Doctor of Philosophy / Department of Animal Sciences and Industry / John A. Unruh / Two experiments were conducted to evaluate the effects of exercise on feedlot cattle well-being, growth performance, and carcass characteristics (Experiments 1 and 2). Additionally, two experiments assessed the volatile compound profiles of beef in response to various postmortem processes (Experiments 3 and 4). Experiment 1 compared heifers (n=30) assigned to either a sedentary treatment or an exercise regimen 3 times/wk (20 min/d for the first 2 wk, 30 min/d for the next 2 wk, and 40 min/d for the final wk). Exercised heifers showed decreased blood insulin, daily gains, final body weights, and lower numerical yield grades in conjunction with decreased a* and b* color values of longissimus muscle lean. Experiment 2 exposed steers (n=419) to 1 of 4 treatments; not exercised (CON), or exercised 3 times/wk by animal handlers for 20-30 min for the first 10 wk (EARLY), the last 7 wk (LATE), or for the entire 116 d finishing period (ALL). EARLY treatment cattle exhibited a decrease in blood insulin while EARLY and ALL treatment cattle produced carcasses with decreased marbling scores in comparison to CON cattle. Experiment 3 assessed the volatile compounds generated by raw and cooked beef gluteus medius steaks (n=30) subjected to blade tenderization and aging times ranging from 5 to 61 d. Raw steaks aged longer than 19 d produced greater concentrations of heptanal, octanal, and nonanal than raw steaks aged 5 d, but cooked steaks showed no change in these compounds in response to aging. Additionally, blade tenderization reduced the concentrations of lipid oxidation and Maillard reaction products in cooked steaks. Experiment 4 investigated cross-sections, external, and internal locations of beef longissimus lumborum steaks (n=54) cooked to 63, 71, or 77 °C and aged for 5, 21, or 37 d. External locations generated aldehydes in greater amounts than internal locations while pyrazines are produced exclusively at external locations. Increases in degree of doneness increased all aldehydes, except nonanal, in steak cross-sections. Aldehydes, except hexanal and octanal, generated from external locations became similar to internal locations as aging times increased. Pyrazines from external locations were reduced by increased aging times.

Doctor of Philosophy / Department of Human Nutrition / Delores H. Chambers / Vegetables are popular among consumers because of their versatility of preparation, unique sensory characteristics, and exceptional health benefits. Trends such as organic farming and breeding to increase nutrition and functional health components have increased interest in understanding the flavor of vegetables, such as leafy greens. A lexicon of thirty-two flavor attributes was created to help describe the flavor of fresh leafy vegetables. This lexicon includes five “green” attributes; mouth feel characteristics such as pungent, bite, tooth-etch, and heat/burn; fundamental tastes including bitter and umami; seven terms that describe unique flavors related to specific vegetables such as cabbage, celery, lettuce, spinach, parsley, beet, and radish leaves; and a group of other terms including citrus, piney, woody, water-like, musty/earthy, floral, sulfur, metallic, soapy, petroleum-like, and overall sweet. In addition, our study encompassed a series of sensory tests which will aid in better understanding the effects of several production variables on the sensory characteristics of pac choi and tomato. Variables evaluated were production systems (i.e. organic and conventional), fertilizer amount (i.e. high, low, and no fertilizer), environment (i.e. field and high tunnel), maturity level (i.e. 2.5, 4.5, and 6.5-week old plants at the time of harvest), and shelf life (i.e. 1, 4, 9, 18 days of refrigerated storage). Samples were grown at the Kansas State University Horticulture Research Center located in Olathe, Kansas. Highly trained descriptive panelists from the Sensory Analysis Center at Kansas State University evaluated the samples. There do not appear to be major sensory differences between organic and conventional products specific to the crops and seasons studied. Furthermore, when differences were present, they generally were quite small and showed no clear trends or patterns favoring one production system over the other even after refrigerated storage. However, it is suggested that differences in flavor and volatile composition between organic and conventional pac choi may be more evident at early stages of growth.

