FT was introduced into Susy and SUT strains as either the complete gene (M1FT) or 50bp truncation (M1FTΔsp) without the predicted signal peptide. The data show that intracellular levan accumulation occurred in aerobic, but not anaerobic conditions. The data also suggest that the production of levan did not impact negatively on general yeast physiology or metabolism in these conditions. However, no significant reduction in ethanol yields were observed, suggesting that further optimisation of the expression system is required. This is the first report of levan synthesis by S. cerevisiae, and contributes towards expanding the possibilities for further industrial applications of these compounds. / AFRIKAANSE OPSOMMING: Daar is toenemende aanvraag deur wynverbruikers na laër alkohol wyne. Hierdie neiging kan toegeskryf word aan verskeie kwalitatiewe, gesondheids en sosio-ekonomiese redes wat geassosieer word met die verbruik van hoër alkohol wyne. Daar is ’n deurlopende navorsing dryf toegespits op metodes en tegnologieë om die alkohol konsentrasie van wyne te verlaag. Hierdie tegnologieë het egter, bykomstig tot koste en kompliksiteits toename, verkeie tekortkominge. Die ontwikkeling van gisrasse met verlaagde alcohol produksie, maar steeds wenslike organoleptiese en fermentasie eienskappe, bly ‘n baie gesogte navorsings en ontwikkeling teiken in die internasionale wyn industrie. Biologiese benaderings streef om gisrasse te genereer met die vermoë om koolstof weg van etanol produksie te herlei na geteikende metabolise eindpunte. Hierdie doelwit moet ook uiteindelik bereik word sonder die produksie van ongewenste metaboliete wat die wyn negatief kan affekteer. In die konteks van hierdie uitdaging, het hierdie studie gestreef om die gebruik van fruktane as ’n koolstof poel tydens fermentasie, met die doel om fruktose te herlei vanaf glikolise en etanol produksie na intrasellulêre fruktane produksie. Om hierdie doelwit te bereik, is gisrasse ontwikkel wat levaan (’n spesifieke fruktaan) produseer. Die impak van fruktaan produksie op metaboliese koolstof vloei tydens fermentasie deur hierdie gisrasse is bykomsrig ontleed. Hierdie verslag beskryf die eerste poging om intraselullêre fruktaan produksie in Saccharomyces cerevisiae te bewerkstellig, met die doel om fruktaan as ’n koolstof poel te gebruik. Fruktane is fruktose polimere wat as bergings molekules optree in sekere plante en ook funksioneer as deel van die ekstrasellulêre matriks in mikrobiese biofilms. Hierdie polimere word tans internasionaal intensief bestudeer weens hul ekonomiese belang. Hierdie studie beskryf die uitdrukking van die levaansukrase (LS) M1FT van Leuconostoc mesenteroides, wat β(2-6) levaan-tipe fruktane produseer, in S. cerevisiae BY4742Δsuc2 rasse, sonder invertase (gekodeer deur SUC2). Levaansukrases gebruik inderdaad sukrose as beide ’n fruktose donor en ook as ’n aanvanklike polimeriserings substraat. Die fruktose konsentrasie is belangrik om transfruktosilerings aktiwiteit van die ensiem te handhaaf. Hoë intrasellulêre sukrose akkumulasie was bereik deur die heteroloë uitdrukking van ’n sukrose sintase (Susy; gekloneer van aartappel) of die spinasie sukrose transporter (SUT) in media bevattende sukrose. Endogene sukrose sintese was van spesifieke belang tot die algehele doelwit om koolstof te herlei, weg van glikolise tydens fermentase van druiwe sap. Die benadering om intraselullêre sukrose produksie met levaan produksie te koppel, kan ook gebruik word in verskeie toepassings om die afhanklikheid op sukrose in die media, as substraat vir LS, te verminder. Die ekstraselullêre LS, M1FT, was as vollengte geen (M1FT) of as ’n 50bp afkapping (M1FTΔsp), sonder seinpeptied, in die Susy en SUT gisrasse uitgedruk. Die data dui aan dat die produksie van levaan nie ’n negatiewe impak het op gis fisiologie of metabolisme in die toets kondisies nie. Daar was egter geen waarbeenbare afname in etanol opbrengs nie, wat aandui dat verdere optimisering van ekspressie sisiteem benodig word. Hierdie is die eerste verslag van levaan sintese in S. cerevisiae en dra by tot die uitbreiding van moontlikhede vir indutriële toepassings van die die verbindings. / IWBT and NRF

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Proteins from various sources, including grape berry cells, yeast, bacteria and fining agents e.g. albumin and casein, have previously been identified in wine. These proteins play various critical roles in the functioning and survival of the organisms that produced them but also exhibit oenological properties, once secreted in the juice/wine. Some of them can indeed be beneficial to winemaking, by releasing aroma compounds from grape-derived precursors, or detrimental to wine quality, by causing protein haze. Yeasts contribute significantly to the protein pool during and after alcoholic fermentation. However, while the extracellular proteins of Saccharomyces cerevisiae, the main wine yeast species, have been characterised, those of non-Saccharomyces yeasts remain largely unknown, especially under winemaking conditions. Although specific extracellular enzymes released by non-Saccharomyces yeasts have been the focus of many studies in recent years, the targeted approaches used have restricted our knowledge