The unique flavor of high quality Swiss cheese is difficult to reproduce in commercial market cheese. Swiss cheese flavor has never been duplicated or thoroughly understood. New techniques and advances in flavor research have enabled better definition and understanding of food flavors. Therefore, it was desirable to make a detailed investigation of Swiss cheese flavor. Neutral volatile flavor compounds were isolated from Swiss cheese fat by low-temperature low-pressure distillation. The compounds were separated by temperature programmed gas chromatography. Direct analysis of cheese fat and whole cheese from four domestic and two imported good flavored cheeses by gas entrainment and on-column trapping provided a further means of isolation of volatile flavor compounds in Swiss cheese. Gas chromatography in conjunction with rapid scan mass spectrometry and relative retention time data were used to identify compounds. Compounds positively identified by the distillation and on-column trapping techniques were as follows: methanol, ethanol, 1-propanol, 1-butanol, 2-pentanol, trans-2-hexene-1-ol, 2-phenylethanol, acetaldehyde, 2-methyl propanal, 2-methyl butyraldehyde, benzaldehyde, phenylacetaldehyde, acetone, butanone, 2-pentanone, 2-hexanone, 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, 2-pentadecanone, hexane, octane, 1-octene, nonane, 1-nonene, dodecane, pentadecane, toluene, α-pinene, methyl acetate, methyl hexanoate, methyl octanoate, methyl decanoate, ethyl propionate, ethyl butanoate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, ethyl dodecanoate, butyl acetate, 3-methyl butyl acetate, γ-valerolactone, γ-dodecalactone, δ-octalactone, δ-decalactone, δ-dodecalactone, dimethyl sulfide, diacetyl, benzothiazole, o-dichlorobenzene, 1, 2, 4-trichlorobenzene, di-isobutyl adipate, and chloroform. Compounds tentatively identified include an aromatic hydrocarbon, pinane, α-fenchene, ethyl benzene, a di-methyl benzene, methyl benzoate, 2-phenyl-2-methyl butane, 5-methyl-5-ethyl decane, 3-methyl butyl octanoate, 2, 5-dimethyl tetra decane, methyl vinyl ether and 2-methyl propenal. The concentration of selected volatile compounds identified by the on-column trapping technique were determined by relating their peak heights to known quantities of compound. Average concentrations calculated from the mean values for all the six cheeses and expressed in parts per million were as follows: dimethyl sulfide. 0.107; diacetyl, 0.8; acetaldehyde, 1.4; acetone, 1.6; butanone, 0.3; 2-methyl butyraldehyde, 0.42; 2-pentanone, 0.98; 2-heptanone, 0.45; ethanol, 16.3; 2-butanol, 0.3; 1-propanol, 2.9; 1-butanol, 0.7; methyl hexanoate, 1.5; and ethyl butanoate, 0.6. Liquid-liquid partition chromatography and gas chromatography were utilized to determine quantitatively the major free, fatty acids in the six Swiss cheeses. 2-Methyl butyric acid was detected in all cheeses and varied from 9.0 to 100.0 mg/kg cheese. The other isomeric acid, 3-methyl butyric, was detected in only two cheeses. Formic acid was detected in only one cheese. No n-valeric or 2-methyl propionic acids were detected. A synthetic Swiss cheese flavor was prepared utilizing the data obtained in this investigation and that available in the literature for free amino acids. A satisfactory reproduction of Swiss cheese flavor could be achieved only if the mixture contained free fatty acids, volatile constituents, and free amino acids and was adjusted to the pH of natural cheese. / Graduation date: 1966

The flora of Aotearoa/ New Zealand has evolved in association with birds and lizards as the dominant frugivores and seed dispersers. There is a wide range of ripe fruit colours within the native fleshy-fruited plants spanning the visible light spectrum from red to violet, with the notable exception of green. The evolution of fleshy-fruit and fruiting related trait, may be a result of the selection pressures exerted by different frugivore guilds. This study was conducted to ascertain if endemic diurnal lizards, Oligosoma species (Scincidae: Lacertilia), display features associated with visual based foraging, colour sensitivity and colour preferences, which are necessary conditions to infer a co-evolutionary mutualism between fleshy-fruited plant species and lizards as seed dispersers, which may have influenced the evolution of fruit colour. Many lizards have exceptional colour vision, with the ability to see a wide range of the visual light spectrum from short wave ultra violet to long wave red. They are able to discriminate all aspects of colour: hue, brightness and saturation. Fruit colour within Coprosma (Rubiaceae), is extremely variable, between and within species. The study of fruit colour preferences within this genus and particularly within species with polymorphic fruit colour provides a valuable comparison of frugivore preference to fruit with little inter and intraspecific variation, therefore minimising potentially confounding factors due to phylogeny. Fruit-colour choice experiments were conducted offering fruit from two colour categories based on postulated frugivore preferences; red and red orange fruit has been associated with avian frugivores whilst white and pale fruit has been associated with lizard frugivory in New Zealand. Experiments were conducted both ex-situ, in environmentally controlled laboratories and in-situ at Macraes Flat, Otago. Pilot trials indicated that the background colour on which fruit were presented was important in fruit choice and consequentially, all fruit were offered on a background which provided contrast to both fruit colour categories. The laboratory trials showed some weak evidence for a preference of white and pale blue fruit however, in-situ trials showed a strong preference for white over red fruit. Field studies were conducted to ascertain the composition of fleshy-fruit in the diets of lizards and the results were consistent with those expected for a generalist omnivore; many of the small fruits available to lizards were consumed however, the results indicated that plant abundance does not adequately explain fruit consumption at this field site. A preference index showed that white and pale fruited plants, Melicytus alpinus and Coprosma spp., were preferred over more abundant orange and red fruited plants. Fruit odour was investigated to determine if fruit choice was mediated primarily by visual cues as opposed to odour cues. Fruit choice trials with the fruit concealed from view indicated that fruit choice was based primarily on visual cues in Oligosoma skinks. It is concluded that lizards demonstrate the necessary conditions to infer that as frugivores, they may have influenced the evolution of fruit colour and that within the open habitats of Aotearoa/ New Zealand, the shrubs, particularly the divaricate shrubs may have provided sufficient environmental conditions to establish a mutualism between plants and lizards resulting in the evolution of small, white and other low chroma fruits.

