Thesis (PhD)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Wine is the result of a variety of biochemical reactions and microbial interactions which contribute to the organoleptic properties of wine. Wine aroma and flavour encapsulate the sensory experience of wine and could ultimately determine wine quality and consequently influence consumer acceptance and preference of a product. A thorough understanding of potential factors influencing wine aroma and flavour is therefore needed in order to exploit such factors. The aim of this study was to evaluate the influence of commercial malolactic fermentation (MLF) starter cultures on wine composition, aroma and flavour and the potential impact on consumer preference of experimentally produced red wines. An analytical platform was established to capture the compositional changes induced by different MLF bacteria in experimentally produced red wines. A fast gas chromatography flame ionisation detection (GC-FID) method was developed to determine 39 wine volatile compounds in less than 15 minutes per sample. A 3-fold reduction in analysis time was achieved in comparison to a conventional GC-FID method (40 minutes). Analytes quantified comprise a large boiling point and polarity range which illustrates the robustness of the method. A method was also developed for the direct quantification of carbonyl compounds including diacetyl, acetoin, 2,3-pentanedione and certain aldehydes using headspace solid phase microextraction coupled to gas chromatography mass spectrometry (HS-SPME GC-MS). Both analytical methods showed satisfactory linearity, repeatability and limits of quantification. The contribution of four commercial Oenococcus oeni malolactic fermentation (MLF) starter cultures to the volatile composition, organic acid content and infrared spectral properties of experimentally produced South African red wines, showed significant strain-specific variations in the organic acid profiles, especially for the production of citric acid and lactic acid during MLF. Subsequently, concentrations of compounds related to citric acid metabolism, namely ethyl lactate, acetic acid, diacetyl and acetoin, were influenced accordingly. Bacterial metabolic activity increased the concentration of higher alcohols, fatty acids and esters, with a larger increase observed in ethyl esters compared to acetate esters. A strain-specific tendency to reduce total aldehyde concentrations was found at the completion of MLF, however, further investigation is needed to clarify this observation. Infrared spectral fingerprints were used to characterise the different bacteria and in addition, the prediction of MLF related compounds, diacetyl, acetoin and 2,3-pentanedione, from mid-infrared spectra was explored by partial least squares (PLS) models. Quantitative descriptive analysis (QDA) results depicted significant differences between wines fermented with different starter cultures, in terms of sensory attributes including buttery, fruity, nutty and yoghurt/buttermilk aroma as well as smoothness and mouth-feel attributes. Consumer preference testing results indicate that sensory differences imparted by different MLF bacteria could influence consumer-liking. Preference mapping revealed interesting relationships between sensory attributes and consumer-liking, that can be used for preliminary interpretative purposes. In conclusion, this study illustrated the potential impact of bacterial strains on wine aroma and flavour, resulting sensory properties and consumer preference through an integrative approach combining compositional, spectral, sensory and consumer data. The results presented in this study are of significance to the wine industry since they illustrate and reiterate the potential of different MLF starter cultures as an additional tool to contribute to wine aroma and flavour, and potentially influencing consumer preference and product liking. / AFRIKAANSE OPSOMMING: Wyn is die resultaat van ‘n verskeidenheid biochemiese reaksies en mikrobiologiese interaksies wat tot die organoleptiese eienskappe van die finale produk bydra. Wynaroma en geur vang die sensoriese ervaring van wyn vas en dit kan dus wynkwaliteit bepaal en gevolglik verbruikersaanvaarding asook voorkeur van ‘n produk beïnvloed. Die potensiële faktore wat wynaroma en geur kan beïnvloed moet dus vir hierdie rede deeglik bestudeer word ten einde sulke faktore ten volle te benut. Die doel van hierdie studie was om die invloed van kommersiële applemelksuurgisting (AMG) aanvangskulture op wynsamestelling, die gevolglike aroma en geur eienskappe en die potensiële impak op verbruikersvoorkeure te ondersoek. ‘n Analitiese platform is gevestig om die veranderings in samestelling veroorsaak deur verskillende AMG bakterieë in eksperimenteel bereide rooi wyne vas te vang. ‘n Vinnige gas chromatografiese vlam geïoniseerde deteksie (GC-FID) metode is ontwikkel vir die meting van 39 vlugtige komponente in minder as 15 minute per wynmonster. In vergelyking met ‘n konvensionele GC-FID metode (40 minute) is ’n 3-voudige vermindering in analise tyd behaal. Gekwantifiseerde komponente bestaan uit ‘n wye kookpunt- en polariteitsreeks wat die robustheid van die metode illustreer. ‘n Metode vir die direkte kwantifisering van karboniel komponente, insluitende diasetiel, asetoïen, 2,3-pentanedioon en verskeie aldehiede is ontwikkel met die gebruik van dampfase soliede fase mikroekstraksie gekoppel aan gas chromatografie massa spektrometrie (HS-SPME GC-MS). Albei analitiese metodes besit voldoende lineariteit, herhaalbaarheid en lae deteksie limiete. Die bydrae van vier kommersiële Oenococcus oeni AMG aanvangskulture tot die vlugtige samestelling, organiese suurinhoud en infrarooi spektrale eienskappe van Suid-Afrikaanse rooiwyn het beduidende ras spesifieke variasies in die organiese suur profiele, spesifiek vir die produksie van sitroensuur en melksuur gedurende