to these specific enzymes and excluded the other secreted proteins. A more comprehensive insight into entire secretomes could improve our understanding of how yeasts survive in wine and interact with other species in mixed culture fermentations. This study aims to characterise the exo-proteome of Saccharomyces and selected non-Saccharomyces yeasts in pure and mixed cultures in a wine-like medium. Fermentation kinetics were monitored and the extracellular proteins isolated at the end of fermentation. M. pulcherrima hardly fermented whereas L. thermotolerans fermented slowly but steadily. As expected S. cerevisiae completed the fermentation rapidly. In sequential fermentations, the kinetics resembled those of the non-Saccharomyces yeasts for a period before switching to that of S. cerevisiae. This period varied from 4 to 15 days for M. pulcherrima and L. thermotolerans respectively. Visual observations of the protein content of the medium at the end of fermentation using 1D and 2D SDS-PAGE gels as well as identification of these proteins using mass fingerprinting revealed the large variety of proteins secreted and the influence of yeast interactions on each other’s secretome. The fermentation kinetics observed could partially be explained by the extent of the contribution of the different yeast to the protein content. Proteins secreted by non-Saccharomyces yeasts lowered the potential of wine to form protein haze, with both M. pulcherrima and L. thermotolerans in pure and mixed culture fermentations showing lower haze formation than S. cerevisiae. As far as we know, this is the first report on the secretome of non-Saccharomyces under winemaking condition and the influence non-Saccharomyces proteins have on the protein haze potential of wine, providing the basis for future investigations. / AFRIKAANSE OPSOMMING: Proteïene vanaf verskeie bronne (insluitend druiwe korrels, gis, bakterieë en verhelderings agente bv. albumien en kaseïen) is reeds in wyn identifiseer. Hierdie proteïene speel verskeie rolle in die funksionering en oorlewing van die organismes wat dit produseer, maar beskik ook oor wynkundige eienskappe sodra dit in die sap of wyn uitgeskei word. Hoewel sommige proteïene in wyn wel voordelig mag wees as gevolg van die vrystelling van aroma komponente vanuit druif‐voorlopers, kan dit ook nadelig wees vir wyn kwaliteit deur die troebelheid wat dit kan veroorsaak Gis dra aansienlik by tot die totale proteïen inhoud van wyn, beide gedurende asook na alkoholiese fermentasie. Alhoewel die ekstrasellulêre proteïene van Saccharomyces cerevisiae (die mees algemeen gebruikte gis vir wynmaak) reeds goed gekarakteriseer is, is die proteïene van nie-Saccharomyces giste grootliks onbekend, veral die wat tydens wynmaak vrygestel word. Gedurende die laaste paar jaar het verskeie studies gefokus op spesifieke ekstrasellulêre ensieme wat deur nie-Saccharomyces giste produseer word, maar geteikende benaderings het ons kennis beperk tot net hierdie spesifieke ensieme, en enige ander afgeskeide proteïene uitgesluit. ʼn Meer omvattende insig oor die algehele afgeskeide proteoom kan ons begrip van hoe gis in wyn oorleef en interaksies tussen gis spesies in gemengde kultuur fermentasies verbeter Hierdie studie streef om die sekretoom van Saccharomyces en geselekteerde nie-Saccharomyces giste in suiwer en gemengde kultuur fermentasies van sintetiese wyn medium te karakteriseer. Fermentasie kinetika is gemonitor en die ekstrasellulêre proteïene is teen die einde van fermentasie geïsoleer. Metschnikowia pulcherrima het swak fermenteer terwyl Lachancea thermotolerans stadig tog reëlmatig fermenteer het. Soos verwag, het S. cerevisiae vinnig tot droog fermenteer. In agtereenvolgend geïnokuleerde fermentasies is die kinetika vir ʼn tydperk soortgelyk aan die van die nie-Saccharomyces giste voordat dit oorskakel na die van S. cerevisiae. Hierdie tydperk wissel respektiewelik vanaf 4 tot 15 dae vir M. pulcherrima en L. thermotolerans. Visuele waarnemings van die proteïen-inhoud van die medium aan die einde van die gisting met behulp van 1D en 2D SDS-PAGE gels asook identifisering van hierdie proteïene met behulp van massa vingerafdrukke onthul die groot verskeidenheid proteïene wat afgeskei word, asook die invloed van die giste se interaksies op mekaar se sekretoom. Die fermentasie kinetika waargeneem kan gedeeltelik verklaar word deur die omvang van die bydrae van die verskillende gis tot die proteïen-inhoud. Proteïene wat afgeskei word deur nie-Saccharomyces giste verlaag die potensiaal van wyn om proteïen troebelheid te vorm, met beide M. pulcherrima en L. thermotolerans (in suiwer en gemengde kultuur fermentasies) wat minder troebelheid vorm as fermentasies met S. cerevisiae. Sover ons kennis strek, is hierdie die eerste verslag oor die sekretoom van nie- Saccharomyces onder wynmaak toestande en ook oor die invloed wat nie-Saccharomyces proteïene op die proteïen troebelheid van wyn het, en vorm die basis vir toekomstige navorsing. / Winetech and THRIP