Hops (Humulus lupulus L.) are primarily used to provide specific characteristics to beer, such as bitterness, aroma, flavor, and microbial stability. The chemical composition of hops, relative to how they are used during the brewing process, dictates the expression of these characteristics. Of the raw ingredients that go into making beer, hops are perhaps the most costly. Considerable resources are required to grow quality hops, and therefore, brewers and hop growers alike have a common goal of obtaining the highest quality hops possible. However, quality can be a relative term. While it is commonly agreed upon that high brewing values, such as α-acids and essential oil content, and robust structural integrity are indicators of quality hops, there are many opinions of the ideal aroma. Changes in the chemical composition of hops during plant maturation are a dynamic process requiring a comprehensive, in-depth chemical and sensory analysis in order to maximize the characteristics of interest to brewers. The complex aroma chemistry associated with hops in beer has been a confounding variable for the practical brewer, and a deeper understanding of hop aroma development during cultivation is needed. The effect of harvest date, location, and cultivar on key chemical components of Willamette and Cascade hops was investigated for the 2010 and 2011 growing seasons. Hops were harvested at 3 time points (Early, Typical, and Late), within a 3-week interval from 2 different farms in the Willamette Valley, Oregon. A split-plot experimental design for each cultivar was used; each farm represented a main plot and harvest years were designated as subplots. American Society of Brewing Chemist standard methods of analysis were used to measure moisture content, hop acids and their homologs, Hop Storage Index, total essential oil content and volatile profile by GC-FID. Additionally, difference testing, descriptive analysis, and consumer acceptance testing was conducted using beers brewed with either Typical or Late harvested Cascade hops from the 2010 harvest year. The response of analytes was dependent on the cultivar being examined, its location within the Willamette Valley, as well as days until harvest. Hop acids did not change appreciably during plant maturation for the period examined, while hop oil content increased hyperbolically to a plateau as the hops aged on the bine. Increases in oil quantity were strongly correlated (r > 0.90) with increases in α-pinene, β-pinene, myrcene, limonene, methyl heptanoate, and linalool concentrations. For Cascade, α-pinene, β-pinene, myrcene, limonene, ρ-cymene, caryophyllene, E, β-farnesene, and humulene all increased from Early to Typical points but no increase was observed between the Typical and Late time point. Linalool and methyl heptanoate increased between each time point while citral and humulene epoxide differed between Early harvest and Late harvest, but not between Early and Typical or Late and Typical harvests. For Willamette hops, α-pinene, β-pinene, myrcene, limonene, ρ-cymene, and linalool all increased between each time point. Caryophyllene, E β-farnesene, humulene, farnesol and citral all increased from Early harvest to Typical harvest but no difference was observed between Typical and Late. Clear sensory differences were found between beers brewed with Typical harvest Cascade hops and Late harvest Cascade hops, in terms of difference testing, descriptive analysis and consumer preference tests. / Graduation date: 2013

Heat resistant sporeforming psychrotrophic bacteria were isolated from raw milk samples from 59 Grade A farms in Oregon. Forty-nine of the 59 (83%) raw milk samples in this survey contained sporeforming psychrotrophic bacteria; isolates from twenty-four (40%) of the samples exhibited proteolytic properties. Populations of sporeforming psychrotrophic bacteria ranged from <10 to >10,000 CFU/mL for all samples. One hundred-two isolates were identified as Bacillus species. Twelve different Bacillus species were identified with B. licheniformis being the most predominate (18% of the samples) and B. laterosporus the least frequently isolated species, (2%). Fifty-eight percent of the bacilli isolates produced a bitter off-flavor and putrid odor, while 42% produced a fruity and/or rancid off-flavor when inoculated into sterile whole milk. Based on biochemical activity tests, 83% of the thermoduric isolates hydrolysed casein while 56% were proteolytic (in litmus milk), 57% demonstrated lipolytic activity and 31% produced acid in litmus milk. Forty-eight isolates that tested positive for proteolysis were evaluated quantitatively for activity, which ranged from 0.93 to 1.93 units (expressed as mM of alanine). Isolates of Bacillus cereus var. mycoides demonstrated significantly higher (p>0.05) proteolytic activity than other Bacillus species isolated. / Graduation date: 1988

8 leaves of plates in pocket / Bibliography: leaves 148-154 / 154 leaves : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--Dept. of Organic Chemistry, University of Adelaide, 1983

Thesis (MSc)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: ~-Glucosidases constitute a major group of biologically important enzymes that catalyze the hydrolysis of glycosidic linkages in ~-glucosides, as well as in glycosides that contain only carbohydrate residues, e.g. cellobiose. These enzymes occur in all living kingdoms and perform a variety of functions in organisms ranging from bacteria to highly evolved mammals. Three different types of ~-glucosidases are found in humans, each with its own function: glucocerebrosidase (a deficiency causes Gaucher disease), lactase-phlorizin hydrolase (a deficiency results in lactose intolerance) and cytosolic ~-glucosidase (responsible for the hydrolysis of ~- glucosides ingested with foods of plant and animal origin). In plants, the functions of ~-glucosidases include pigment metabolism, biomass conversion and cyanogenesis, a function it shares with insect ~-glucosidases. Microbial ~-glucosidases, as part of the cellulase enzyme system that is responsible for the hydrolysis of cellobiose and short-chain oligosaccharides into glucose, playa role in the conversion of cellulosic biomass to liquid fuel. These microbial ~-glucosidases also playa very important role in the enhancement of fruit and wine aromas through the liberation of monoterpenols. Monoterpenols play an invaluable role in the flavor and aroma of grapes and wine, and are present as free, volatile and odorous molecules, as well as flavorless, non-volatile glycosidic complexes. These complexes most often occur as 6-0-~-Dxvlopyranosyl- B-Dcqlucopyranosides, 6-0-~-D-glucopyranosyl-~-D-glucopyranosides, 6-0-a-L-arabinofuranosyl-~-D-glucopyranosides, 6-0-a-L-rhamnopyranosyl-~-Dglucopyranosides, or 6-0-~-D-apiofuranosyl-~-D-glucopyranosides of mainly linalool, geraniol, nerol, a-terpineol and hotrienol. Two mechanisms exist for the release of monoterpenes from glycosidically bound