Thesis (MSc)--Stellenbosch University, 1965. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MSc)--Stellenbosch University, 1966. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1966. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1960. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1970. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1978. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1950. / No Abstract Available

Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Grapevine performance and wine quality are influenced by various factors, two of the most important being the availability and quality of irrigation water. In relatively dry countries such as South Africa the conservation and effective use of water is of utmost importance. Expected increases in temperature and decreases in rainfall in the future due to climate change impacts highlights the importance of water conservation. This inspired investigations into possible alternative irrigation water sources and therefore the possibility of vineyard irrigation using winery wastewater is of utmost importance for the sustainability of the wine industry. Winery wastewater contains higher concentrations of certain elements other than water generally used for vineyard irrigation, the most important differences being Na and K levels. Furthermore, winery wastewater contains larger populations of microorganisms such as yeasts, lactic acid bacteria and acetic acid bacteria, typical associated with wine production. If irrigation using winery wastewater affects the uptake of certain elements or alters grapevine water status, it may affect grapevine growth, juice and wine composition. Furthermore, if juice and wine composition is affected wine composition and sensorial quality may be affected. Cabernet Sauvignon/99R grapevines, growing in a sandy soil in the Breede River Valley, were subjected to eight irrigation treatments using augmented winery wastewater in addition to irrigation using raw river water as control. The study was carried out during the 2010/11 and 2011/12 seasons. The various wastewater irrigation treatments were made up by augmenting winery wastewater with raw river water to obtain a target chemical oxygen demand (COD) concentration. In this study, the level of COD in the irrigation water is a direct indication of water quality, the two being indirectly proportional. The eight wastewater irrigation treatments ranged from 100 mg/L COD up to 3000 mg/L COD. The first objective of the study was to determine the effect of irrigation using augmented winery wastewater on grapevine response, with regards to vegetative growth, berry development and berry composition. The various wastewater irrigation treatments did not affect grapevine vegetative growth or reproductive growth, including yield, throughout berry development up to harvest. Berry sugar accumulation and evolution in acid concentrations were also not affected. An increase in berry juice pH was observed with an increase in the level of COD in the augmented winery wastewater only in the second season. The amount of elements, ions and heavy metals in juice was not affected by wastewater irrigation, indicating that there was no absorption by the grapevines. Berry skin thickness, colour and phenolic content as well as yield and its associated components were not affected by irrigation using augmented winery wastewater. The second objective of the study was to determine the effect of irrigation using augmented winery wastewater on wine microbial and chemical composition, fermentation performance and wine sensorial characteristics. The natural yeast and bacteria flora of juice was not affected by the various wastewater irrigation treatments. In addition, the ability of the inoculated yeast and lactic acid bacteria strains to conduct their respective fermentation processes were not affected. With the exception of total titratable acidity (TTA) and pH, irrigation using augmented winery wastewater did not affect wine chemical composition with regards to basic wine parameters as well as colour, phenolic and tannin composition. Similar to juice, phosphorus and selected ions were not affected. None of the measured wine sensorial characteristics were affected by irrigation using augmented winery wastewater. The third objective of the study was to investigate the effect of direct contact between berries and winery wastewater on wine sensorial characteristics. The study focussed on the transference of off-flavours from the wastewater into the wine and the occurrence of off-flavours as a response to contact with winery wastewater. Wine colour and general sensory wine descriptives were not affected by direct contact with winery wastewater. The presence of a winery wastewater-like off-odour and volatile acidity was, however, more detectable in wines made from berries that were in contact with the most concentrated wastewater. Therefore, it may be possible for off-odours to be transferred from the winery wastewater into the wines, or that off-odours are formed as a direct or indirect result of contact with winery wastewater. Under the given conditions, results obtained in this two seasons of the study suggest that irrigation using augmented winery wastewater does not affect grapevine performance or wine quality substantially. The major impact that was observed was an increase in wine pH and a decreasing trend in TTA. Both these parameters could be rectified by simply adding acid to the wines. Therefore, irrigation using augmented winery wastewater may be considered as a possible future alternative source for vineyard irrigation. It is, however, important to remember that some of the effects of wastewater irrigation may be