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Nitrogen composition of grape must is highly variable and impacts on the health of the fermenting yeast population as well as the formation of aroma and flavour compounds in wine. Insufficient yeast assimilable nitrogen (YAN), mostly consisting of amino acids and ammonium, can lead to stuck or sluggish fermentations as well as the formation of undesirable compounds such as H2S. Furthermore, it is well established that the total concentration of YAN and the specific amino acid composition have a significant impact on the final aroma and flavour of wines. However, the impact of individual amino acids and of specific amino acid compositions on fermentation kinetics and on the production of aroma and flavour impact compounds under winemaking conditions is not well understood. The first goal of this study was to evaluate the effect of single amino acids on growth kinetics and major volatile production of two industrial wine yeast strains under conditions resembling wine fermentations. To facilitate these fermentation conditions while also allowing for easy reproducibility and manipulation of the initial components, a synthetic grape media was utilized. Biomass formation, exponential growth rate, lag phase, and fermentation duration were utilized to evaluate the efficiency of single amino acids. The data show that previously observed trends in laboratory strains mostly apply to these conditions and strains. In general, the efficiency of amino acids to be used as nitrogen sources and the production of major volatiles due to their presence followed the same patterns for both industrial yeast strains. However, the production of the secondary metabolites butanol, propanol, acetic acid, and ethyl acetate were found to be produced in different final concentrations dependent upon the yeast strain. The branched-chained and aromatic amino acids (BCAAs) treatments were observed to have the most dramatic effects on major volatile production. Investigating the relationships between the initial concentration of the BCAAs and the final concentrations of major volatile compounds, it was found that the production of fusel alcohols and fusel acids due to the degradation of BCAAs by S. cerevisiae could be predicted from the initial concentration of BCAAs. While under simple nitrogen conditions the production of several other secondary metabolites such as butanol, propionic acid, valeric acid, decanoic acid and 2-phenylethyl acetate were found to be correlated to the initial concentration of BCAAs in the media. Future studies should focus on the validation of these trends in aroma production in real grape musts under various fermentation temperatures for a number of industrial wine yeast strains. / AFRIKAANSE OPSOMMING: Die stikstof samestelling van druiwemos is hoogs veranderlik en impakteer op die kerngesondheid van die fermenterende gis populasie asook die produksie van aroma- en geurverbindings in wyn. Onvoldoende assimileerbare stikstof (ook genoem “yeast assimilable nitrogen” (YAN)), wat meestal bestaan uit aminosure en ammonium, kan aanleiding gee tot steek- of slepende fermentasies, asook die vorming van ongewensde verbindings soos H2S. Dit is alombekend dat die totale konsentrasie van YAN en dan ook die spesifieke aminosuur samestelling ‘n noemenswaardige impak op die finale aroma en smaak van wyn het. Die invloed van individuele am inosure en spesifieke aminosuur samestellings op die fermentasie kinetika, asook die produksie van verbindings met ‘n impak op die wyn aroma en smaak, word egter nie deeglik verstaan nie. Die eerste doelwit van die studie was om twee industriële gisrasse te gebruik om die effek van enkel aminosure op die groei-kinetika en produksie van die vername vlugtige verbindings onder wynmaak toestande te bepaal. Kunsmatige, gedefinieerde druiwermosmedium is gebruik om wynmaak toestande te simuleer en ook herhaalbaarheid en manipulering van die aanvanklike samestelling van die medium te verseker. Die studie het vorige tendense wat opgemerk is in die evaluasie van laboratorium rasse onder soortgelyke toestande bevestig. Die doeltreffendheid waarmee aminosure oor die algemeen gebruik word as stikstofbron, asook die produksie van die vernaamste vlugtige verbindings wat gekoppel is aan hulle teenwoordigheid, het ‘n vergelykbare patroon vir beide rasse gevolg. Die sekondêre metaboliete butanol, propanol, asetaat en etiel-asetaat is egter wel in verskillende eindkonsentrasies geproduseer deur die verskillende gisrasse. Die vertakte-ketting en aromatiese aminosuur (“branched-chained and aromatic amino acids” (BCAAs)) behandelings het die mees dramatiese effek op die produksie van die vernaamste vlugtige komponente gehad. Ondersoek na die verwantskap tussen die aanvanklike konsentraies van die BCAAs en die finale konsentrasies van dié verbindings het aangedui dat die produksie van hoër alkohole en sure, as gevolg van die afbraak van BCAAs deur S. cerevisiae, met behulp van die aanvanklike konsentrasie van die BCAAs voorspel kon word. Terselfdertyd is gevind dat onder eenvoudige stikstoftoestande, verskeie ander sekondêre metaboliete soos butanol, propionaat, valeriaat, dekanoësuur en 2-fenieletielasetaat, gekorreleer kan word met die aanvanklike BCAAs in die media. Verdere studies moet poog om hierdie tendense ten opsigte van aromaproduksie te bevestig en wel deur gebruik te maak van ware druiwemos, verskeie fermentasietemperature, asook ’n verskeidenheid van wyngisrasse.