AMG, vertoon. Gevolglik is die konsentrasies van komponente verwant aan sitroensuur metabolisme, naamlik etiellaktaat, asynsuur, diasetiel en asetoïen, dien ooreenkomstig beïnvloed. Bakteriese metaboliese aktiwiteit het ‘n toename tot gevolg gehad in die hoër alkohole, vetsure en algemene ester konsentrasies met ‘n groter toename in etiel-esters in vergelyking met asetaat-esters. ‘n Ras-spesifieke tendens om die totale aldehiedkonsentrasie te verminder na afloop van AMG, is waargeneem alhoewel verdere ondersoek in hierdie area nodig is. Infrarooi spektrale patrone is gebruik om verskillende bakterieë te karakteriseer asook om die voorspelling van spesifieke AMG verwante komponente soos diasetiel, asetoïen en 2,3-pentanedioon met die gebruik van mid-infrarooi spektrala parsiële kleinste kwadraat verskille (PLS) modelle te ondersoek. Kwantitiewe beskrywende sensoriese analise illustreer beduidende verskille tussen wyne wat gefermenteer is met verskillende aanvangskulture in terme van geure soos botteragtigheid, vrugtigheid, neutagtigheid, joghurt/karringmelkgeur, asook gladheid en mondgevoel eienskappe. Verbruikersvoorkeur resultate illustreer die groot invloed wat sensoriese verskille veroorsaak deur verskillende AMG bakterieë op verbruikersvoorkeure kan hê. Voorkeur kartering het interessante verhoudings tussen sensoriese eienskappe en verbruikersvoorkeure uitgelig. Hierdie studie illustreer die impak van bakteriese rasse op wynaroma en geur en verbruikersvoorkeure deur ‘n geïntegreerde benadering waarin samestellende, spektrale, sensoriese en verbruikersdata gekombineer is. Die resultate van hierdie studie is van belang vir die wynindustrie deurdat dit die potensiële bydrae van verskillende AMG kulture tot wynaroma en geur asook die potensiaal om verbruikersvoorkeure te beïnvloed, illustreer en beklemtoon.

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 (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Introduction: Wine production is a complex biotechnological process aiming at productively coordinating the interactions and outputs of several biological systems, including grapevine and many microorganisms such as wine yeast and wine bacteria. High-throughput data generating tools in the elds of genomics, transcriptomics, proteomics, metabolomics and microbiomics are being applied both locally and globally in order to better understand complex biological systems. As such, the datasets available for analysis and mining include de novo datasets created by collaborators as well as publicly available datasets which one can use to get further insight into the systems under study. In order to model the complexity inherent in and across these datasets it is necessary to develop methods and approaches based on network theory and multivariate data analysis as well as to explore the intersections between these two approaches to data modelling, mining and interpretation. Networks: The traditional reductionist paradigm of analysing single components of a biological system has not provided tools with which to adequately analyse data sets that are attempting to capture systems-level information. Network theory has recently emerged as a new discipline with which to model and analyse complex systems and has arisen from the study of real and often quite large networks derived empirically from the large volumes of data that have collected from communications, internet, nancial and biological systems. This is in stark contrast to previous theoretical approaches to understanding complex systems such as complexity theory, synergetics, chaos theory, self-organised criticality, and fractals which were all sweeping theoretical constructs based on small toy models which proved unable to address the complexity of real world systems. Multivariate Data Analysis: Principle components analysis (PCA) and Partial Least Squares (PLS) regression are commonly used to reduce the dimensionality of a matrix (and amongst matrices in the case of PLS) in which there are a considerable number of potentially related variables. PCA and PLS are variance focused approaches where components are ranked by the amount of variance they each explain. Components are, by de nition, orthogonal to one another and as such, uncorrelated. Aims: This thesis explores the development of Computational Biology tools that are essential to fully exploit the large data sets that are being generated by systems-based approaches in order to gain a better understanding of winerelated organisms such as grapevine (and tobacco as a laboratory-based plant model), plant pathogens, microbes and their interactions. The broad aim of this thesis is therefore to develop computational methods that can be used in an integrated systems-based approach to model and describe di erent aspects of the wine making process from a biological perspective. To achieve this aim, computational methods have been developed and applied in the areas of transcriptomics, phylogenomics, chemiomics and microbiomics. Summary: The primary approaches taken in this thesis have been the use of networks and multivariate data analysis methods to analyse highly dimensional data sets. Furthermore, several of the approaches have started to explore the intersection between networks and multivariate data analysis. This would seem to be a logical progression as both networks and multivariate data analysis are focused on matrix-based data modelling and therefore have many of their roots in linear algebra. / AFRIKAANSE OPSOMMING: Inleiding: Wynproduksie is 'n komplekse biotegnologiese proses wat mik op die produktiewe koördinering van verskeie interaksies en uitsette van verskeie biologiese sisteme. Hierdie sisteme sluit in die wingerd, wat van besondere belang is, asook die wyn gis en wyn bakterieë. Hoë-deurset data generasie word huidiglik beide globaal en plaaslik toegepas in die velde van genomika, transkriptomika, proteomika, metabolomika