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 (MScAgric)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Wine is a product of the fermentation of grape juice. Alcoholic fermentation is mainly conducted by the yeast Saccharomyces cerevisiae which metabolises grape sugars to mainly ethanol, CO2 and glycerol. Aside from these primary fermentation compounds, the yeast also produces many secondary metabolic by-products that are important to wine quality and style. Malolactic fermentation (MLF) is a secondary fermentation that normally occurs after alcoholic fermentation. Lactic acid bacteria (LAB) are responsible for the conversion of malic acid to lactic acid and CO2 during MLF, which is important for wine deacidification and also contributes to microbial stability. Malolactic fermentation and LAB strains can also influence the aroma profile of wines. The main genera associated with this process are Oenococcus, Lactobacillus, Pediococcus and Leuconostoc. Oenococcus oeni is the main species associated with MLF because it is able to survive the harsh physiochemical environment of winemaking. Recently L. plantarum has also been introduced as a commercial MLF starter culture. Research has started to focus on the potential of wine yeast and LAB interactions or combinations to alter the wine aroma profile via the production and/or degradation of aroma compounds. The overriding goal of this study is to unravel the interactions between wine yeast and different LAB strains and their impact on wine aroma and flavour. The first aim was to assess LAB growth during co- and sequential inoculation strategies, the ability to complete MLF and the impact on the production of aroma compounds in combination with two different yeast strains in a medium containing full complement of nitrogen supplementation. Malolactic fermentation was successful in the different inoculation strategies and the bacterial combination (L. plantarum and O. oeni) completed MLF in the shortest time. The impact of the bacterial strains on the modification of aroma compounds was bigger in co- than sequential inoculation. A general increase in total esters (contributing to the fruity character of wines) especially ethyl lactate and ethyl acetate was observed. The production of esters, volatile fatty acids and higher alcohols proved to be dependent on either the yeast strain used and/or the LAB strains used. The second aim of the research was to assess the effect of NH4Cl (ammonium) and amino acids supplementation on yeast and LAB strains (both in co- and sequential inoculation strategies) and the impact on the aroma profile of the fermented must. Fermentations supplemented with ammonia as sole nitrogen source showed the highest total bacterial growth in terms of cell numbers. Malolactic fermentation was completed in the shortest time with O. oeni and the bacterial combination inoculums. The co-inoculated strategies in combination with amino acids supplementation showed the biggest impact on the aroma compound profiles of the different fermentation strategies and bacterial treatments. A general increase in total esters was observed for NH4Cl additions with ethyl lactate and ethyl acetate showing the highest concentrations. The concentration of esters, volatile fatty acids and higher alcohols were strongly influenced by the yeast and the single LAB strains used. The results generated from this study showed that the chemical composition of the fermentation medium and the selection of yeast and LAB strains are important because these factors have an influence on the aroma and flavour profiles of wines. / AFRIKAANSE OPSOMMING: Wyn is die produk van gefermenteerde druiwe. Die gis, Saccharomyces cerevisiae is verantwoordelik vir alkoholiese fermentasies waar druiwe suikers na hoofsaaklik etanol, CO2 en gliserol gemetaboliseer word. Die gis produseer ook sekondêre metaboliete wat ‘n belangrike bydrae lewer tot wynstyl en kwaliteit. Appelmelksuurgisting (AMG) is ‘n sekondêre fermentasie wat gewoonlik na alkoholiese fermentasie plaasvind. Melksuurbakterieë (MSB) speel ‘n sleutel rol in die omskakeling van appelsuur na melksuur en CO2 gedurende AMG. Hierdie fermentasie lei tot ‘n afname in die suurheidsgraad en verbeter die mikrobiese stabiliteit van die wyn. Appelmelksuurgisting en MSB rasse kan die aroma- en geurprofiel van wyne beïnvloed. Die belangrikste genera wat met AMG geassosieer word is Oenococcus, Lactobacillus, Pediococcus en Leuconostoc. Oenococcus oeni is die mees algemene ras wat vir AMG gebruik word omdat dit in uiterste wyn toestande kan oorleef. Mees onlangs is Lactobacillus plantarum as kommersiële aanvangskultuur vir AMG geïdentifiseer. Navorsing het onlangs meer begin fokus op gis en MSB interaksie of kombinasies as ‘n strategie om die aroma profiele van wyne te verander. Die hoofdoel van die studie is om die interaksie tussen wyngiste en verskillende MSB rasse en die effek op die aroma profile van wyne te bestudeer. Die eerste doelwit was om die impak van die twee giste op die groei en AMG vermoeë van MSB gedurende ko- en sekwensiële inokulasie praktyke en die impak op die produksie van aroma komponente, in ‘n medium wat die volledige stikstof aanvullings bevat, te bestudeer. Appelmelksuurgisting was suksesvol in die verskillende inokulasie praktyke en die bakteriese kombinasie (L. plantarum en O. oeni) het AMG in die kortste tyd voltooi. Die impak van die bakteriese rasse op die modifikasie van die aroma komponente was groter met ko- as sekwensiële inokulasies. Daar was ‘n toename in die totale esterkonsentrasies veral in etiellaktaat en etielasetaat. Die produksie van esters, vlugtige vetsure en hoër alkohole word beïnvloed deur die gisras en MSB rasse wat gebruik word. Die tweede doelwit was om die impak van NH4Cl (ammonium) en aminosure aanvullings op die gis- en MSB rasse gedurende ko- en sekwensiële inokulasie strategieë te bepaal. Melksuurbakterieë se groei was beter met die ammonium aanvulling. Appelmelksuurgisting was in die kortste tyd voltooi met O. oeni en die bakteriese kombinasie. Die ko-inokulasie praktyke in kombinasie met die kompleks aminosure aanvulling het die grootste impak op die produksie van aroma komponente gehad. Daar was weereens ‘n toename in die totale esterkonsentrasies vir die NH4Cl aanvulling, veral in etiellaktaat en etielasetaat. Die gis en MSB rasse speel ‘n rol by die produksie en konsentrasies van esters, vlugtige vetsure en hoër alkohole. Die resultate van hierdie studie bewys dat die chemiese samestelling van die fermentasie medium, die seleksie van gis- en MSB rasse is belangrik omdat hierdie faktore die aroma en geur profiele van wyne beïnvloed.