Thesis (MScAgric)--University of Stellenbosch, 2006. / Bibliography. Also available via the Internet.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The alcoholic beverage industry needs objective analysis of grape juice and liqueur quality. Fourier transform infrared (FT-IR) spectroscopy with multivariate data analysis techniques is widely used in wine laboratories across South Africa for accurate, fast and high sample throughput analyses. In this study the potential of FT-IR spectroscopy is evaluated for the quantification of ammonia in freshly pressed grape juice. FT-IR spectroscopy is evaluated, using two different spectrometers, in respectively attenuated total reflection (ATR) - and transmission scanning modes for the quantification of alcohol, pH and invert sugar in spirit-based liqueurs. The ultimate aim was to implement the PLS regression algorithms developed at an industrial cellar and replace the complex and lengthy reference methods used at the time of this study. Principle component analysis (PCA) was performed prior to the calibration step to identify groupings and patterns within the spectra. The PLS calibration models were developed from samples collected at the cellar and using partial least square (PLS) regression. The models were evaluated using the performance criteria coefficient of determination (R2) and root mean squared error of cross validation (RMSECV) at calibration stage, and root mean square error of prediction (RMSEP) and residual predictive deviation ratio (RPD) at validation stage. The average RMSEP (1.88 mg/L) of the ammonia PLS calibration model was in agreement with the standard error of laboratory (SEL = 1.54 mg/L). The R2 (92.05) and average RPD (3.3) proposed a model with excellent precision for screening purposes that was ready to be transferred for use by the laboratory. The r2 values for the alcohol, pH and invert sugar PLS calibration models obtained in ATR and transmission, indicated good to excellent precision (80<r2<100). The alcohol PLS calibration model obtained in transmission was suitable for quality- and process control purposes (RPD = 21.2), while the invert sugar PLS calibration model for quality control purposes (5<RPD<6.4). The pH and invert sugar calibration models obtained in ATR were suitable for screening purposes with RPD = 3.6 and RPD = 4.8, respectively. These PLS regression algorithms were implemented at the cellar. The pH PLS calibration model obtained in transmission was suitable for rough screening of samples (RPD = 2.7) and future development was neccesary to increase the predictability of the model. The results obtained in this study made a significant contribution towards validation of FT-MIR as a powerful tool for rapid quantification of quality indicating parameters in wine and spirit-based liqueurs. The contribution is particularly valuable in the context of ongoing research to improve the quality of products at the cellar to meet consumer demands. The knowledge gained on quantification of quality indication parameters of spirit-based liqueurs is novel and this is one of the first reports on implementation of mid-infrared (MIR) spectroscopy for the quality control of South African spirit-based liqueurs. / AFRIKAANSE OPSOMMING: Die wynindustrie benodig objektiewe analises van druiwesap- en likeurgehalte. Fourier-transformasie- infrarooi (FT-IR) spektroskopie met multiveranderlike statistiese metodes word gebruik in wynlaboratoriums regoor Suid-Afrika vir akkurate, vinnige en hoë monsterdeurset ontledings. In hierdie studie is die potensiaal van FT-IR spektroskopie geëvalueer vir die kwantifisering van ammoniak in die sap van vars geparste wyndruiwe. Twee verskillende FT-IR spektroskopie instrumente, in onderskeidelik (verswakte totale refleksie, ATR) - en transmissie skandering is gebruik vir die kwantifisering van alkohol, pH en invertsuiker in spiritus-gebaseerde likeurs. Die uiteindelike doel was om die parsiële kleinste kwadraat (PKK)- regressie algoritmes wat ontwikkel is, by 'n industriële kelder te implementeer en die komplekse en tydrowende verwysingmetodes wat tydens die studie in die kelder gebruik is te vervang. Verskeie multiveranderlike hoofkomponentanalise (MVK) is uitgevoer voor die kalibrasie stap, met die doel om groeperings en patrone in die spektra te identifiseer. Die PKK kalibrasiemodelle is ontwikkel van monsters wat by die kelder versamel is en die spektra is gebruik in die PKK regressies. Tydens die kalibrasiefase is die modelle geëvalueer met behulp van die bepalingskoëffisiënt (R2) en gemiddelde kalibrasieprediksiefout en tydens die validasiefase, met