en mikrobiomika. As sulks is hierdie tipe datastelle beskikbaar vir ontleding, bemyning en verkening. Die datastelle kan de novo gegenereer word, met behulp van medewerkers, of dit kan vanuit die publieke databasisse gewerf word waar sulke datastelle dikwels beskikbaar gemaak word sodat verdere insig verkry kan word met betrekking tot die sisteem onder studie. Die hoë-deurset datastelle onder bespreking bevat 'n hoë mate van inherente kompleksiteit, beide ten opsigte van ditself asook tussen verskeie datastelle. Om ten einde hierdie datastelle en hul inherente kompleksiteit te modelleer is dit nodig om metodes en benaderings te ontwikkel wat gesetel is in netwerk teorie en meerveranderlike statistiek. Verdermeer is dit ook nodig om die kruisings tussen netwerk teorie en meerveranderlike statistiek te verken om sodoende die modellering, bemyning, verkening en interpretasie van data te verbeter. Netwerke: Die tradisionele reduksionistiese paradigma, waarby enkele komponente van 'n biologiese sisteem geontleed word, het tot dusver nie voldoende metodes en gereedskap gelewer waarmee datastelle, wat streef om sisteemvlak informasie te bekom, geontleed kan word nie. Netwerk teorie het na vore gekom as 'n nuwe dissipline wat toegepas kan word vir die model-skepping en ontleding van komplekse sisteme. Dit stem uit die studie van egte, dikwels groot netwerke wat empiries afgelei word uit die groot volumes data wat tans na vore kom vanuit kommunikasie-, internet-, nansiële- en biologiese sisteme. Dit is in skrille kontras met vorige teoretiese benaderings wat gestreef het om komplekse sisteme te verstaan met konsepte soos kompleksiteits teorie, synergetics , chaos teorie, self-georganiseerde kritikaliteit en fraktale. Al die bogeneomde is breë teoretiese konstrukte, gebasseer op relatief kleinskaal modelle, wat nie instaat was om oplossings vir die kompleksiteit van egte-wêreld sisteme te bied nie. Meerveranderlike Data-analise: Hoofkomponente-ontleding (PCA) en Partial Least Squares (PLS) regressie word dikwels gebruik om die dimensionaliteit van 'n matriks (en tussen matrikse in die geval van PLS) te verminder. Hierdie matrikse bevat dikwels 'n aansienlike groot hoeveelheid moontlikverwante veranderlikes. PCA en PLS is variansie gedrewe metodes en behels dat komponente gerang word deur die hoeveelheid variansie wat elke component verduidelik. Komponente is by de nisie ortogonaal ten opsigte van mekaar en as sulks ongekorreleerd. Doelwitte: Hierdie tesis verken die ontwikkeling van verskeie Computational Biology metodes wat noodsaaklik is om ten volle die groot skaal datastelle te benut wat tans deur sisteem-gebasseerde benaderings gegenereer word. Die doel is om beter begrip en kennis van wyn verwante organismes te kry, hierdie organismes sluit in die wingerd (met tabak as laboratorium-gebasseerde plant model), plant patogene en microbes sowel as hulle interaksies. Die breë mikpunt van hierdie tesis is dus om gerekenaardiseerde metodes te ontwikkel wat gebruik kan word in 'n geintergreerde sisteem-gebaseerde benadering tot die modellering en beskrywing van verskillende aspekte van die wynmaak proses vanuit 'n biologiese standpunt. Om die mikpunt te bereik is gerekenaardiseerde metodes ontwikkel en toegepas in die velde van transkriptomika, logenomika, chemiomika en mikrobiomika. Opsomming: Die primêre benadering geneem in hierdie tesis is die gebruik van netwerke en meerveranderlike data-ontleding metodes om hoë-dimensie datastelle te ontleed. Verdermeer, verskeie van die metodes begin om die gemeenskaplike grond tussen netwerke en meerveranderlike data-ontleding te verken. Dit blyk om 'n logiese progressie te wees, aangesien beide netwerke en meerveranderlike data-ontleding gefokus is op matriks-gebaseerde data modellering en dus gewortel is in liniêre algebra.

Thesis (PhD(Agric))-- Stellenbosch University, 2014. / ENGLISH ABSTRACT: Brandy producers in South Africa can produce three categories of brandy, and within those categories, varies styles in order to cater for diverse consumer needs. Thus they have divergent opinions as to what comprises a good base wine and distillate for their individual end-product style in mind. Due to the fact that brandy is the largest locally produced spirit in South Africa, it is crucial to investigate the factors that influence the production of brandy as better understanding and control of these processes leads to the production of a brandy that is more consistent in quality and in style. The factors known to influence the quality of brandy are the grape cultivar, vinification techniques, distillation and the oak maturation process; however limited research has been done on the factors contributing to the style of brandies. Understanding and identifying the factors that contribute to the style of brandy, will allow for better control and manipulation of the process to ultimately produce a spirit product with a desired style classification. In order to do so, one must distinguish as to what differentiates (chemical and sensory analyses) the two most divergent styles of brandy (in this study referred to as class one and class five three-year barrel matured brandy). Over 104 three-year old barrel matured brandy samples were collected and analysed using GC-FID and HPLC analysis over a three year period. Of these samples, only 7 were classified as a class one three-year barrel matured brandy and 23 were found to be classified as class five three-year barrel matured brandy. The results obtained showed that the class one samples were found to contain a higher concentration of total higher alcohols, higher ratio of higher alcohols vs. esters, higher alcohols vs. acids and aldehydes vs. esters, but a lower level of total esters and acids. The class five samples contained a higher concentration of total esters, furans, lactones and acids, but a lower level of total higher alcohols and