Aroma is one of the most important attributes involved in wine quality. Current trend in winemaking consists of producing wines with different aroma nuances to offer variety of wines to a developing market. Several studies have demonstrated that low temperature fermentations favours aroma synthesis and retention. In this background, new wine yeasts able to perform fermentation at low temperatures improving wine aroma while maintaining good fermentation rates are necessary. This doctoral thesis explores the oenological traits of different Saccharomyces species and hybrids relevant for present-day wine industry, especially regarding aroma production, as well as the molecular bases underneath. This exploration has been possible using different biochemical, analytical chemistry and molecular techniques to perform enzymatic activity detection, aroma profile determination and transcriptome analysis in wine fermentations. Through this doctoral thesis the abilities of different Saccharomyces species and hybrids regarding primary aroma release and secondary aroma production, especially at low temperatures, has been elucidated in order to know the different possibilities that these yeasts offer to create new wines with different aromatic nuances. One of the general conclusions of this doctoral thesis is that production and release of aromas in winemaking depends on the strain carrying out the fermentation process. Nevertheless, sometimes there was a species tendency. On the other hand, the fact that fermentation temperature affects aroma synthesis but not always in the direction to aroma increase has been demonstrated. / Gamero Lluna, A. (2011). Study of the production and release of aromas during winemaking carried out by different Saccharomyces species and hybrids [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/10741

Omega-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA, C20:5, n-3) and docosahexaenoic acid (DHA, C22:6, n-3), have many medically established benefits against cardiovascular diseases, cancers, schizophrenia, and Alzheimer's. Currently, fish oil is the main source of omega-3 fatty acids, but there are many problems associated with it such as undesirable taste and odor, and heavy metal contamination. As a result, it is necessary to seek alternative production sources based on various microorganisms. In this thesis we have developed a novel microfungal culture process to produce EPA from the crude glycerol byproduct generated in biodiesel industry. This process provides both an alternative source of omega-3 fatty acids and a benefit to the biodiesel industry. Indeed, as oil prices reach historical highs, biodiesel has attracted increasing interest throughout the United States. The disposal of the crude glycerol byproduct has been a challenge faced by the biodiesel producers. Crude glycerol presents a cheap carbon source for growth of many microorganisms. In this thesis, we tested the feasibility of using crude glycerol for producing eicosapentaenoic acid (EPA, 20:5, n-3) by one algal species, Phaeodactylum tricornutum and two fungal species, Mortierella alpina and Pythium irregulare. We observed that the algal growth is inhibited in the crude glycerol while the fungi can grow very well in crude glycerol-containing medium. The fungus M. alpina produced significant amount of ARA but negligible amount of EPA. P. irregulare produced significant amount of biomass as well as a relatively high level of EPA. The maximum dry biomass for the P. irregulare culture was 2.9 g/L with an EPA productivity of 7.99 mg/L-day. Based on these results, we concluded that P. irregulare was a promising candidate for EPA production from biodiesel derived crude glycerol. Further optimization work showed that P. irregulare grown 30 g/L crude glycerol and 10g/L yeast extract results in the highest level of EPA production. A temperature of 20o C is optimal for high fungal biomass and EPA levels. Addition of vegetable oil (at 1%) enhanced the EPA production and almost doubled the amount of biomass reached. Soap inhibits growth as well as EPA production severely even in small amounts. Methanol completely inhibits growth. The final optimized growth conditions for the fungus P.irregulare were a medium with 30g/L of crude glycerol, 10 g/L of yeast extract at a pH of 6 with 1% supplementation of oil, at a temperature of 20o C for a period of 7 days.Thus we have established that the fungus P.irregulare can be used successfully to produce high mounts of EPA from crude glycerol. / Master of Science

No