behulp van die standaardvoorspellingsfout (SVF) en relatiewe voorspellingsafwyking (RVA). Die gemiddelde SVF (1.88 mg/L) van die ammoniak kalibrasiemodel was in ooreenstemming met die standaard fout van die laboratorium (SEL = 1.54 mg/L). Die R2 (92.05) en die gemiddelde RVA (3.3) dui op ‘n model met uitstekende presiesheid wat gereed is vir oordra en gebruik deur die industrie. Die R2 waardes vir die alkohol-, pH- en invertsuiker –kalibrasie-modelle wat met ATR en transmissie vir die likeurmonsters ontwikkel is, dui op goeie tot uitstekende presiesheid (80<R2<100). Die alkoholkalibrasiemodel wat ontwikkel is in transmissie, is geskik vir kwaliteits- en prosesbeheerdoelwitte (RVA = 21.2), terwyl die invertsuiker kalibrasiemodel geskik is vir kwaliteitsbeheer doelwitte (5<RVA<6.4). Die pH en invertsuiker kalibrasiemodelle in ATR is geskik vir vinnige evalueringsdoelwitte, met RVA = 3.6 en RVA = 4.8 waardes, onderskeidelik. Hierdie algoritmes is ook in die kelder geimplementeer. Die pH kalibrasiemodel in transmissie was geskik vir vinnige evalueringsdoelwitte (RVA = 2.7) en toekomstige ontwikkeling is nodig om die voorspellingsakkuraatheidvan die model te verbeter. Die resultate van hierdie studie het ‘n betekenisvolle bydrae gelewer tot bevestiging van infrarooi spektroskopie as 'n kragtige tegnologie vir die vinnige kwantifisering van gehalteparameters in druiwesap en spiritus-gebaseerde likeurs. Die bydrae is veral waardevol in die konteks van voortgesette navorsing om die kwaliteit van produkte by die kelder te verbeter en aan verbruikerseise te voldoen. Die studie vir die kwantifisering van gehalteparameters in spiritus-gebaseerde likeurs is eerste in sy soort en een van die gerapporteerde verslae vir die implementering van infrarooi spektroskopie vir gehaltebeheer van Suid-Afrikaanse spiritus-gebaseerde likeurs.

Thesis (MScAgric)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The quality of wine is largely dependent on environmental conditions and recent studies have therefore focused on agricultural practices in terms of water, soil and biodiversity conservation. The industry aims to create sustainable practice and to protect the natural resources available. Sources of potential grape contamination include: vehicle pollution, pesticides, bushfires and wood preservatives used for trellising systems. The latter have come to the attention of the South African wine industry (e.g. creosote and Copper Chromium Arsenate (CCA) products) as they may have consequences for the environment and wine quality. Creosote is a known pollutant of soil and ground water and the volatile fraction has been monitored in air. Plants may also accumulate polycyclic aromatic hydrocarbons (PAHs), which constitute up to 85% of the mass of creosote, and of which some have been proven carcinogenic. Because of the health risks associated with it, creosote has therefore been restricted for use in most applications in Europe, and also in the United States, Canada and Australia. This study focussed on the sensorial and chemical analyses of environmental and wine samples taken from the area around the creosote stockyard to determine accumulation of creosote-derived compounds. Environmental samples were collected and analysed at different distances from the affected area, over two vintages. Wines were made from grapes grown in vineyard blocks adjacent to the stockyard, to determine the effect of distance and skin contact during alcoholic fermentation treatments on wine taint. A sensory panel was trained for descriptive analysis to determine the intensity of the taint. Analytical methods were developed for the analysis of volatile organic compounds (VOCs) by gas chromatography mass spectrometry (GC-MS) and polycyclic aromatic hydrocarbons by high performance liquid chromatography with diode array detection (HPLC-DAD). The sensory results obtained showed white and rosé wines were perceived as clean, whilst the red wines were associated with burnt rubber/tar taint. The perceived taint decreased as distance from the stockyard increased. Wines made from the Cabernet Sauvignon blocks adjacent to the stockyard also showed an increase of taint with the increase of skin contact. Chemical data obtained showed that the taint consisted of a complex mixture of compounds, each with its own pattern of retention within the vineyard and wine. Only m-cresol was found above odour threshold, and only in red wines. The synergistic effect of some compounds may lead to an increase in the perceived taint. Berries and leaves had higher concentrations of volatile compounds than wines. Leaf contamination