non-volatile precursors: acid hydrolysis and enzymatic hydrolysis. As high temperature acid hydrolysis causes a rearrangement of the monoterpene aglycones, the focus has shifted towards the more efficient enzymatic hydrolysis that does not result in modifications of the intrinsic aromatic character of the wine. The endogenous ~-glucosidases of grapes (Vitis vinifera), as well as of the wine yeast Saccharomyces cerevisiae, exhibit very low activity towards the glycoside precursors, and thus the focus has increasingly fallen on the addition of exogenous ~-glucosidases to enhance wine flavor. Fungal, bacterial and some yeast ~- glucosidases have been indicated as effective aroma liberators, but these enzymes are not always suitable for use under the harsh conditions that prevail during winemaking (i.e. low pH, low temperatures, and high ethanol and glucose concentrations). The limited enzyme activities of the abovementioned microorganisms have resulted in a search among non-Saccharomyces yeasts for ~- glucosidases that can withstand these conditions. The ~-glucosidase activities of 20 wine-associated non-Saccharomyces yeasts were quantified, characterized and assessed to determine the efficiency with which they could liberate monoterpenols from their terpenyl-glycosides. The Debaryomyces pseudopolymorphus l3-glucosidase from intracellular crude cell extracts exhibited the most suitable combination of properties in terms of functionality at wine pH, resistance to wine-associated inhibitory compounds (glucose, ethanol and sulfur dioxide), high substrate affinity and large aglycone-substrate recognition. This yeast strain was also used, in conjunction with S. cerevisiae VIN13, for the small-scale fermentation of Chardonnay juice. The results indicated that the l3-glucosidase of D. pseudopolymorphus had definite potential as a wine aroma-enhancing enzyme, as the concentrations of free terpenols (nerol, geraniol and citronellol) were significantly increased during fermentation. Future experimental work would include an in-depth study of the kinetic characteristics of the l3-glucosidases (both cytosolic and cell-associated) exhibiting the highest terpenol-liberating activity under winemaking conditions. The next step would then be the cloning and expression of the most efficient l3-glucosidase gene in a commercial wine yeast. Such a recombinant wine yeast would release grapederived aroma compounds from their non-volatile precursors during single culture fermentations, thereby increasing the sensorial quality of wine. / AFRIKAANSE OPSOMMING: I3-Glukosidases vorm deel van 'n groot groep biologies belangrike ensieme wat die hidrolise van glikosidiese bindings binne l3-glukosiede,sowel as binne glikosiede wat slegs uit koolhidraatresidue bestaan, soos bv. sellobiose, kataliseer. Hierdie ensieme kom in alle koningkryke van lewende organismes voor en verrig 'n wye verskeidenheid funksies binne organismes wat wissel van bakterieë tot hoogs ontwikkelde soogdiere. Drie verskillende tipes l3-glukosidases,elk met sy eie funksie, kom in mense voor: glukoserebrosidase ('n gebrek hieraan lei tot Gaucher-siekte), laktaseflorizinhidrolase ('n gebrek hieraan gee aanleiding tot laktose-intoleransie) en sitosol l3-glukosidase (verantwoordelik vir die hidrolise van l3-glukosiede wat saam met voedsel van plant en dier oorsprong ingeneem word). Die funksies van 13- glukosidase binne plante sluit in pigmentmetabolisme, biomassa-omsetting en sianogenese, wat ook 'n funksie van insek l3-glukosidases is. Mikrobiese 13- glukosidases, as deel van die sellulase-ensiemsisteem wat verantwoordelik vir die hidrolise van sellobiose en kortketting-oligosakkariede na glukose is, speel 'n rol in die omsetting van sellulosebiomassa na brandstof. Hierdie mikrobiese 13- glukosidases speelook 'n baie belangrike rol in die verbetering van vrugte- en wynaroma deur die vrystelling van monoterpenole. Monoterpenole speel 'n belangrike rol in die geur en aroma van druiwe en wyn, en kom voor as vry, vlugtige en aromatiese molekules, asook geurlose, nie-vlugtige glikosidies-gebonde komplekse. Hierdie komplekse is meestal in die vorm van 6-0- I3-D-xilopiranosiel-I3-D-glukopiranosiede, 6-0-a-L-arabinofuranosiel-I3-D-glukopiranosiede, 6-0-I3-D-glukopiranosiel-I3-D-glukopiranosiede, 6-0-a-L-ramnopiranosiel- I3-D-glukopiranosiede,of 6-0-I3-D-apiofuranosiel-I3-D-glukopiranosiedevan hoofsaaklik linalool, geraniol, nerol, a-terpineol en hotrienol. Monoterpenole kan op een van twee maniere van hul suikermolekules vrygestel word: suurhidrolise of ensimatiese hidrolise. Die hoë temperature waarby suurhidrolise plaasvind veroorsaak 'n herrangskikking van die monoterpeen aglikone, en die fokus het gevolglik verskuif na die meer effektiewe ensimatiese hidrolise wat nie verandering van die intrinsieke aromatiese karakter van die wyn tot gevolg het nie. Die endogene l3-glukosidases van druiwe (Vitis vinifera) en die wyngis Saccharomyces cere visiae , toon baie lae aktiwiteit ten opsigte van die aromatiese voorlopers, en dus word daar toenemend op die toevoeging van eksogene 13- glukosidases tot die wyn gefokus om meer geur vry te stel. Daar is bevind dat 13- glukosidases van fungiese, bakteriële en gis oorsprong effektiewe aromavrystelIers is, maar hierdie ensieme is nie altyd gepas vir gebruik in wyn nie, aangesien dit 'n omgewing is met 'n lae pH, lae temperatuur, en hoë etanol- en glukosekonsentrasies. Die beperkte ensiemaktiwitiet van bogenoemde mikroorganismes het gelei tot 'n soeke onder nie-Saccharomyces giste na l3-glukosidases wat in die wynomgewing kan funksioneer. Die ~-glukosidase-aktiwiteit van twintig wyn geassosieerde nie-Saccharomyces giste is gekwantifiseer en gekarakteriseer om te bepaal tot watter mate dit monoterpenole van hul terpeniel glikosiede kan vrystel. Die intrasellulêre ~- glukosidase teenwoordig in the selekstrak van Debaryomyces pseudopolymorphus, het die belowendste resultate getoon ten opsigte van funksionaliteit by wyn se pH, weerstand teen wyn geassosieerde inhibeerders (glukose, etanol en swaweidioksied), hoë substraataffiniteit en breë aglikoon-substraat herkenning. Hierdie gisras is ook in kombinasie met S. cerevisiae VIN13 gebruik vir die kleinskaalse fermentasie van Chardonnay sap. Die resultate het getoon dat die ~- glukosidase van D. pseudopolymorphus wel potensiaal het om wynaroma te verhoog, aangesien die konsentrasie van ongebonde terpenole (nerol, geraniol en citronellol) aansienlik tydens fermentasie toegeneem het. Toekomstige eksperimentele werk sluit in, onder meer, In in-diepte studie van die kinetiese eienskappe van die ~-glukosidases (beide sitesolies en sel-geassosieerd) wat die meeste terpenole onder wynrnaakkondisies vry stel, asook die klonering en uitdrukking van die enkele ~-glukosidasegeen met die hoogste aktiwiteit, in In kommersiële wyngis. Só In rekombinante wyngis sal die vrystelling van druifgebaseerde aromakomponente van hul nie-vlugtige, geurlose voorlopers tydens enkel-kultuur fermentasies teweeg bring.