Le perméat de lactosérum (PL) et le son de blé (SB) sont des coproduits de l’industrie laitière et céréalière hautement disponibles et généralement utilisés pour l’alimentation animale ou l’épandage. Dans un contexte d’économie circulaire, des bio-ingrédients durables et innovateurs peuvent être produits par la valorisation des co-produits afin de répondre à la demande de produits avec des bienfaits santé par les consommateurs. Par ailleurs, la fermentation est une alternative de recyclage biologique qui permet d’augmenter la disponibilité des composés bioactifs du SB et de produire des exopolysaccharides à partir du PL. Dans ce contexte, l’objectif de cette étude est d’obtenir un bio-ingrédient (BI) riche en polysaccharides et composés bioactifs par fermentation du mélange de PL/SB par trois souches de Lacticaseibacillus rhamnosus R0011, ATCC 9595 et RW-9595M en monoculture ou en coculture avec Saccharomyces cerevisiae. Dans un premier temps, les expériences ont visé l’évaluation de l’effet de la coculture sur la production d’exopolysaccharides (EPS) par les souches de L. rhamnosus dans du PL. La croissance des mono et cocultures en milieu liquide a été suivie par le compte microbiologique (CFU/mL), la viabilité (PMA-qPCR), la consommation du substrat (lactose) et la production des métabolites (acide lactique et EPS) pendant 48h sans contrôle de pH. L’analyse transcriptomique par RT-qPCR a été utilisée pour déterminer l’expression relative des gènes en coculture par rapport à la monoculture au cours de la fermentation. Un total de 34 gènes en lien avec l’opéron EPS, le métabolisme des sucres, le stress et l’environnement ont été ciblés. Pour l’obtention du bio-ingrédient le mélange de PL /SB a été fermenté en milieu solide par trois souches bactériennes en mono et coculture. L’évolution de la fermentation a été suivie par comptage microbiologique (CFU/g), par la consommation du lactose, le suivi de l’acidification, la détermination des protéines solubles, r la rhéologie et par microscopie électronique à balayage.Le milieu fermenté a été lyophilisé pour obtenir les BI. La composition des BI, le contenu en polysaccharides hydrosolubles (PSH) qui incluent les EPS, la capacité antioxydante et la bioaccessibilité in vitro des composés phénoliques totaux et des acides phénoliques ont été comparés avec le son de blé non fermenté (SBNF). Dans du PL, après 48 heures, les résultats ont révélé que la production d’EPS était améliorée de 39%, 49% et 42% en coculture pour R0011, ATCC 9595 et RW-9595M, respectivement. Des taux d’expression de l’opéron de EPS plus élevés ont été observés pour la souche la plus productrice d’EPS (RW-9595M) en coculture. La construction de réseaux de co-expression de gènes a révélé des corrélations communes entre l’expression de gènes liée aux opérons EPS, le métabolisme du sucre et le stress au cours de la production des EPS et la croissance des trois souches. Les propriétés des BI montrent que le mélange PL/SB c’est un produit riche en protéines, composés phénoliques, fibre alimentaire et polysaccharides solubles dont la quantité est rehaussée en présence de la levure principalement pour les souches ATCC 9595 et R0011. En général, les produits fermentés ont une teneur en composés phénoliques (TPC) liée et une activité antioxydante supérieure au SBNF. Cette différence est plus accentuée en monoculture par rapport à la coculture dans les différents BI. Le TPC total et l'activité antioxydante étaient 30% et 21% plus élevés en monoculture pour le BI obtenu avecRW-9595M par rapport à SBNF. Après la digestion in vitro, la bioaccessibilité des acides phénoliques totaux libres a été améliorée de plus de 40 % dans les BI par rapport au SBNF. Par la fermentation, on observe un enrichissemment en acide caféique libre hautement bioaccessible (superieur à 75%) dans les BI résultant de l’action des bactéries lactiques, ce qui n’a jamais été publié dans la littérature. Les BI obtenus en coculture par les souches RW-9595M et R0011 montrent un contenu significativement plus élevé en acides phénoliques liés qui diminuent lors de la digestion in vitro par rapport à la monoulture. Nos résultats donnent un aperçu de l’influence positive de l’interaction bactérie lactique-levure sur la stimulation IV de la biosynthèse des EPS et les propriétés bioactives du PL/SB fermenté, ce qui représente un progrès important dans la mise au point d’un procédé pour l’obtention d’un bio-ingrédient avec de vastes applications industrielles. / Whey permeate (PL) and wheat bran (SB) are by-products from the dairy and cereal industries that are highly available and used for animal feed or spreading. In a circular economy, sustainable and innovative bio-ingredients can be produced through the valorization of food by-products to be able to respond to the demand for health products from consumers. In addition, fermentation is an alternative to biological recycling to increase the availability of bioactive compounds from SB and to produce exopolysaccharides from PL. In this context, the objective of this study is to obtain a bio-ingredient (BI) rich in polysaccharides and bioactive compounds by fermentation of the mixture of PL / SB by three strains of Lacticaseibacillus rhamnosus R0011, ATCC 9595 and RW-9595M in monoculture or in coculture