varied and followed the general trend in literature where the plants with high lipid content and exposed leaf area were the most contaminated. There may be other compounds present in creosote emissions, aside from those investigated here, with sensory attributes related to the taint found in wines. These compounds are styrene, indene, benzene, toluene, isoquinoline and quinoline and should be included in further investigations. From the PAH analysis of environmental samples and wines, it is evident that the samples closest to the stockyard were affected the most. The contamination varied with the depth of the soil: some of the heavier compounds were found in the samples taken from the deeper levels, whilst nearly all other target compounds were present in the top layer of soil. The concentrations found in the environmental samples were lower than reported in literature. Wines had few PAHs present, but at much higher concentrations than is allowed by EU legislation. From this study it is evident that the stockyard had negative effects on the surrounding environment in terms of sensory and chemical contamination. Recommendations include area rehabilitation by means of bioremediation to protect resources and ensure sustainable and safe production of crops. Industrial emissions should also be regulated and restricted in agricultural areas. Furthermore the use of creosote should be revised, and prohibited for agricultural use. / AFRIKAANSE OPSOMMING: Die kwaliteit van wyn is grootliks afhanklik van die omgewingstoestande. Daarom fokus verskeie onlangse studies op landbou aktiwiteite en die invloed daarvan op die omgewing in terme van water, grond en biodiversiteit bewaring. The wyn industrie se doelwitte sluit volhoubare praktyke in, wat die natuurlike hulpbronne bewaar en beskerm. Druiwe kan deur middel van die volgende bronne besoedel word: brandstof uitlaatgasse, insekdoders, veldbrande, gifstowwe wat gebruik word vir houtperservering wat dan verder aangewend word vir opleistelsels. Houtperservering (Koper chroom arsenaat (CCA) en kreosoot) het veral in die laaste paar jaar onder aandag gekom in die wynbedryf van Suid-Afrika as gevolg van die invloed daarvan op die omgewing en die uiteindelike wynkwaliteit. Kreosoot is ‘n gekende gifstof wat verantwoordelik is vir grond en grondwater besoedeling en wat gemonitor word in die atmosfeer. Plante akkummuleer ook poli-sikliese aromatiese hidro-koolstowwe (PAHs), wat to 85% uitmaak van die massa van kreosoot. PAHs is karsinogenies en dus is daar baie navorsing op die molekules reeds gedoen. Die gesondheidsrisikos gepaardgaande met kreosoot het gelei tot die streng geregulasies tans ingestel in Europa, die Verenigde State, Kanada en Australië. Hierdie studie het gefokus op die sensoriese en chemiese analises van omgewings- en wyn-monsters geneem van die omliggende area van die kreosoot palewerf om die akkumulasie van kreosoot-afgeleide-verbindings. Omgewingsmonsters was versamel en geanaliseer om verskillende afstande vanaf die bron van besoedeling (palewerf) te ondersoek oor ‘n twee jaar periode. Wyne is gemaak van die druiwe wat afkomstig is van die blok aangeplant langs die palewerf. Die wyne is ondersoek in terme van afstand vanaf die kreosoot bron asook oenologiese invloede, dopkontak gedurende alkoholiese fermentasie, op die kontaminasie beskryf in wyn. Die wyne is ook oor ‘n twee jaar periode voorberei en sluit die 2011 en 2012 seisoen in. ‘n Sensoriese paneel is opgelei om die beskrywende analises op die wyn uit te voer met die doel om die intensiteit van kontaminasie te identifiseer. Analitiese metodes is ook ontwikkel vir die analise van vlugtige organiese verbindings(VOCs) met gas chromatografie-massa spektrometrie (GCMS) en poli-sikliese aromaties hidro-koolstowwe (PAHs) met hoë druk vloeistof chromatografie. Die sensoriese resultate bekom het wit en rosé wyne as skoon laat blyk, terwyl rooi wyne meer geassosieer was met die gebrande rubber/ teer afgeur beskrywing. Die waargeneemde afgeur het afgeneem soos wat die afstand vanaf die palewerf toegeneem het. Wyne gemaak van die Cabernet Sauvignon blokke langsaan die palewerf het ook ‘n toename in die afgeur gehad met ‘n toename in dopkontak. Chemiese data bekom beeld uit dat die afgeur uit ‘n komplekse mengsel van verbindings bestaan, elk met sy eie patroon van verspreiding en verbinding in die wingerd as ook in die wyn. Net m-kresol was gevind bo die reuk drumpel, dit het ook net in rooi wyne voorgekom. Die sinergistiese effek van die verbindings mag egter bydra tot die waargeneemde afgeur. Druiwekorrels en blare het hoër konsentrasies van die