This study was conducted to determine the effect of heating methods (conventional and microwave), heating temperatures, and refrigerated storage on the flavor deterioration in precooked chicken parts. Chicken breasts and legs were evaluated by chemical analyses and sensory evaluation. Data was analyzed by Fisher's least significant difference (LSD) and Duncan's test. Results of the thiobarbituric acid test (TBA) indicated that heating temperature, regardless of heating method had a significant influence on oxidative deterioration. When the legs were heated for a longer time at a lower temperature, the TBA values significantly increased indicating heightened warmed-over flavor (WOF). Two-day refrigerated storage had no measurable influence on the TBA numbers. The legs generally had a higher degree of lipid oxidation reflected by higher TBA values. Heating method and 2-day refrigerated storage did not have a significant effect on the nonheme iron content of breasts or legs. The effects of heating temperature were inconsistent for the legs and breasts which was attributed to the different initial iron content of the legs, and the release of nonheme iron during the initial processing of the legs. The nonheme iron values of the legs were greater relative to the breasts. Heating and storage in general increased the area of peak 3 in legs, but had no effect on the breasts. A significant increase in the hexanal (peak 8) content of legs occurred upon 2-day storage, but not for breasts. Peak 7 significantly decreased when the breasts were heated, stored, and reheated. This was not the case for the legs. Sensory panelists could not differentiate between heating methods or temperatures for either part. / Master of Science

Orientador: Maria Aparecida A. P. da Silva / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-05T15:54:43Z (GMT). No. of bitstreams: 1 Almeida_SelmaBergara_D.pdf: 1651470 bytes, checksum: a5ee60dbc52c3dc1aeab330822f4380b (MD5) Previous issue date: 2006 / Resumo: Para a recuperação do aroma e sabor de fruta fresca ao suco industrializado, subprodutos do processamento do suco de laranja, como óleo essencial e essências aquosa e oleosa, são normalmente reincorporados ao produto processado. A adição de cada um desses subprodutos confere ao produto final qualidade e estabilidade sensoriais distintas. O presente trabalho teve por objetivo avaliar a qualidade e estabilidade sensoriais do suco de laranja reconstituído, produzido com laranjas de variedades brasileiras e aromatizado com quatro diferentes aromas naturais de laranja, quais sejam: I - óleo essencial; II - frações destiladas de essência oleosa (oil phase); III - aroma formulado contendo frações destiladas de óleo essencial e de essência oleosa; e, IV - aroma formulado contendo frações destiladas de óleo essencial, frações destiladas de essência oleosa e de essência aquosa (water phase). Assim, esta pesquisa consistiu em quatro etapas, a saber: avaliação do impacto dos quatro aromas naturais sobre a aceitação e perfil sensorial do suco de laranja pronto para beber em condições pré-processamento térmico; avaliação da estabilidade sensorial dos aromas naturais ao processamento térmico do suco; avaliação do impacto dos aromas naturais sobre a estabilidade sensorial do suco pronto para beber durante o armazenamento da bebida e, finalmente, a identificação de compostos voláteis odoríferos presentes em um dos aromas mais estáveis - Aroma III. Quatro formulações de suco de laranja reconstituído foram elaboradas, adicionando-se os aromas I, II, III e IV, processadas e envasadas assepticamente em garrafas PET. Os perfis sensoriais, através de Análise Descritiva Quantitativa, e a aceitação das quatro formulações de suco de laranja pronto para beber foram avaliados antes e após o processamento térmico, bem como durante o armazenamento da bebida por 20, 40, 60 e 90 dias. Índices de escurecimento e teores de vitamina C foram determinados durante o armazenamento. Adicionalmente, amostras comerciais de suco de laranja pronto para beber foram avaliadas. Os dados foram analisados por ANOVA, teste de média Tukey ou t-Student, Mapa de Preferência, Análise de Componentes Principais, análise de correlação de Pearson e análise de regressão linear dos parâmetros sensoriais em função do tempo de armazenamento da bebida. O aroma III foi avaliado por cromatografia gasosa (CG), CG-olfatometria (Osme) e espectrometria de massas. Os resultados obtidos na primeira etapa da pesquisa sugeriram que a reincorporação dos compostos voláteis provenientes da essência oleosa contribuem menos para aumentar a aceitação do suco de laranja do que aqueles provenientes do óleo essencial. Em contrapartida, as amostras comerciais tiveram aceitação modesta entre os consumidores indicando que, no Brasil, os sucos de laranja comerciais podem ainda ter sua qualidade sensorial melhorada pela adição de novos aromas naturais. Na segunda etapa, a avaliação da aceitação das amostras pré e pós-processamento indicou boa estabilidade dos aromas naturais estudados ao processamento térmico. Os perfis sensoriais das formulações sugeriram que: i) a reincorporação ao suco de frações destiladas dos três subprodutos do processamento da laranja, resulta em maior estabilidade térmica do que aromas derivados apenas do óleo essencial; e, ii) a presença de voláteis provenientes da essência aquosa no aroma natural de laranja, mesmo que em proporções minoritárias, realça no suco notas de sabor de suco natural e suaviza as notas de aroma e sabor de casca de laranja. Na terceira etapa, durante o armazenamento da bebida, a formulação contendo apenas frações de essência oleosa apresentou, notadamente, menor estabilidade sensorial do que as demais formulações, contendo compostos voláteis provenientes do óleo essencial. Possivelmente essa ocorrência seja devida à presença de agentes antioxidantes naturais nas frações derivadas do óleo essencial. Adicionalmente, os resultados sugeriram que a adição ao suco de laranja de frações destiladas de óleo essencial resulta em melhor estabilidade sensorial à bebida do que a adição ao suco do óleo essencial na sua forma integral. Em todas as amostras, ao longo do período de armazenamento, ocorreu queda dos teores de vitamina C e concomitante escurecimento dos sucos. Finalmente, na última etapa, foram gerados 55 picos de odores, os quais apresentaram, predominantemente, notas de aroma associadas à laranja. Beta-mirceno, butanoato de etila, a-pineno, hexanal, citral e decanol foram os compostos voláteis de maior importância odorífera identificados. Os compostos ß-terpineol, citronelal, undecanal, acetato de a-terpenila, acetato de nerila e a-cariofileno foram identificados como compostos de alto poder odorífero e, limoneno, linalol, a-terpineol e decanal, como compostos de baixo poder odorífero / Abstract: To restore the natural orange aroma and flavor to industrialized juices, processors add natural flavorings obtained from three orange juice by-products: the cold-pressed oil, the