ratio of higher alcohols vs. esters. The factors investigated that possibly influence the style of brandy were: the base wine prior to distillation, yeast strain, fermentation temperature, condenser water temperature and barrel age. Results of the base wine analysed prior to distillation show that those samples that scored a sensory score of one could produce a class one three-year barrel matured brandy, and base wines that scored a sensory score of five would yield a class five three-year barrel matured brandy. Alchemy 1 yeast was found to produce elevated levels of total esters and could by default result in a class five three-year barrel matured brandy. Similarly if the wine was fermented at 24⁰C it would result in the increased production of higher alcohols and total acids and in-turn could yield a class one three-year barrel matured brandy. The results obtained show that the different condenser water temperatures do not necessarily influence the concentration of esters and higher alcohols. However, if a lower condenser water temperature (8⁰C) was employed, it could yield a class one barrel matured brandy sample. These distillates are associated with a higher concentration of total higher alcohols and even though they may be positively associated with higher levels of esters, the esters could be removed during the head fraction in the distillation process. The results also show that during the maturation process that the use of 18 year old barrels for the maturation of three-year barrel matured brandy samples could possibly produce a class one three-year barrel matured brandy and the use of new barrels could yield a class five three-year barrel matured brandy sample. It is recommended that analysing the base wine prior to distillation using GC-FID to identify the chemical composition is the most important step in determining the outcome of the style of brandy. / AFRIKAANSE OPSOMMING: Brandewyn produsente in Suid-Afrika kan drie kategorieë brandewyn produseer, en binne hierdie kategorieë sekere style om voorsiening te maak vir die diverse behoeftes van die mark, en dus het hulle uiteenlopende menings oor wat 'n goeie basis wyn en distillaat behels vir hul individuele eindproduk se spesifieke styl. As gevolg van die feit dat brandewyn die belangrikste spiritus produk is wat deur die Suid-Afrikaanse bevolking verbruik word, is dit noodsaaklik dat dié faktore wat die produksie van brandewyn beïenvloed ondersoek word om 'n beter begrip van, en beheer oor hierdie prosesse verkry kan. Dit sal lei tot die produksie van 'n brandewyn van konstante in qualitiet en in style. Die faktore wat bekend is om die gehalte van brandewyn te beïnvloed, is die druifkultivar, wynbereidingstegnieke, distillasie en die hout verouderings proses, maar daar is beperkte navorsing oor die faktore wat bydra tot die styl van n brandewyn. Om die faktore wat bydra tot die spesifieke styl van n brandewyn te identifiseer en te verstaan, kan dit lei tot 'n beter beheer en manipulasie van die prosess om uiteindelik n brandewyn met 'n gewenste styl te produseer. Ten einde dit te kan doen, moet 'n mens kan bepaal wat die twee uiterste style van brandewyn (wat in die studie sal bekend wees as klas een en klas vyf drie-jaar vat veroude brandewyn) van mekaar onderskei (chemiese en sensoriese analise). Meer as 104 drie-jaar vat verouderde brandewyn monsters is ingesamel en ontleed met behulp van GC-FID en HPLC analyliese oor 'n tydperk van drie jaar. Van die 104 monsters, is slegs 7 as 'n klas een drie-jaar vat verourde brandewyn geklassifiseer en 23 as klas vyf drie-jaar vat verourde brandewyn geklassifiseer. Die resultate het getoon dat die klas een monsters 'n hoër konsentrasie van die totale hoër alkohole, 'n hoër verhouding van hoër alkohole teenoor esters het, hoër alkohole teenoor sure en aldehiede teenoor esters, maar 'n laer vlak van totale esters en sure. Die klas vyf monsters het 'n hoër konsentrasie van die totale esters, furanen, laktone en sure, maar 'n laer vlak van totale hoer alkohol en verhouding van hoër alkohole teenoor esters. Die faktore wat ondersoek was wat moontlike ‘n invloed op die styl van brandewyn kon hê, was die basis wyn voor distillasie, die gisras, die fermentasie temperatuur, die kondensor water temperatuur en die vat ouderdom. Die ontledings van die basis wyn voor distillasie het gewys dat die monsters wat 'n sensorise telling van een behaal het, moontlik kan lei tot 'n klas een drie-jaar vat verouderde brandewyn, en basis wyne wat 'n sensorise telling van vyf gekry het weer ‘n klas vyf drie-jaar vat verouderde brandewyn sou oplewer. Alchemie 1 gis ras het ‘n verhoogde vlak van totale esters geproduseer wat tot 'n klas vyf drie-jaar vat verouder brandewyn gelei het As die basis wyn gefermenteer is teen 24⁰C sal dit lei tot die verhoogde produksie van hoër alkohole en totale sure wat 'n klas een drie-jaar vat verouderde brandewyn kan oplewer. Die resultate verkry, toon dat die verskillende kondensor water temperature nie noodwendig die konsentrasie van esters en hoër alkohole beïnvloed het nie, maar dit wil voorkom dat indien 'n laer kondensor water temperatuur (8⁰C) gebruik is, kan dit 'n klas een vat verouderde brandewyn oplewer. Dit hou verband met 'n hoër konsentrasie van die totale hoër alkohole en selfs al word die distillate positief geassosieer met hoër vlakke van esters, kan die esters verwyder word tydens die verwydering van die voorloop in die distillasie proses. Die resultate toon ook dat die gebruik van 18 jaar oue vate vir die veroudering van brandewyn moontlik 'n klas een drie-jaar vat verouderde brandewyn kan oplewer en die gebruik van nuwe vate kan 'n klas vyf drie-jaar vat verouderde brandewyn lewer. Dit word egter aanbeveel dat die basis wyn voor distillasie met GC-FID chemies ontleed word om die samestelling van die wyn te identifiseer Die resultate bly die beste aanduiding van die styl van die brandewyn.