vlugtige verbindings gehad as wat gemeet is in die wyne. Blaar kontaminasie het ook baie gewissel en het ooreengestem met die algemene tendens wat in literatuur beskryf is, naamlik dat plante met ‘n hoër lipid inhoud en grootter blaar oppervlak die meeste gekontamineer word. Daar mag egter nog baie ander verbindings bydra tot die waargeneemde afgeur gevind in die wyn. Spesifieke verbindings wat wel ‘n rol kan speel in kontaminasie en wat voorkom in die vlugtige gedeelte van kreosoot is styreen, indeen, benzeen, tolueen, isoquinoleen die vlugtige verbindings van kreosoot. Die verbindings moet ingesluit word vir verdere studies wat gedoen word op die kreosoot geassosieerde afgeur. Die PAHs analise op die omgewingsmonsters en wyne het gelei tot die bevestiging dat die naasliggende omgewing die meeste geaffekteer is. Die kontaminasie wissel in terme van die diepte in die grond wat die gifstowwe voorkom: die swaarder molekulêre verbindings is tot in die dieper vlakke waargeneem terwyl al die gemete verbindings in die boonste lae teenwoordig was. Die vlakke wat waargeneem is in dié studie is egter laer as wat voorheen in literatuur gevind is in ‘n kreosoot geaffekteerde omgewing. Wyn het PAHs teenwoordig gehad, alhoewel slegs twee verbindings gemeet is, het dit in hoër vlakke voorgekom as wat sekere Europese regulasies as toelaatbaar spesifiseer. Vanaf die studie resultate blyk dit, dat die palewerf se negatiewe invloed op die omliggende omgewing beide meetbaar was in sensories en chemiese kontaminasie. Voorstelle sluit onder andere die rehabilitasie van die omliggende omgewing deur middel van bioremediasie in. Om sodoende die natuurlike hulpbronne in die area te bewaar asook om volhoubare en veilige verbouing van gewasse te verseker. Industriële besoedeling en afval moet ook gereguleer word en beperk word in landbou areas. Verder moet die gebruik van kreosoot heroorweeg word en strenger regulasies moet in plek gestel word om aan internastionale standaarde te voldoen. / Financial support from Winetech, Trip and UFS

Thesis (MScAgric)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The influence of malolactic fermentation (MLF) in most red and some white wines is one of many factors that determine or influence wine quality, because it affects the flavour and sensory profile of wine. This process is a decarboxylation process conducted by lactic acid bacteria (LAB) such as Oenococcus, Lactobacillus, Pediococcus and Leuconostoc. Mostly Oenococcus oeni, but recently also Lactobacillus plantarum is used in commercial starter cultures and also the first mixed MLF starter culture (NT 202 Co-Inoculant) was commercialized in 2011. The reason for the predominant use of O. oeni and recently L. plantarum is due to their tolerance to the harsh wine environment. Malolactic fermentation leads to a decrease in acidity and an increase in pH that leaves the wine with a softer mouthfeel. Another reason to conduct MLF is the improvement of microbial stability by the removal of malic acid as carbon source. Research focus has recently shifted to the ability of LAB and MLF as well as the interaction of LAB with yeast to alter the wine aroma profile via the modification and/or production of certain aroma compounds. The main goal of this study was to assess the impact of yeast and nutrient addition on the ability of the NT 202 Co-Inoculant to conduct MLF during co-inoculation and to evaluate the aroma compound production in the final wine. The first aim was to evaluate the impact of different red and white wine yeast strains on the ability of the NT 202 Co-Inoculant to conduct MLF during co-inoculation in Chardonnay, Merlot and Shiraz. Malolactic fermentation was unsuccessful in the Chardonnay due to a low pH, but successful in Merlot and Shiraz. Based on the malic acid degradation ability of the NT 202 Co- Inoculant, the yeasts were grouped into three categories: inhibitory, neutral or stimulatory towards MLF. Co-inoculated MLF showed a clear decrease in total fermentation time while yeast strains such as WE 372 and Exotics showed positive compatibility with the NT 202 Co- Inoculant. The impact of the yeast-bacterial combinations on the aroma compound production in the final wine was evaluated. Co-inoculated MLF showed positive aroma changes in the red wines with a general increase in total esters (associated with fruity characters in wine) especially ethyl lactate and diethyl succinate that also contribute to the mouthfeel of the wine. Production of esters, volatile fatty acids and higher alcohols seemed to depend on the yeastand LAB strain used. The NT 202 Co-Inoculant