oil phase and the water phase. The addition of each of these by-products results in juices with distinct sensory characteristics and acceptability. The present work aimed at evaluating the sensory quality and stability of reconstituted orange juice from Brazilian varieties, flavored with four different orange natural aromas, which are: I ¿ essential oil; II ¿ distilled fractions of the oil phase; III - formulated aroma containing distilled fractions of the essential oil and of the oil phase e, IV ¿ formulated aroma containing distilled fractions of the essential oil, distilled fractions of the oil phase and of the water phase. Thus the present research was divided into four stages: evaluation of the impact of four natural aromas (I, II, III, IV) on the acceptability and sensory profile of ready-todrink orange juice before thermal treatment; evaluation of the sensory stability of the natural aromas to the thermal treatment of the juice; evaluation of the natural aromas impact to the sensory stability of ready-to-drink orange juice during storage; the identification of odor volatiles compounds in one of the most stable aromas ¿ Aroma III. Four reconstituted orange juices formulations were elaborated, by adding the above mentioned flavorings (I, II, III, IV), processed and packed in PET bottles. The sensory profiles, by using Quantitative Descriptive Analysis, and the acceptability of the four ready-to-drink orange juices were evaluated before and after the processing, and also during their 20, 40, 60 and 90 day-storage. In addition, other commercial samples of ready-to-drink orange juice were included in this study. Determinations of the ascorbic acid and the browning index of formulated juices were done as well. Data were analyzed by ANOVA, Tukey or t-Student mean test, Principal Component Analysis, Preference Mapping, Pearson¿s correlation analysis and linear regression analysis of sensory parameters as a function of the storage time. The aroma III was evaluated by gas chromatography (GC), GC-olfactometry (OSME) and by mass spectrometry. The results obtained from the first research stage suggested that reintroduction of the volatiles compounds from oil phase contribute less to increase the acceptability of orange juice than those from the essential oil. On the other hand, all the commercial samples presented moderate acceptability by consumers, indicating that the commercial orange juices in Brazil may have their sensory quality improved by adding new natural aromas. In the second research stage, the evaluation of the pre and post-processing samples acceptability indicated that the studied natural aromas presented good stability to the thermal treatment. The formulations sensory profiles suggested that: i) the reintroduction of the distilled fractions of the three orange processing by-products to the juice results in higher thermal stability than that caused by the aromas derived from the essential oil; ii) the presence of volatiles from distilled fractions of the water phase in the natural orange aroma, although in very small proportions, enhances natural juice flavor and softens orange peel flavor and aroma. In the third research stage, during the drink storage the formulation containing only distilled fractions of the oil phase presented much less sensory stability than the other formulations, containing volatile compounds from the essential oil. A possible explanation for this occurrence is the presence of natural antioxidants in those fractions derived from essential oil. Further, the results suggested that the addition of distilled fractions of essential oil to orange juice provides better sensory stability to the drink than the addition of the integral essential oil. During storage all samples showed decrease in vitamin C and simultaneous browning of juices. Finally, in the last research stage, Fifty-five odor peaks were generated, which presented aroma notes associated mostly to orange. Beta-myrcene, ethyl butanoate, a-pinene, hexanal, citral and decanol were identified as the highest aroma impact compounds. The compounds ß-terpineol, citronellal, undecanal, a-terpinyl acetate, neryl acetate and a-cariophylene were identified as high odor power compounds and, limonene, linalool, a-terpineol and decanal, as low odor power compounds / Doutorado / Doutor em Tecnologia de Alimentos

Thesis (MSc)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT: Monoterpene alcohols (monoterpenols) play an important role in the flavour and aroma of grapes and wine. This is especially applicable to wines of a muscat variety, but these flavour compounds are also present in other non-muscat grape varieties, where they supplement other varietal flavours and aromas. These monoterpenols can be found in grapes and wine as free, volatile and odorous molecules, as well as in flavourless, nonvolatile glycosidic complexes. These complexes most often occur as 6-0-a-L-arabinofuranosyl-p-D-glucopyranosides (vicianosides), 6-0-P-D-xylopyranosyl- P-D-gluco-pyranosides (primverosides), 6-0-P-D-glucopyranosyl-p-D-glucopyranosides (gentio-biosides ), 6-0-a-L -rhamnopyra nosyl-p-D-g lucopyra nos ides (rutinos ides), or 6-0-p-D-apiofuranosyl-p-D-glucopyranosides of mainly linalool, geraniol, nerol, a-terpineol and hotrienol. These precursors are, however, hydrolyzed only to a limited extent by endogenous glycosidases during the fermentation process, as they exhibit very low activity in wine conditions. The monoterpenols can be released from their sugar moieties by one of two methods: either an acid or an enzymatic hydrolysis. The enzymatic hydrolysis mechanism is fully understood, and the process functions in two successive steps: firstly, depending on the precursor, the glycosidic linkage is cleaved by an a-L-arabinofuranosidase, an a-L-rhamnosidase, a p-D-xylosidase, or a p-D-apiosidase. The second step involves the liberation of the monoterpene alcohol by a p-glucosidase. This enzymatic hydrolysis does not influence the intrinsic aromatic characteristics of the wine, as opposed to acid hydrolysis. As the endogenous grape glycosides of Vitis vinifera and the yeast Saccharomyces cerevisiae show very low activity towards these aromatic precursors during the handling of the juice and winemaking processes, the focus has increasingly fallen on introducing exogenous p-glucosidases to wines and juices. Genes encoding p-glucosidases and a-L-arabinofuranosidases have been cloned from various organisms, including bacteria, fungi and yeasts. However, the activities and properties of these enzymes are not always suitable for exploitation under winemaking conditions, where a low pH, low temperatures, and high ethanol and glucose concentrations prevail. A genetically engineered wine yeast strain of S. cerevisiae that expresses glycosidases that are active in these conditions would be useful in improving the flavour and aroma of wines, thereby adding to the complexity and value of the wine. Two p-glucosidase genes, BGL 1 and BGL2 from Saccharomycopsis