cumulative and could possibly arise only after several years. Furthermore, different field conditions and cultivars may respond differently. / AFRIKAANSE OPSOMMING: Wingerd prestasie en wyngehalte word deur verskeie faktore beïnvloed waarvan twee van die belangrikste die beskikbaarheid en gehalte van besproeiingswater is. In relatiewe droë lande soos Suid Afrika is waterbesparing en die effektiewe benutting van water hulpbronne van uiterste belang. Die verwagte toename in temperatuur en afname in reënval in die toekoms as gevolg van klimaatsveranderinge plaasdieklem op op die belangrikheid van waterbesparing. Dit het navorsing om moontlike alternatiewe vorme van besproeiingswater te ontdek geïnspireer. Na aanleiding van hierdie faktore word daar toenemend gefokus op navorsing oor die moontlikheid om kelder afvalwater as alternatiewe bron van besproeiings water vir wingerde te benut. Kelder afvalwater bevat hoër konsentrasie van sekere elemente as water wat onder normale omstandighede gebruik word vir die besproeiing van wingerde, die belangrikste verskille was die vlakke van Na en K. Benewens die hoër konsentrasie van sekere elemente bevat kelder afvalwater ook groot populasies van mikroörganismes soos giste, melksuurbakterieë en asynsuurbakterieë, tipies geassosieerd met wynbereiding. Indien besproeiing met kelder afvalwater die opname van sekere elemente of die plant water status beϊnvloed, mag wingerd groei, sap en wyn samestelling beϊnvloed word. Daar benewens, indien die mikrobiese samestelling van die sap en wyn beϊnvloed word sal die samestelling en sensoriese gehalte van die wyn moontlik beϊnvloed word. Cabernet Sauvignon/99R wingerde, geleë in sanderige grond in die Breede Rivier Vallei, is onderwerp aan besproeiing met agt verskillende konsentrasies van verdunde kelder afvalwater, bykomend tot besproeiing met onbehandelde rivier water wat as kontrole gedien het. Hierdie studie is uitgevoer gedurende die 2010/11 en 2011/12 seisoene. Die teiken besproeiings konsentrasies is verkry deur kelder afvalwater met onbehandelde rivier water te verdun tot ‘n sekere chemiese suurstofbehoefte (CSB) konsentrasie bereik is. Die CSB is in hierdie studie ‘n direkte aanduiding van watergehalte, die twee was indirek eweredig tot mekaar. Die agt CSB konsentrasies waarteen die afvalwater besproei is wissel tussen 100 mg/L CSB en 3000 mg/L CSB. Die eerste doelwit van die studie was om te bepaal wat die effek van besproeiing met verdunde kelder afvalwater op wingerdprestasie, met spesifieke verwysing na vegetatiewe groei, korrelontwikkeling en korrelsamestelling is. Wingerd vegetatiewe en reproduktiewe groei, insluitende opbrengs, is op geen stadium tydens korrelontwikkeling tot en met oes beïnvloed nie. Die laai van suikers gedurende rypwording, sowel as verskuiwings in suurkonsentrasie, is nie deur besproeiing met kelder afvalwater beïnvloed nie. In die tweede seisoen is ‘n toename in sap pH waargeneem soos die CSB konsentrasie van die besproeiings water toegeneem het. Die element, ioon en swaar metaal samestelling van sap was nie beïnvloed deur besproeiing met afvalwater nie wat aandui dat daar geen opname was deur die wingerd nie. Die dikte, kleur en fenoliese samestelling van druifdoppe is ook nie beïnvloed nie. Die tweede doelwit van die studie was om te bepaal wat die effek van besproeiing met verdunde kelder afvalwater op wyn mikrobiese en chemiese samestelling, fermentasie effektiwiteit en wyn sensoriese eienskappe is. Die verskeie afvalwater besproeiings behandelings het geen effek op die natuurlike gis of bakterieë flora van die sap gehad nie. Die vermoë van die geïnokuleerde gis en melksuurbakterieë om hul afsonderlike fermentasie prosesse te voltooi is ook nie beïnvloed nie. Met die uitsondering van totale titreerbare suur (TTS) en pH, is die chemiese samestelling van wyne met betrekking tot basiese wyn parameters, kleur, fenole en tanniene nie beïnvloed nie. Soortgelyk aan sap is wyn fosfor en geselekteerde ioon samestelling nie geaffekteer nie. Die sensoriese karakteristieke was eenders vir wyne van alle behandelings. Die derde doelwit van die studie was om te bepaal wat die effek wat direkte kontak van kelder afvalwater met druiwekorrels op wyn sensoriese eienskappe het. Hierdie studie het gefokus op die oordrag van afgeure vanaf kelder afvalwater na die wyne sowel as die voorkoms van afgeure as ‘n reaksie op kontak met kelder afvalwater. Wyn kleur en algemene sensoriese eienskappe is nie geaffekteer deur kontak tussen druiwe en kelder afvalwater nie. Kelder afvalwater-geassosieerde afgeure en vlugtige suur was meer duidelik waarneembaar in wyne wat gemaak is van druiwe wat in kontak was met die meer gekonsentreerde afvalwater. Dit mag dus moontlik wees dat afgeure vanaf kelder afvalwater oorgedra word na wyne, of dat sekere afgeure gevorm word as ‘n direkte of indirekte reaksie op kontak met kelder afvalwater. Onder die gegewe toestande oor die twee jaar studie periode het resultate getoon dat besproeiing met verdunde kelder afvalwater nie wingerdprestasie en wyn gehalte aansienlik beïnvloed nie. Die grootste impak wat afvalwater besproeiing gehad het, was om ‘n toename in wyn pH en ‘n tendens tot afname in TTS te veroorsaak. Deur eenvoudig suur by die wyn te voeg kan albei hierdie probleme reg gestel word. Op grond van hierdie bevindings kan besproeiing met verdunde kelder afvalwater moontlik as toekomstige bron vir addisionele wingerdbesproeiing dien. Dit is egter belangrik om te onthou dat die effekte van besproeiing met kelder afvalwater mootlik kumulatief kan wees en dat probleme moontlik eers na etlike jare na vore kan kom. Ander kultivars en veldkondisies mag ook lei tot ander resultate.

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