with Saccharomyces cerevisiae. Initially, the experiments aimed at evaluating the effect of coculture on the production of exopolysaccharides (EPS) by the L. rhamnosus strains in PL. The growth of mono and coculture s in liquid medium was followed by the microbiological account (CFU / mL), the viability (PMA-qPCR), consumption of the substrate consumption (lactose) and the production of metabolite productions (lactic acid and EPS) during 48 h without pH control. Transcriptomic analysis by RT-qPCR was used to determine the relative expression of genes in coculture compared to monoculture during fermentation. In total 34 genes linked to the EPS operon, sugar metabolism, stress and the environment were targeted. To obtain the bio-ingredient, the PL / SB mixture was fermented in solid medium by the bacterial strains in mono and coculture. The progress of the fermentation progress was followed by microbiological account (CFU / g), by the consumption of lactose consumption, by the monitoring of acidification, the determination of soluble protein determinations, by rheology and by scanning electron microscopy. At the end, the fermented medium was lyophilized to obtain the BI. The composition of BI composition, the content of water-soluble polysaccharides content (WSP) which include EPS, the antioxidant capacity and the in vitro bioaccessibility of total phenolic compounds and phenolic acids were compared with unfermented wheat bran (UFWB). In PL, after 48 hours, the results revealed that EPS production was improved by 39%, 49% and 42% in coculture for R0011, ATCC 9595 and RW-9595M, respectively. Higher EPS operon expression rates were observed for the more EPS-producing strain (RW-9595M) in coculture. The construction of gene co-expression networks has revealed common correlations between gene expression linked to EPS operons, sugar metabolism and stress during the production of EPS production and the growth of the three strains. The properties of BI properties show that the fermented SB/PL mixture is rich in protein, phenolic compounds, dietary fiber and soluble polysaccharides, the amount of which is enhanced in the presence of yeast mainly for ATCC 9595 and R0011. In general, fermented products have a content of bound phenolic compounds (TPC) and an antioxidant activity greater than UFWB. This difference is more accentuated in monoculture compared to coculture in for different BI. Total TPC and antioxidant activity were 30% and 21% higher in monoculture for BI obtained from RW-9595M compared to UFWB. After in vitro digestion, the bioaccessibility of total free phenolic acids was improved by more than 40% in BI compared to UFWB. Through fermentation, we observe an enrichment in highly bioaccessible free caffeic acid (superior than 75%) in BI resulting from the action of lactic acid bacteria, which was not reported before in the literature. The BI obtained in coculture with RW-9595M and R0011 have a significantly higher content of bound phenolic acids which decrease during in vitro digestion compared to monoculture. Our results provide an overview of the positive influence of the lactic acid bacteria-yeast interaction on the stimulation of EPS biosynthesis and the bioactive properties of fermented PL / SB, which represents major progress in the development of a process for obtaining a bio-ingredient with wide industrial applications.

Ruminant livestock are a significant contributor to global greenhouse gas emissions, accounting for 6% of total emissions worldwide. Over the past two decades, extensive research has examined the environmental impact of methane (CH4) emissions from these animals. Recent scientific interest has focused on seaweed as a potential CH4 inhibitor, with the compound bromoform identified as the key active component responsible for reducing CH4. A review of the literature, and different mitigation strategies are mentioned in chapter 1 of this dissertation.The second chapter of this dissertation discussed the effects of different bromoform concentration levels on enteric CH4 production and rumen fermentation using 24 h rumen batch culture systems. The doses tested ranged from 0.02 to 13.4 μl bromoform per liter of rumen fluid. The ANKOM batch culture system was utilized, with samples collected after 24 h of incubation for analysis of volatile fatty acids (VFAs) and CH4 levels. Statistical analysis using one-way ANOVA, Student's T-test, and Dunnett's model in JMP Pro revealed a linear reduction in CH4 production as bromoform concentrations increased from 0.02 to 13.4 μl/L (P<0.001). Total CH4 mitigation occurred at doses from 0.20 to 13.4 μl/L (P<0.01), while lower doses of 0.02, 0.05, and 0.10 μl/L reduced CH4 production by 4, 8, and 23% respectively compared to the control (P<0.001). Total VFA concentrations responded quadratically to bromoform treatment. High doses (1.67-13.4 μl/L) caused a reduction in total VFAs (tVFAs), whereas