fibufigera, were subcloned into two Escherichia coli-yeast shuttle vectors. A dominant selectable marker gene (SMR1) was also inserted onto these plasmids. These plasmids were designated pBGL 1 (containing the BGL 1 gene) and pBGL2 (containing the BGL2 gene) respectively. Introduction of the two plasmids into two strains of S. cerevisiae then followed. A laboratory strain, L1278, was transformed to confirm the effective secretion of the expressed protein. An industrial yeast strain, VIN13, was subsequently transformed by making use of the selectable marker (resistance against sulfometuron). Enzyme assays with the synthetic substrate p-nitrophenol-j3-D-glucopyranoside (pNPG) were performed to determine the activity of the j3-glucosidases over a period of days, as well as at certain temperatures and pH values. The stability of the enzymes was also investigated. These recombinant yeasts were able to degrade the pNPG efficiently. They showed promising results concerning pH optima, with a substantial amount of activity found at the pH levels as found in the wine environment. There was also a slight increase in specific activity at lower temperatures. The recombinant yeast strains were also tested in smallscale fermentations. Three wines were made, of which two were from white cultivars (Chenin blanc and GewOrtztraminer) and one from red (Pinotage). Results obtained from micro-extraction from the finished wines showed that the terpenol content did increase, although this was not the only wine component influenced. Other flavour compounds also showed increases, especially the esters. This also played a role in the flavour increase in the wine. Future work would include optimizing the available results. This would entail the addition of another glycosidic enzyme, such as a-L-arabinofuranosidase, to the genome of the wine yeast to aid the further breakdown of glycosidic bonds. The cloning or engineering of a j3-glucosidase enzyme that is more active at low temperatures would also yield better results and release even more of the aroma of the wine. / AFRIKAANSE OPSOMMING: Monoterpeenalkohole (monoterpenole) speel 'n belangrike rol in die geur en aroma van druiwe en wyn. Dit is veral van toepassing op wyne van Muskaat-varieteite, maar hierdie geurkomponente is ook teenwoordig in ander nie-Muskaat druifsoorte, waar dit bydra tot die varieteitsqeur en aroma. Hierdie monoterpenole kom voor in druiwe as vry, vlugtige en aromatiese molekules, of as geurlose, nie-vlugtige glikosidies-gebonde komplekse. Hierdie komplekse is meestal in die vorm van 6-0-a-L-arabinofuranosiel-~-D-glukopiranosiede, 6- O-~-D-xilopiranosiel-~-D-glukopiranosiede (primverosiede), 6-0-~-D-glukopiranosiel-~-Dglukopiranosiede (gentiobiosiede), 6-0-a-L-ramno-pyranosiel-~-D-glukopiranosiede (rutinosiede), of 6-0-~-D-apiofuranosiel-~-D-glukopirano-siede van hoofsaaklik linalool, geraniol, nerol, a-terpineol en hotrienol. Hierdie geurvoorlopers word egter slegs tot In beperkte mate tydens die proses van fermentasie deur die endogene glikosidase ensieme gehidroliseer, aangesien hulle baie min aktiwiteit toon onder wynbereidingstoestande. Die monoterpenole kan op een van twee wyses van hul suikermolekules vrygestel word: 'n suurhidrolise, of ensiematiese hidrolise. Die ensiematiese hidroliseproses word baie goed begryp en behels twee opeenvolgende stappe: eerstens, afhangende van die aard van die voorloper, word die glikosidiese verbinding deur In a-L-arabinofuranosidase, In a-L-ramnosidase, In ~-D-xilosidase, of 'n ~-D-apiosidase gebreek. In die tweede stap word die monoterpeenalkohol deur In ~-glukosidase vrygestel. Hierdie ensiematiese afbraakproses verander nie die intrinsieke aromatiese kenmerke van die wyn, soos wat met suurhidrolise die geval is nie. Omdat die endogene glikosidases van Vitis vinifera en die van die gis Saccharomyces cerevisiae baie lae aktiwiteit ten opsigte van die aromatiese voorlopers gedurende die hantering van die druiwesap en wynmaakprosesse toon, val die fokus al hoe meer op die inkorporering van eksogene ~-glukosidases in wyn en sappe. Gene wat vir ~- glukosidases en a-L-arabinofuranosidases kodeer, is al vanuit verskeie organismes gekloneer, insluitende bakteriee, fungi en giste. Die aktiwiteite en kenmerke van hierdie ensieme is egter nie altyd wenslik vir hul gebruik in wyn nie, aangesien dit In omgewing is met 'n lae pH, lae temperatuur, en hoe etanolvlakke en glukose-konsentrasies. In geneties veranderde wyngis van S. cerevisiae wat in staat is om glikosidases uit te druk wat onder hierdie kondisies aktief is, sal baie handig te pas kom in die verbetering van die geur en aroma van wyne, om daardeur die kompleksiteit en die waarde van die wyn te verhoog. Twee ~-glukosidasegene, BGL 1 en BGL2 vanaf die gis Saccharomycopsis fibuligera , is in twee afsonderlike Esccherichia coli-gis-pendelplasmiede gesubkloneer. In Dominante selekteerbare merkergeen (SMR1) is ook in hierdie plasmiede gekloneer. Hierdie plasmiede word onderskeidelik pBGL 1 (met die BGL 1-geen) en pBGL2 (bevattende die BGL2-geen) genoem. Hierdie twee plasmiede is hierna apart na twee rasse van S. cerevisiae getransformeer. Eerstens is 'n laboratoriumras, L1278, getransformeer om te bevestig dat effektiewe sekresie en uitdrukking van die proteTen plaasvind. Hierna is 'n industriele gisras, VIN13, getransformeer deur gebruik te maak van die selektiewe merker (bestandheid teen sulfometuron). Ensiem-bepalings met behulp van die sintetiese substraat p-nitrofeniel-p-O-glukopiranosied (pNPG) is gedoen om die aktiwiteit van die p-glukosidqses oor 'n aantal dae te bepaal, asook om die aktiwiteit by sekere temperature en pH-vlakke te meet. Die stabiliteit van die ensieme is ook bepaal. Hierdie rekombinante giste was in staat om pNPG effektief af te breek. Hulle het belowende resultate betreffende die pH-optima getoon, met 'n aansienlike hoeveelheid aktiwiteit by die pH-vlakke soos dit in die wynomgewing voorkom. Daar was ook 'n effense verhoging in die ensieme se aktiwiteite by laer temperature. Die rekombinante gisrasse is ook in kleinskaalse wynfermentasies gebruik. Drie verskillende wyne is gemaak, waarvan twee wit kultivars was (Chenin blanc en GewOrtztraminer) en een 'n rooi kultivar (Pinotage). Resultate wat deur die mikro-ekstraksie van die voltooide wyne verkry is, het getoon dat die terpenolinhoud wei verhoog het, alhoewel dit nie die enigste wynkomponente was wat beinvloed is nie. Ander geurkomponente het ook 'n verhoging in konsentrasie getoon, veral die esters. Hierdie verbindings het ook 'n rol in die verhoging van geur in die wyne gespeel. Toekomstige werk sal die beskikbare resultate verder optimaliseer. Dit sal insluit die byvoeging van nog 'n glikosidiese ensiem, soos a.-L-arabinofuranosidase, tot die genoom van die wyngis, om verdere afbraak van glikosidiese verbindings teweeg te bring. Die klonering of verandering van 'n p-glukosidase-ensiem met verhoogde aktiwiteit by laer temperature sal ook beter resultate toon en meer geur in die wyn kan vrystel.