contributed to the monoterpenes produced and MLF led to increased concentrations of diacetyl and acetoin, which are associated with buttery characters in wine. The second aim of this study was to evaluate the impact of wine additives (used during coinoculation) such as yeast- and bacterial nutrients, clarifying- and detoxifying agents on the ability of the NT 202 Co-Inoculant to conduct MLF and to assess their impact on the aroma compound production in the final wine. No negative or positive impact on the malic acid degradation of the NT 202 Co-Inoculant or the resulting aroma compound production was observed for the different wine additives used in this study. The results generated from this study showed that the selection of yeast strains is important as it will influence both the fermentation duration and final wine aroma. / AFRIKAANSE OPSOMMING: Die invloed van appelmelksuurgisting (AMG) in die meeste rooi- en witwyne is een van baie faktore wat wynkwaliteit beïnvloed, omrede dit die geur en sensoriese profiel van wyn beïnvloed. Hierdie proses is 'n dekarboksileringsaksie wat deur melksuurbakterieë (MSB), soos Oenococcus, Lactobacillus, Pediococcus en Leuconostoc, uitgevoer word. Die mees algemene bakterieë wat gebruik word, is Oenococcus oeni, maar onlangs het Lactobacillus plantarum ook na vore getree in die gebruik van kommersiële aanvangskulture. Die eerste gemengde AMGaanvangskultuur (NT 202 Co-Inoculant) is in 2011 gekommersialiseer. Die rede vir die oorheersende gebruik van O. oeni en L. plantarum word toegeskryf aan hul gehardhiedsgraad in ‘n uitdagende wynomgewing. Appelmelksuurgisting lei tot 'n afname in die suurheidsgraad en 'n toename in die pH van die wyn, wat 'n sagter mondgevoel tot gevolg het. Nog 'n rede waarom AMG deurgevoer word, is om die mikrobiese stabiliteit van die wyn te verbeter deur die verwydering van appelsuur as koolstofbron. Die navorsingsfokus het onlangs verskuif na die vermoë van MSB en AMG, sowel as die interaksie van MSB met die gis, om die wynaromaprofiel te verander deur middel van die verandering en/of produksie van sekere aromaverbindings. Die hoofdoel van hierdie studie was om die impak van die gis en voedingstof te evalueer ten opsigte van die vermoë van die NT 202 Co-Inoculant om AMG uit te voer tydens koïnokulasie. Die produksie van aromakomponente in die finale wyn is ook geëvalueer. Die eerste doelwit was om die impak van verskillende rooi- en witwyngisrasse te evalueer ten opsigte van die vermoë van die NT 202 Co-Inoculant om AMG uit te voer tydens koïnokulasie in Chardonnay, Merlot en Shiraz. Appelmelksuurgisting was onsuksesvol in die Chardonnay weens 'n lae pH, maar suksesvol in Merlot en Shiraz. In terme van die appelsuurafbraakvermoë van die NT 202 Co-Inoculant, is die giste in drie kategorieë gegroepeer: inhiberend, neutraal of stimulerend teenoor AMG. Ge-koïnokuleerde AMG het 'n duidelike afname in die totale fermentasietyd getoon, terwyl gisrasse, soos WE 372 en Exotics, ‘n positiewe verenigbaarheid met die NT 202 Co-Inoculant getoon het. Die impak van die gisbakteriële kombinasies op die aromakomponentproduksie in die finale wyn is geëvalueer. Gekoïnokuleerde AMG het positiewe aromaveranderinge in die rooiwyne getoon met 'n algemene toename in die totale esters (wat geassosieer word met vrugtige karakters in wyn), veral etiellaktaat en dietielsuksinaat, wat ook bydra tot die mondgevoel van die wyn. Dit het voorgekom dat produksie van esters, vlugtige vetsure en hoër alkohole moontlik afhanklik kan wees van die gis- en bakteriële ras gebruik. Die NT 202 Co-Inoculant het bygedra tot die monoterpene wat geproduseer is en AMG het gelei tot verhoogde konsentrasies van diasetiel en asetoïen, wat geassosieer word met botteragtige karakters in wyn. Die tweede doelwit van hierdie studie was om die impak van wyntoevoegingsmiddels (wat tydens koïnokulasie gebruik word) bv. gis- en bakteriese voedingstowwe, verhelderingsagente, asook detoksifiserende agente, te evalueer ten opsigte van die vermoë van die NT 202 Co- Inoculant om AMG uit te voer en om hul impak op die produksie van die aromakomponente van die finale wyn te ontleed. Geen negatiewe of positiewe effekte is waargeneem vir die verskillende wyntoevoegingsmiddels, wat in hierdie studie gebruik is, in terme van die appelsuurafbraak van die NT 202 Co-Inoculant of die gevolglike produksie van aromakomponente nie. Hierdie studie se resultate toon dat die keuse van die gisras belangrik is, omdat dit die fermentasietydperk, asook die finale wynaroma, beïnvloed. / Anchor Yeast, Oenobrands, The National Research Foundation and THRIP, for financial support