Thesis (PhD)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Brandy has a high alcohol content of at least 36% alcohol by volume and a complex volatile chemical structure. Due to this, the sensory evaluation of brandy is challenging to execute and must be carefully managed to avoid panellists’ sensory and mental fatigue. Although rapid sensory profiling methods, such as projective mapping (PM), are commonly used for fast moving consumer goods, it was unclear whether these methods could accommodate the difficulties associated with brandy evaluation. The primary objective of this study was to validate PM, also called Napping®, as a reliable tool for brandy evaluation. Two variations of PM were tested: global napping (GN) where products are evaluated based on overall sensory perception, and partial napping (PN) where the product perception is broken down in to separate sensory modalities, in this case appearance, aroma and in-mouth sensations. Several practical aspects of PM were investigated in order to optimise the method for brandy evaluation and to gain understanding into practical methodological aspects that have not been fully understood at the onset of this study. The results showed that both GN and PN delivered reliable results, but that PN was more reproducible and better suited to larger sample sets (10 products). The concept of in-mouth sensations was found to be ineffective in extracting useful information on mouthfeel differences in the product set and that retronasal flavour should be separated from basic tastes and mouthfeel. A verbal instruction to the panellists was sufficient to obtain reliable information on mouthfeel differences; it was not necessary to use black glasses or nose-clips to eliminate the influence of appearance and flavour perception. In response to the insufficient reproducibility of GN, the sorting method was tested and validated as a suitable screening method that delivered reliable product maps of larger sample sets (10 products) in one replication, compared to GN which has to be replicated several times. A recent hypothesis states that the elongated horizontal dimension of a rectangular PM tasting sheet could be used to elicit more prominent product differences, while tasting sheets without an elongated axis, such as circles or squares, could be used to bring forth more subtle differences. This hypothesis was challenged by testing rectangular, square and round tasting sheet shapes. Although differences were observed between the shapes in terms of product configuration and panellist performance, the practical value of using different tasting sheet shapes to obtain specific information could not be established. The impact of high alcohol content and product complexity on panellist performance in PM was tested and it was concluded that product complexity did influence the panellists’ performance, particularly for high alcohol products. It was also found that prior knowledge of a products’ high alcohol content influenced the panellists’ descriptive language to include more alcohol-related terms. A new panellist performance measure was developed, namely the Relative Performance Index (RPI). This measure can be used to monitor trained panellists’ performance in the PM task more effectively to thereby ensure reliable results. The outcomes of this study extended the brandy sensory evaluation toolbox. Practical measures were identified that can be used to overcome the challenges associated with the sensory evaluation of complex high alcohol products. / AFRIKAANSE OPSOMMING: Met ‘n alkohol inhoud van mintens 36% alkohol per volume en ‘n komplekse vlugtige chemise samestelling is brandewyn ‘n uitdagende produk om sensories te evalueer. Maatreëls moet in plek gestel word om te verhoed dat sensoriese paneellede nie uitgeput raak nie om sodoende betroubare resultate te verseker. Hoewel vinnige sensoriese toetsmetodes, soos projeksiekartering (PM) en sortering, gereeld gebruik word vir die evaluasie van vining bewegende verbruikersgoedere was dit weens die voorafgenoemde redes onduidelik of die metodes geskik sou wees vir brandewynevaluasie. Die oorhoofse doelwit van hierdie projek was om PM, ook genoem Napping®, te valideer as ‘n geskikte metode vir die sensoriese evaluering van brandewyn. Twee variasies van PM is ondersoek: oorhoofse kartering (GN) waar die produkte op grond van ‘n algehele sensoriese waarneming evalueer word, en gedeeltelike kartering (PN), waar die verskillende sensoriese waarnemingsmodaliteite afsonderlik gemeet word. In hierdie geval is voorkoms, aroma en algehele binne-mondse ervaring gemeet. Verskeie aspekte van GN en PN is ondersoek ten einde die metodes vir brandewynevaluasie te optimiseer asook lig te werp op praktiese metodologiese aspekte waaroor daar nog onsekerheid was by die aanvang van hierdie studie. Die resultate het getoon dat beide GN en PN betroubare inligting oor brandewyn kan oplewer, maar dat PN meer herhaalbaar en beter geskik is vir groter produkstelle (10 produkte). Dit is egter bevind dat die konsep van algehele binne-mondse waarneming nie doeltreffend was om mondgevoelverskille tussen produkte uit te lig nie. Smaak- en mondgevoelwaarnemings moet liefs onderskei word van binne-mondse geurwaarneming. Dis bevind dat dit voldoende is om hierdie onderskeid deur skriftelike instruksies tot die paneel te bewerkstelling en dat die gebruik van swart glase en neusknippies nie nodig was om betekenisvolle inligting oor mondgevoel te bekom nie. Na aanleiding van die swak herhaalbaarheid van GN, is sortering getoets en gevalideer as ‘n geskikte siftingsmetode wat betroubare produkkaarte van groot produkstelle (10 produkte) kan oplewer met slegs een evaluasie, teenoor GN waarvoor meer as een sessie benodig word. ‘n Onlangse hipotese stel dat die langer horisontale dimensie van reghoekige PM proekaarte gebruik kan word om meer prominente verskille tussen produkte uit te wys, terwyl proekaarte sonder verlengde asse, soos sirkels en vierkante, gebruik kan word om meer subtiele verskille uit te wys. Hierdie hipotese is bevraagteken deur reghoekige, vierkantige en ronde PM proekaarte teenoor mekaar te toets. Hoewel die resultate vanaf die verskillende proekaarte effens verskil het in terme van produk konfigurasie en die paneel se taakverrigting, kon die praktiese waarde van die gebruik van produkkaarte met verskillende vorms om spesifieke inligting te bekom nie bevestig word nie. Die impak van hoë alkohol en produkkompleksiteit op paneellede se PM taakverrigting is ondersoek. Dit is bevind dat produkkompleksiteit wel hul taakuitvoering beinvloed, en dat hierdie invloed groter is vir hoë alkohol produkte. Dit is ook gevind dat paneellede se beskrywende taalgebruik beïnvloed om meer alkoholverwante woorde in te sluit word indien hul kennis dra van ‘n produk se hoë alkohol inhoud. ‘n Nuwe vaardigheidsberaming vir die meet van paneellede se taakverrigting is ontwikkel, naamlik die Relatiewe Taakverrigtingsindeks (RTW). Dit kan gebruik word om paneellede se vaardigheid in PM meer doeltreffend te monitor en sodoende betroubare resultate te verseker. Die uitkomste van die studie het die poel beskikbare metodes vir die sensoriese evaluering van brandewyn suksesvol verbreed en praktiese maatreëls is geidentifiseer om die uitdagings van ‘n komplekse hoë alkohol produkte te oorkom.