Thesis (PhD)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: Yeasts produce a broad range of aroma-active volatile esters and higher alcohols during alcoholic fermentation. Some of these esters and higher alcohols are important for the fruity flavors and therefore the final quality of wine and other fermented beverages. Esters are produced and hydrolyzed by alcohol acetyltransferases and esterases, respectively. In yeast, ester-synthesizing activities are represented by two alcohol acetyltransferases encoded by the ATFI and ATF2 genes, and by an ethanol hexanoyl transferase encoded by the EHTI gene. Atfl p and Atf2p appear responsible for the production of ethyl acetate and isoamyl acetate, while Ehtl p synthesizes ethyl hexanoate from ethanol and hexanoyl-CoA. Although a fair amount of information is available regarding the ATF 1 gene, limited information is available on the remaining alcohol acetyltransferases. Only two genes that code for esterases have been identified in yeast, namely lAHI and TIPI. It has also been shown that the balance between alcohol acetyltransferases and esterases is important for the net rate of ester accumulation. Higher alcohols are synthesized from the a-keto-acids in the branched-chain amino acid metabolic pathway by decarboxylation and reduction. The transamination of the amino acid to the respective a-keto-acid is catalyzed by mitochondrial and cytosolic branched-chain amino acid transferases, which are encoded by the BATI and BAT2 genes, respectively. In recent years, a strong scientific and industrial interest in the metabolism of flavoractive compounds has emerged, but information regarding the roles of specific enzymes and the physiological relevance of their metabolism remains limited. The aim of this project was to investigate the physiological and metabolic consequences of changes in the expression levels of some of the key enzymes involved in aroma compound production. The consequences of these changes on the chemical composition and the fermentation bouquet of wines and distillates were also investigated. The first part of the section on the results in this dissertation reports on the role and relative importance of the Saccharomyces cerevisiae enzymes involved in ester metabolism, namely Atflp, Atf2p, Ehtlp, Iahlp and Tiplp. The corresponding genes were overexpressed in a laboratory strain of S. cerevisiae, BY4742, and in a widely used commercial wine yeast strain, VIN13. Table wine and base wines for distillation were prepared with these VIN13 transformed strains. The ester concentrations and aroma profiles of the wines and distillates were analyzed and compared. The data indicated that the overexpression of ATF 1 and ATF2 increased the concentrations of ethyl acetate, isoamyl acetate, 2-pheylethyl acetate and ethyl caproate, while the overexpression of JAHI resulted in a significant decrease in the concentrations of ethyl acetate, isoamyl acetate, hexyl acetate and 2-phenylethyl acetate. The overexpression of EHTI resulted in a marked increase in the concentrations of ethyl caproate, ethyl caprylate and ethyl caprate, while the overexpression of TJP1 did not decrease the concentrations of any of the esters. In most cases, there was a correlation between the increase in esters and the decrease in higher alcohols. The data suggest that yeast balances the amount of different esters produced through alcohol acetyltransferases and esterases, and that, in some cases, these enzymes appear to overlap in function and/or influence each other's activity. In the second part of the results section, the consequences of the deletion and the overexpression of two genes, BATl and BAT2, which encode transaminases that contribute to the metabolism of higher alcohols, were investigated. The genes were both disrupted in a S. cerevisiae BY4742, and overexpressed in both this laboratory strain and in the VIN13 wine yeast strain. The effects of these modifications on the general physiology of the corresponding yeast strains and on higher alcohol metabolism were assessed in a range of growth conditions, including aerobic and anaerobic growth conditions, in the presence of glucose or raffinose as sole carbon source and growth in the presence of various concentrations of amino acids. Table wine and base wines for distillation were prepared with the modified industrial strains and the concentrations of the higher alcohols and the aroma profiles of the wine and distillates were analyzed and compared. Batl deletion seemed to be lethal under the conditions that were created, and therefore only the bat2!:!.strain, together with the BATI and BAT2 overexpression strains, were investigated. These modifications did not appear to significantly affect the general physiology of the strains. The results obtained indicated that the overexpression of BATI increased the concentrations of isoamyl alcohol and isoamyl acetate, and, to a lesser extent, the concentrations of isobutanol and isobutyric acid. The overexpression of the BAT2 gene resulted in a substantial increase in the levels of isobutanol, isobutyric acid and propionic acid production, and a modest increase in the level of propanol and isovaleric acid. Interestingly, the overexpression of BAT2 led to a decrease in isoamyl alcohol and isoamyl acetate concentrations. Sensory analyses indicated that the wines and distillates produced with the strains in which the BATl and BAT2 genes were overexpressed had more fruity characteristics (peach and apricot aromas) than the wines produced by the wild-type strains. This study offers new prospects for the development of wine yeast starter strains with optimized ester and higher alcohol-producing capability that could assist winemakers in their efforts to