Les analyses physico-chimiques des arômes du vin prennent aujourd’hui un essor considérable pour faciliter la prise de décision des professionnels de la vigne et du vin. Des analyses performantes, pour un certain nombre de molécules parmi les plus pertinentes, ont été développées (GC-MS, LC-MS). Cependant les techniques utilisées ne sont pas facilement transposables au sein des exploitations. L’objectif de cette thèse est donc de proposer un nouveau type de dosage peu onéreux et simple à mettre en œuvre. La technique envisagée est le dosage immunologique (tests ELISA) qui permet, pour quelques Euros, de doser directement sur le terrain les molécules pertinentes sans préparation préalable des échantillons. Les molécules visées (alkylméthoxypyrazines et précurseurs cystéinylés de thiols volatiles) sont présentes dans les vins de Cabernet Sauvignon et de Sauvignon blanc. / Analytical methods for the detection and quantification of wines aroma typically utilise HPLC-MS or GC-MS. The methods require some isolation and concentration step preceding the analysis. Enzyme-linked immunosorbent assays (ELISAs) are becoming either alternative complementary analytical tools to conventional methods because of their rapidity, sensitivity, selectivity, and low cost. In this Thesis, the applicability of ELISAs for detection and quantification of precursors of volatile thiols and alkylmethoxypyrazines, which have been isolated from wines, made from Cabernet Sauvignon or Sauvignon Blanc, are described.

Ce travail décrit les équilibres gaz-liquide lors des fermentations alcooliques en conditions œnologiques, en se focalisant sur les composés d'arôme les plus abondants : isobutanol, alcool isoamylique, acétate d'éthyle, acétate d'isoamyle et hexanoate d'éthyle. Les coefficients de partage à l'équilibre (ki) de ces molécules sont quantifiés, grâce à la méthode PRV (Phase Ratio Variation), en précisant l'effet de la composition du milieu et de la température. Grâce à la mise en œuvre de fermentations spécifiques pendant lesquelles la vitesse de dégagement de CO2 est contrôlée (grâce à une perfusion d'azote ammoniacal), il a été montré que le dégagement de CO2 n'avait pas d'effet sur le rapport de concentrations entre phase gaz et liquide et pouvait être assimilé au ki. Une démarche de modélisation est ensuite effectuée, pour estimer la valeur du rapport de concentrations entre phase liquide et gaz pour ces composés (à l'exception de l'acétate d'éthyle et de l'alcool isoamylique), à tout moment de la fermentation et quelle que soit la température, y compris en conditions de non isothermie. Le modèle, basé sur l'équilibre des phases gaz et liquide, conduit à une erreur moyenne d'estimation inférieure à 10%. Grâce à ce modèle, il devient possible de réaliser des bilans de production des différents composés tout au long de la fermentation, à partir de leur seule mesure dans la phase gaz. Ces bilans permettent de différencier (i) la quantité globale produite, représentative des potentialités de la levure (intérêt microbiologique), (ii) la quantité restant dans la phase liquide (intérêt organoleptique) et (ii) la quantité perdue dans le gaz effluent (intérêt technologique). Il est ainsi mis en évidence que les pertes d'esters sont très importantes. Par exemple, à 20°C, elles atteignent respectivement 44 % et 25 % pour l'hexanoate d'éthyle et l'acétate d'isoamyle. Grâce à un dispositif de suivi en ligne permettant des mesures très fréquentes (une par heure), les vitesses - et les vitesses spécifiques - de production et de pertes sont calculées. Elles constituent de nouvelles informations utiles à la fois pour l'étude du métabolisme (suivi de la dynamique de flux métaboliques) et pour mieux raisonner les conduites de fermentation, notamment le régime de température. L'impact de ce régime de température est étudié plus en détail en réalisant des bilans de production lors de fermentations conduites avec la même souche de levure et le même moût. Les résultats obtenus montrent que si l'on ne prend en compte que les concentrations dans la phase liquide - généralement les seules informations disponibles - on surestime fortement l'impact de la température sur le métabolisme de composés fortement volatils, tels que les esters / The gas-liquid partitioning during winemaking fermentations was studied, with a focus on the main aroma compounds: isobutanol, isoamyl alcohol, ethyl acetate, isoamyl acetate, ethyl hexanoate. The partition coefficients (ki) of these molecules were quantified, using the PRV (Phase Ratio Variation) method. The influence of the must composition and temperature was assessed. The absence of any effect of CO2 production on gaz-liquid ratio compared to ki value was established by running specific fermentations in which the rate of CO2 production was kept constant by perfusion with assimilable nitrogen. A model was then elaborated to calculate gaz-liquid ratio of these molecules (excepted ethyl acetate and isoamyl alcohol) at