Thesis (MSc (Wine Biotechnology))--University of Stellenbosch, 2009. / Saccharomyces cerevisiae has been used for millennia for the leavening of dough and in the production of alcoholic beverages such as beer and wine. More recently, it is being used as cell factories for the production of important pharmaceutical products. S. cerevisiae has also been extensively used as a model organism for studying many genetic and biochemical processes within the eukaryotic cell. Since the completion of a yeast genome sequence, many functional analysis projects have emerged with the aim of elucidating the functions of the unidentified genes revealed by the genome sequence. One of the most relevant approaches consisted in the construction of a collection of mutants deficient in all single genes, either in a haploid background for non-essential genes, or as heterozygous diploids for essential genes. This collection of strains can be subjected to phenotypic screens that might reveal the function of unknown genes or add to our understanding of already annotated genes. While this approach is promising, it also bears some limitations. For instance, many mutants have no overt phenotypes and some phenotypes do not obviously showcase the function of the encoded protein. In this study, S. cerevisiae strains with single deletions of genes involved in pyruvate metabolism were selected from the Euroscarf deletion library. Pyruvate is a central intermediate of glycolysis, and pyruvate metabolism largely defines the general distribution of carbon flux in the cell. These mutants were screened for modified fermentation kinetics or modified carbon flux under wine fermentative conditions, an environment that had not been previously used for the analysis of these mutants. A strain disrupted in the PDA1 gene, which encodes the E1α subunit of the pyruvate dehydrogenase showed a significant change in phenotype when grown in wine fermentative conditions. In particular, the mutant displayed a prolonged lag phase, but upon entering exponential growth, fermented significantly faster than the wild type strain and completed alcoholic fermentation in a shorter period of time. This phenotype could be of significant industrial interest. The mutant phenotype was further investigated through disruption of the gene in the same as well as in different genetic backgrounds, and through complementation of the PDA1 deletion with a plasmid-born wild type copy. The data show that the PDA1 gene disruption is not solely responsible for the observed phenotypes under wine fermentative conditions. We therefore propose that secondary mutations have contributed to the mutant phenotype. This study shows that phenotypes attributed to a specific gene in mutants of the Euroscarf library should always be confirmed before performing consequent experiments and drawing significant conclusions.

Thesis (MScAgric (Viticulture and Oenology. Wine Biotechnology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: The Vitis vinifera L. carotenoid cleavage dioxygenase 1 gene (VvCCD1) is a member of a structurally conserved gene family encoding enzymes that cleave multiple carotenoid substrates to form apocarotenoids. Carotenoid pigments are synthesised in the chloroplast where they are primarily involved in light harvesting and photo-protection during photosynthesis while apocarotenoids fulfill diverse roles that range from pollinator attractants to phytohormones. CCD1 cleaves carotenoids at specific double bond sites producing volatile apocarotenoids. These CCD1-derived apocarotenoids typically possess a fruity and floral aroma, thus making them desirable targets for metabolic engineering. CCD1 orthologues are highly homologous and have been isolated and characterised from a number of plant species, including Arabidopsis, tomato, rose, petunia, and grapevine. VvCCD1 is localised to the cytosol and has been shown in vitro to cleave zeaxanthin and lutein resulting in 3-hydroxy-β-ionone. Expression of VvCCD1 increases during berry ripening, peaking at véraison. Due to the impact that VvCCD1 potentially has on the flavour and aroma of grape berries and therefore wine, this study aimed to characterise the specific enzyme action as well as the biological role that this enzyme plays in grapevine. Expression of VvCCD1 in carotenoid-accumulating Escherichia coli strains demonstrated cleavage of β-carotene at the 9,10 (9’,10’) position forming β-ionone; and lycopene at the 5,6 (5’,6’) and 9,10 (9’,10’) position, forming 6-methyl-5-hepten-2-one and pseudoionone, respectively. A transgenic grapevine population with modified VvCCD1 expression was generated and genetically and metabolically characterised. The transgenic population consisted of lines in which VvCCD1 was either overexpressed or silenced. Expression analysis of stable transformants showed a 12-fold range of VvCCD1 expression relative to the wild-type. HPLC analysis of the photosynthetic pigment content of the transgenic population necessitated the development and optimisation of a method for the extraction of pigments, specifically from grapevine. A number of parameters were identified and optimised, resulting in a method that provides accurate quantification