consistently produce wine to definable specifications and styles and a predetermined flavor profile. / AFRIKAANSE OPSOMMING: Gedurende fermentasie produseer giste 'n wye verskeidenheid vlugtige aromatiese esters en hoër alkohole. Sommige van hierdie esters en hoër alkohole is belangrik vir die vrugtige geure en dra dus by tot die finale kwaliteit van wyn en ander gefermenteerde drankies. Esters word onderskeidelik deur alkoholasetieltranferases en esterases geproduseer en gehidroliseer. In giste word die ester-sintetiserende aktiwiteite deur twee alkoholasetieltransferases verteenwoordig wat deur die ATFI-en ATF2-gene, asook 'n etanolheksanoïeltransferase wat deur die EHTl-geen, gekodeer word. Dit blyk dat ATFlp en ATF2p verantwoordelik is vir die produksie van etielasetaat en isoamielasetaat, terwyl Ehtl p-etielheksanoaat vanaf etanol en heksanoïel-CoA sintetiseer. Alhoewel daar 'n redelike hoeveelheid inligting t.o.v die ATF I-geen beskikbaar is, is daar weinig inligting oor die res van die aloholasetieltransferases. Slegs twee gene wat vir esterases kodeer, is in gis geïdentifiseer, naamlik IAHI en TIPI. Daar is ook bewys dat 'n balans tussen die alkoholasetieltransferases en esterases baie belangrik is vir die netto-tempo van ester-akkumulasie. Hoër alkohole word gesintetiseer vanaf a-keto sure in die vertakte-ketting aminosuur metaboliese pad deur dekarboksilasie en reduksie. Die transaminasie van die aminosuur na die onderkeidelike a-ketosuur word deur vertakte-ketting aminosuur transferases, geleë in die mitochondrion en sitosol, en gekodeer deur BATl- en BAT2-gene, gekataliseer. In die laaste paar jare het daar 'n sterk wetenskaplike, asook industrïele, belangstelling in die metabolisme van aroma-aktiewe komponente te voorskyn gekom, maar inligting in verband met die rol van spesifieke ensieme en die fisiologiese belangrikheid van hul metabolisme is egter beperk. Die doel van hierdie projek was om die fisiologiese en metaboliese gevolge van veranderinge in die ekspressievlakke van sommige sleutelensieme betrokke by aromakomponent-produksie te ondersoek. Die gevolge van hierdie veranderinge op chemiese vlakke, asook hoe die fermentasie-aroma van die wyne en distillate beïnvloed word, is ook bestudeer. Die eerste gedeelte van die resultate rapporteer oor die rol en relatiewe belangrikheid van die Saccharomyces cerevisiae-ensieme betrokke by estermetabolisme, naamlik Atfl p, Atf2p, Ehtlp, Iahlp en Tiplp. Die gene was ooruitgedruk in 'n laboratoriurnras van S. cerevisiae, BY4742, asook in 'n kommersïele wyngisras, VIN13. Tafelwyne en basiswyne vir distillasie is gemaak met die getransformeerde VIN13-rasse. Die esterkonsentrasies en aromaprofiele van die wyne en distillate is ontleed en vergelyk. Die data het gewys dat die ooruitdrukking van ATFI- en ATF2-gene 'n verhoging in etielasetaat, isoamielasetaat, 2-fenieletielasetaat en etielkaproaat veroorsaak het, terwyl ooruitdrukking van !AHI 'n betekenisvolle afname in etielasetaat-, isoamielasetaat-, heksielasetaat- en 2-fenieletielasetaat-konsentrasies veroorsaak het. Die ooruitdrukking van EHTI het 'n duidelike verhoging in etielkaproaat, etielkaprilaat en etielkapraat veroorsaak en die ooruitdrukking van TIPIhet geen van die esterkonsentrasies verander nie. In die meeste gevalle was daar nie 'n korrelasie tussen die toename in esters en afname in hoër alkohole nie. Die data stelook voor dat die gis 'n balans tussen die verskillende esters handhaaf deur middel van die alkoholasetieltrasferases en esterases, en in sommige gevalle blyk dit dat die ensieme dieselfde funksies het en/of mekaar se aktiwiteit beïnvloed. In die tweede gedeelte van die resultate is die oorsake van delesie en ooruitdrukking van twee gene, BAT1 en BAT2, wat kodeer vir transaminases wat tot hoër alkohol metabolisme bydra, bestudeer. Die gene is uitgeslaan in S. cerevisiae BY4742 en ooruitgedruk in BY4742 en in die wyngisras VIN13. Die effekte van hierdie modifikasies op die algemene fisiologie van die verskillende gisrasse en op hoëralkoholmetabolisme is onder 'n verskeidenheid kondisies bestudeer, naamlik aërobies en anaërobiese groeikondisies, in die teenwoordigheid van glukose of raffinose as die enigste koolstofbron, asook in die teenwoordigheid van 'n verskeidenheid konsentrasies aminosure. Tafelwyne en basiswyne vir distillasie is gemaak met die gemodifiseerde industrïele rasse en die konsentrasies van die hoër alkohole en aromaprofiele van die wyne en distillate is ontleed en vergelyk. Bat1-delesie was dodelik onder die kondisies, daarom is slegs die batlts-tes tesame met die BAT1 en BAT2 wat in die rasse ooruitgedruk is, bestudeer. Die modifikasies het nie 'n beduidende effek op die algemene fisiologie van die rasse getoon nie. Die data het wel getoon dat die ooruitdrukking van BAT1 'n verhoging in isoamielalkohol- en isoamielasetaatkonsentrasies, en tot 'n mindere mate isobutielalkohol- en isobottersuur-konsentrasies, veroorsaak het. Die ooruitdrukking van BAT2 het 'n beduidende toename in isobutanol-, isobottersuur- en propioonsuurkonsentrasies en 'n kleinere toename in propanol- en isovaleriaansuur veroorsaak. Die ooruitdrukking van BAT2 het ook gelei tot 'n afname in isoamielalkohol- en isoamielasetaatkonsentrasies. Sensoriese analises het getoon dat die wyne en distillate wat geproduseer is met die rasse waarin die BAT1 en BAT2 gene ooruitgedruk is meer vrugtige eienskappe (perske- en appelkoos-aromas) getoon het as die wyne wat deur die wildetipe rasse geproduseer is. Die studie lewer nuwe vooruitsigte vir die ontwikkeling van wyngiste met geoptimiseerde ester en hoër alkohol produserende eienskappe wat die wynmakers in staat kan stelom wyne te produseer met gedefinieerde spesifikasies en style en 'n voorafbepaalde aromaprofiel.