any fermentation progress and at any temperature, including anisothermal fermentations. The model based on the equilibrium between the gas and liquid phases predicted ki with less than 10% error. Using this model, balances were c alculated, with a differentiation between (i) the total production, representative of the yeast potential (microbiological interest), (ii) the amount remaining in the fermenting must (organoleptic interest) and (iii) the amount lost in the exhaust CO2 (technological interest). High losses of esters were observed. For example, at 20°C, they represented 44% and 25 % for ethyl hexanoate and isoamyl acetate, respectively. Using an on line monitoring with a high frequency of measurements (one per hour), rates and specific rates of production were calculated. These new data are useful both for studies on metabolism (dynamics of metabolic fluxes) and for improving fermentation control, in particular temperature profile. The impact of temperature was assessed in more detail by comparing balances of production during fermentations run with the same yeast strain and must. These experiments demonstrated that the effect of temperature on the yeast metabolism was highly overestimated whe n only considering the concentrations in the liquid, i.e. usually the only available information.

Des études récentes ont démontré l'existence d’un espace sensoriel typique des vins liquoreux de Bordeaux. Des composés volatils de familles chimiques différentes ont été identifiés et corrélés avec la typicité de ces vins. Cependant, les composés responsables des nuances aromatiques de « fruits confits », considérées comme des nuances « clés » de l’arôme des vins liquoreux de pourriture noble, n’avaient pas fait l’objet de recherches préalables. Les analyses classiques des extraits de vin par CPG-SM n'ont pas permis l’identification des composés responsables des nuances de « fruits confits » et plus précisément des notes d’ « orange confite ». Ainsi, une méthode d’étude alternative, reposant sur des approches sensorielles et instrumentales, a été mis en œuvre afin d’étudier les fractions obtenues par CLHP semi préparative. Des tests d’omission ont été réalisés par l’utilisation de la CPG préparative dans le but d’isoler une fraction contenant l’arôme d’ « orange confite ». Ces tests ont mis en évidence l’impact de cette fraction sur l’arôme des vins puisque son omission, au niveau de l’aromagramme, entraine la perte du caractère « fruit confit » dans les extraits. L’application de la CPG multidimensionnelle, il a permis d’identifier une nouvelle lactone spécifique des vins liquoreux, le 2-nonen-4-olide. Les expériences sensorielles de reconstitution et d’omission, menées à partir de fractions de vin ou en milieu modèle ont permis de souligner l’impact de composés clés - lactones, eugénol et 3-sulfanylhexanol - avec le 2-nonen-4-olide dans l’expression aromatique des nuances de « fruits confits », et plus particulièrement d’ « orange confite » au travers d’un phénomène d’accord aromatique. Par ailleurs, les corrélations entre le niveau de typicité de vins liquoreux de Bordeaux et les teneurs de ces composés ont été étudiées. Enfin, la genèse du 2-nonen-olide a été appréciée au cours de la botrytisation de raisins de Sémillon puis pendant l’élevage des vins en barrique. / Recent studies have demonstrated the existence of a typical sensory concept for Bordeaux dessert wines. Volatile compounds from several chemical families were identified and correlated with aromatic typicality in these wines. However, the compounds responsible for “key” aromas, particularly those recalling “overripe” fruits, had not previously been interpreted. Classical analysis of wine extracts by GC-MS was not successful in identifing the compounds responsible for these ripe fruity notes. An alternative strategy was developed in this research combining both analytical and sensory studies of wine fractions extracts obtained by semi-preparative HPLC. Omission tests were conducted using preparative GC with cryotrapping in order to evidence the fraction presenting overripe fruit nuances focusing on orange notes. These tests demonstrated the impact of this fraction on the wine aroma, as the overripe fruity note disappeared when this section was not trapped. Multidimensional CPG allowed the identification of a new lactone, 2-nonen-4-olide, typical to dessert wines. Reconstitution and omission tests carried out from wine HPLC fractions or in a model wine medium, highlighted the importance of specific compounds - lactones, eugenol and 3-sulfanylhexanol - with 2-nonen-4-olide, on the overripe fruit aromas expression and particularly on orange notes via perceptual blending. Furthermore, the correlations between the level of typicality of Bordeaux sweet wines and the levels of these compounds were studied. Finally, the evolution of 2-nonen-4-olide was evaluated during the botrytisation of Semillon grapes as well as in wines during aging in barrels.