of photosynthetic pigments from grape berries and leaves. Absolute quantification of the following major photosynthetic pigments present in grapevine is now possible: chlorophyll a, chlorophyll b, lutein, -carotene, zeaxanthin, antheraxanthin, violaxanthin and neoxanthin. Data suggest that various levels of molecular control regulate carotenoid cleavage and apocarotenoid biosynthesis. The majority of lines stably transformed with a VvCCD1 overexpression cassette exhibit post-transcriptional gene silencing. Expression analysis in these lines demonstrated that, despite the additional contribution of transgene-derived VvCCD1 transcripts, the total VvCCD1 transcript levels were not significantly higher than in wild-type lines. In lines where transgenic manipulation of VvCCD1 expression was successful, subsequent analysis of carotenoids and apocarotenoids in leaf tissue showed no correlation between the measured metabolites and gene expression. The in planta action of VvCCD1 is presumably distinct from the observed in vitro activity due to the strict compartmentalisation required in photosynthetic leaf tissue preventing access of cytosolic VvCCD1 to the chloroplastic carotenoids. Future studies on reproductive organs (grape berries) from the transgenic lines generated in this study will be of great importance in further elucidation of the in planta function of VvCCD1. / AFRIKAANSE OPSOMMING: Die Vitis vinifera L. “carotenoid cleavage dioxygenase” 1 geen (VvCCD1) behoort aan ‘n geenfamilie wat struktureel gekonserveerd is en kodeer vir ensieme wat verskeie karotenoïed substrate afbreek om apokarotenoïede te vorm. Karotenoïed pigmente word in die chloroplaste gesintetiseer waar hulle primêr betrokke is by lig-insameling, sowel as beskerming tydens fotosintese, terwyl apokarotenoïede diverse funksies in die plant verrig wat strek van aantrekking van stuifmeelverspreiders tot phytohormone. CCD1 breek karotenoïede by spesifieke dubbelbindingsetels af om vlugtige apokarotenoïede te vorm. Die apokarotenoïede wat van CCD1 afkomstig is besit tipies vrugtige en blomagtige aromas wat hul gesogte teikens maak vir metaboliese manipulering. CCD1 ortoloë is hoogs homoloog en is al geїsoleer en gekarakteriseer vanuit ‘n verskeidenheid plantspesies wat Arabidopsis, tamatie, roos, petunia en wingerd insluit. VvCCD1 is in die sitosol gelokaliseer en dit is vantevore gewys dat dit beide zeaxanthin en lutein in vitro kan afbreek om 3-hidroksi-b-ionoon te vorm. Die uitdrukking van VvCCD1 vermeerder tydens korrel rypwording en bereik ‘n maksimum tydens véraison. Weens die potensieële invloed vanVvCCD1 op die geur en aroma van druiwe, en dus wyn, is hierdie studie gerig op die karakterisering van die spesifieke ensiematiese aksie, sowel as die biologiese rol van hierdie ensiem in wingerd. Uitdrukking van VvCCD1 in Escherichia coli rasse wat karotenoïede versamel het getoon dat β-karoteen by die 9,10 (9’,10’) posisie afgebreek word om β-ionoon te vorm, en likopeen by die 5,6 (5’,6’) en 9,10 (9’,10’) posisie om onderskeidelik 6-metiel-5-hepteen-2-oon en pseudo-ionoon te vorm. ‘n Transgeniese wingerd populasie is gegenereer met gewysigde VvCCD1 uitdrukking en is geneties en metabolies gekarakteriseer. Die transgeniese populasie het bestaan uit lyne waar VvCCD1 óf ooruitgedruk óf afgereguleer is. Uitdrukkings analise van die stabiele transformante het ‘n 12-voudige reeks van VvCCD1 uitdrukking getoon, relatief tot die wilde tipe. HPLC analise van die fotosintetiese-pigment inhoud van die transgeniese populasie het die ontwikkeling en optimisering van ‘n wingerd-spesifieke metode vir die ekstraksie van pigmente genoodsaak. ‘n Aantal parameters is geïdentifiseer en geoptimiseer, en het gelei tot ‘n metode wat akkurate kwantifisering van fotosintetiese pigmente in druiwe en wingerdblare kan lewer. Absolute kwantifisering van die volgende belangrike fotosintetiese pigmente aanwesig in wingerd is nou moontlik: chlorophyll a, chlorophyll b, lutein, -karoteen, zeaxantien, anteraxantien, violaxantien en neoxantien. Data dui aan dat verskeie vlakke van molekulêre beheer die afbreking van karoteen en die biosintese van apokarotenoïede reguleer. Die meerderheid van die lyne wat stabiel getransformeer is met ‘n VvCCD1 ooruitdrukkingskasset het na-transkripsioneleafregulering van die geen getoon. Uitdrukking analise van die lyne het gewys dat ten spyte van die addisionele transgeniese VvCCD1 transkripte, die totale VvCCD1 transkripvlakke nie beduidend hoër was as dié van die wilde-tipe lyne nie. In die lyne waar transgeniese manipulasie van VvCCD1 uitdrukking wel suksesvol was, het verdere analise van die karotenoïed en apokarotenoïed vlakke in blaarweefsel geen korrelasie getoon tussen die metaboliete en VvCCD1 uitdrukking nie. Die in planta aktiwiteit van VvCCD1 is vermoedelik anders as die in vitro aktiwiteit weens die streng kompartementalisering benodig in fotosintetiese blaarweefsel, wat verhoed dat die sitosoliese VvCCD1 toegang het tot die chloroplastiese karotenoïede. Toekomstige bestudering van die reproduktiewe organe (druiwe) van die transgeniese lyne wat in hierdie studie gegenereer is sal belangrik wees in die verdere verduideliking van die in planta funksie van VvCCD1.