A proposta desse trabalho foi avaliar meios de cultivo para produção de lipase por Penicillium roqueforti e utilizá-la na forma bruta em um processo industrial em escala de bancada de síntese de aroma de queijo, procurando, desta forma, contribuir na busca por processos biotecnológicos viáveis para síntese de aroma natural desse produto que participa da formulação de muitos produtos alimentício, o aroma de queijo. Para tanto, estudou-se a produção de lipase pelo fungo em fermentação submersa, utilizando dez diferentes meios de cultivo, na presença e na ausência de óleo de oliva. Os extratos obtidos foram avaliados quanto à atividade de lipase, protease e quantificação de biomassa. O meio denominado H+óleo induziu a maior atividade lipolítica em 96 h (7,5 U/mL) e foi, portanto, selecionado para aplicação no processo industrial em escala de bancada. A produção da enzima foi realizada utilizando o fungo Penicillium roqueforti em meio H+óleo, sendo esta denominada extrato enzimático bruto, apresentando atividade lipolítica de 10,97 U/mL e atividade específica de 2184,07 U/mg. A atividade proteolítica desse extrato foi 166,30 U/mL e seu perfil eletroforético apresentou 7 bandas proteicas de massas molares variando entre 122,2 kDa e 11,9 kDa. Após a determinação das atividades enzimáticas, este extrato enzimático bruto foi aplicado no processo na indústria através de um planejamento fatorial completo avaliando a influência da temperatura, tempo de reação e pH sobre a intensificação de sabor e odor de queijo, sendo esta determinada através de análise sensorial utilizando Teste de Comparação Múltipla, definindo como padrão o aroma produzido por este processo nas condições realizadas pela empresa. O mesmo planejamento fatorial foi aplicado para este processo avaliando uma enzima comercial. O extrato enzimático aplicado nas condições de temperatura de 50°C, tempo de reação de 24 h e pH 5,0 levou a síntese de um aroma de queijo mais intenso, sendo este o preferido pelos provadores quando avaliado juntamente com o aroma padrão da empresa e o aroma mais intenso resultante do planejamento utilizando a enzima comercial, em análise sensorial utilizando teste de preferência. Esses resultados evidenciam a potencialidade do uso desse extrato enzimático na obtenção de aroma natural de queijo. / Not avaliable.

Orientador: José Roberto Campanha / Banca: Márcio Luiz de Andrade Netto / Banca: Gerson Antonio Santarine / Resumo: Neste trabalho construímos modelos de redes neurais artificiais recorrentes com dois, com quatro, com seis e com oito neurônios na tentativa de simular computacionalmente como os neurônios receptores olfativos dos vertebrados, em especial dos seres humanos, conseguem identificar e reconhecer as diferentes moléculas odoríferas (ou odorantes) transportadas pelo ar. Para isso, usamos uma rede que evolui de um sistema dinâmico caótico, na ausência de odorantes, para o não-caótico, quando do reconhecimento de um odor constituído, no máximo, de até três odorantes. / Abstract: We built models of recurrent artificial neural networks with two, four, six and eight neurons in order to simulate, using computational simulation, the way vertebrate’s olfactory neurons, in special the humans, identify and recognize different odoriferous molecules (or odorants) in the air. For that purpose, we used a network that evolves from a chaotic dynamic system, in the absence of odorants, to the non-chaotic, when it recognizes an odor that is made of, at most, three odorants. / Mestre