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: The yeast Saccharomyces cerevisiae is able to sense and respond to changes in its immediate environment. Information regarding the nutritional status of the extracellular environment is sensed by membrane receptor systems and relayed through signalling pathways to the nuclear interior, affecting the transcription of specific genes., Transcription factors, which function downstream of these signal transduction pathways, have to be transported into the nucleus after synthesis in the cytoplasm in order to regulate transcriptional events. Transport into the nucleus occurs in a tightly regulated manner at the nuclear pore complex, which is located in the nuclear membrane, and requires the recognition of transport signal sequences, which are present in the proteins that are to be transported. Signalling pathways control the nuclear accessibility of transcriptional regulators by modifying their respective signal sequences. In response to a limited availability of carbon or nitrogen, cells are able to change their morphology from a unicellular ovoid form to elongated cells attached to each other. This morphological change is associated with daughter cells that remain attached to their respective mother cells following unipolar budding, thus forming filamentous structures referred to as pseudohyphae. The regulation of the development of pseudohyphae is correlated with other physiological processes, such as starch degradation and the invasion of agar-containing media. Mss11p performs a central role in the regulation of the genes required for these processes and it has been shown to specifically regulate the expression of FL011, which encodes a cell surface protein critical for pseudohyphal development, and STA2, which encodes an extracellular glucoamylase functioning in the degradation of starch. The aim of this study was to characterise the functioning of Mss11p. Overexpression analysis indicates that Mss11p functions as an inducer of invasive growth, cell elongation and flocculation. Furthermore, MSS11 deletion improves biomass formation and suppresses the growth defect of yeast from a L:1278b genetic background transformed with the RAS2val19 allele on non-fermentable carbon sources. Biochemical analysis shows that Mss11p is a nuclear protein of approximately 97 kDa in apparent size that is maintained at relatively low levels in yeast. Finally, the data suggest a model in which Mss11p functions as a mediator of the transcriptional regulation of various genes. / AFRIKAANSE OPSOMMING: Die gis Saccharomyces cerevisiae is in staat om veranderinge in sy onmiddelike omgewing waar te neem en daarop te reageer. Inligting betreffende die beskikbaarheid van voedingstowwe in die omgewing word vanaf membraan reseptorsisteme deur middel van seintransduksiekaskades na die nukleus herlei, waar die transkripsie van spesifieke gene beïnvloed word. Transkripsie faktore wat stroom af van hierdie seintransduksie funksioneer, moet na die nukleus vervoer word na vervaardiging in die sitoplasma, om sodoende transkripsionele gebeurtenisse te reguleer. Die vervoer van faktore na die binnekant van die nukleus vind onder streng regulering plaas by die nukleêre porie kompleks, wat in die nukleêre membraan gesitueer is. Vervoer vind plaas deur middel van die herkenning van nukleêre lokaliseringsekwense wat in die proteïene wat vervoer word, teenwoordig is. Seintransduksiekaskades beheer die beskikbaarheid van proteïene tot die nukleus deur hulonderskeidelike nukleêre lokaliseringsekwense te modifiseer. Selle is in staat om hul morfologie te verander van 'n eensellige eliptiese vorm tot verlengde selle wat aan mekaar geheg bly in reaksie op die beperkende beskikbaarheid van koolstof of stikstof bronne. Hierdie morfologiese verandering word geassosieer met dogterselle wat ná monopolêre botselvorming aan hul moederselle geheg bly, en dus filamentagtige strukture vorm wat pseudohifes genoem word. Die regulering van die ontwikkeling van pseudohifes word gekorreleer met ander fisiologiese prosesse, soos styselafbraak en die penetrerende groei van selle op agar-bevattende media. Mss11p vervul 'n sentrale rol in die regulering van gene wat vir hierdie prosesse benodig word en reguleer die uitdrukking van FL011, wat kodeer vir 'n selwandproteïen wat krities is vir die ontwikkeling van pseudohifes, en STA2, wat kodeer vir 'n ekstrasellulêre glukoamilase wat vir die afbraak van stysel benodig word. Die doel van hierdie studie was om Mss11p-funksie te karakteriseer. Deur middel van oorproduksie is Mss11p as die induseerder van penetrerende groei, selverlenging en flokkulasie geïdentifiseer. Verder is bevind dat MSS11-delesie lei tot verhoogde biomassa formasie, en dat dieselfde delesie lei tot 'n oorkoming van 'n groeidefek van gis van die 2:1278b genetiese agtergrond wat met die RAS2val19aleel op nie-fermenteerbare koolstofbronne getransformeer is. Biochemiese analise dui daarop dat Mss11p 'n nukluêre proteïen is van ongeveer 97 kDa in oënskynlike grootte, wat teen lae vlakke in gis onderhou word. Die data stel 'n model voor waarin Mss11p as bemiddelaar optree vir die transkripsionele regulering van verskeie gene.

Thesis (MScAgric)--Stellenbosch University, 1946. / No Abstract Available

Thesis (MScAgric)--Stellenbosch University, 1954. / No Abstract Available