low doses (0.02-0.10 μl/L) had no significant effect. Decreased tVFAs at high doses corresponded with reductions in the molar proportions of acetate and propionate and an increased acetate: propionate ratio (P<0.001). Conversely, low doses decreased the acetate: propionate ratio (P<0.001). Butyrate molar proportions increased across all dose levels (P<0.01).On Chapter 3, we investigated the long-term effects of different doses of bromoform, an active compound from the red seaweed Asparagopsis taxiformis, on CH4 production and rumen fermentation characteristics using a continuous fermenter system. Four treatments were tested - control (no bromoform) and 0.14, 0.27, 0.56 μl/L bromoform doses - in a Latin square design with four 10-day periods. Bromoform doses were divided and dosed with a 50:50 forage: concentrate diet three times daily. Results showed bromoform doses up to 0.56 μl/L reduced enteric CH4 production by 98% compared to control, while increasing hydrogen (H2) concentrations by 550-860% and decreasing ammonia (NH3-N) and pH. Total VFA and dry matter digestibility (DMD) were unaffected, but bromoform altered individual VFA proportions, decreasing acetate, iso-butyrate and acetate: propionate ratio while increasing propionate and butyrate.The withdrawal effect of bromoform was investigated in chapter 4 using four continuous fermenters over two 10-day periods. Treatments were control with no bromoform (T1), bromoform (0.14 μl/L rumen fluid/day) added for the first 5 days (T2), 7 days (T3), or all 10 days (T4) of each period. Bromoform addition reduced CH4 concentrations by 98% and H2 levels compared to control, but these effects nearly disappeared 3-5 days after bromoform dosing ended. The molar proportions of acetate decreased while the molar proportions of propionate increased with bromoform, though these changes were less pronounced 3-5 days post-dosing. Total VFA, NH3-N, and DMD were unaffected by treatment diets. The last chapter of this dissertation discusses the use of Laser Methane Detector (LMD) and Optical Gas Imaging (OGI) for detecting and quantifying enteric CH₄ emissions from ruminants in vitro. Four dietary treatments with varying forage-to-concentrate ratios(F:C) were tested in rumen fermenters. The control treatment (T1) was a 50:50 forage to concentrate diet (F:C), while T2 was the control diet with bromoform added, a known CH4 inhibitor, T3 was a high forage diet (80:20 F:C), and T4 was a low forage diet (20:80 F:C). LMD measurements aligned with the standard gas chromatography (GC) method. Results showed that diet composition significantly impacts CH₄ production: the high-forage diet (T3) produced the most CH₄, while adding bromoform fully inhibited emissions. OGI images analyzed with semantic segmentation models, particularly Gasformer, effectively detected CH₄ plumes. The study concluded that combining OGI with deep learning models can effectively monitor CH₄ emissions to inform dietary and management practices to mitigate climate change. In conclusion, bromoform significantly mitigated CH4 emissions without significantly impairing rumen fermentation however, the effect of bromoform can be reversed with the discontinuation of the treatment suggesting its mode of action is unlikely related to altering the rumen microbial profile. Future research is necessary to evaluate the effect of adding bromoform on feed intake and animal health, and the different aspects of introducing bromoform to the animal feeding industry

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.

<p>Promene komponenata i strukture mleka ispitane su tokom fermentacije kori&scaron;ćenjem inokuluma kombuhe kultivisane na crnom čaju zaslađenom saharozom<br />u koncentraciji 10%. Fermentacija mleka sa 2,2% mlečne masti izvr&scaron;ena je na dve različite temperature (37&deg;C i 42&deg;C) i uzorci su analizirani na sledećim pH vrednostima: 6,1; 5,8; 5,4; 5,1; 4,8 i 4,6. Urađena je determinacija i identifikacija produkata fermentacije laktoze kao &scaron;to su: &scaron;ećeri, organske kiseline i masne kiseline. Ispitane su teksturalne i reolo&scaron;ke karakteristike i mikrostruktura uzoraka. Pored toga, utvrđene su sličnosti i razlike uticaja inokuluma kombuhe na proces fermentacije mleka u poređenju sa delovanjem jogurtne, odnosno probiotske starter kulture.<br />Tokom fermentacije mleka u različitim fazama procesa pri odabranim pH vrednostima na 42&deg;C transformi&scaron;e se 14,6% laktoze odnosno 18,2% na 37&deg;C. Sadržaj galaktoze i glukoze koje nastaju fermentacijom i hidrolizom laktoze raste između prve i druge tačke fermentacije (pH=6,07 i pH=5,8). Dominantne organske kiseline tokom fermentacije su L&ndash;mlečna kiselina i sirćetna kiselina. Nije utvrđena značajna razlika u sadržaju masnih kiselina tokom fermentacije mleka kombuhom na 37&deg;C i 42&deg;C. Sadržaj palmitinske kiseline u mleku i uzorcima tokom fermentacije je najveći, zatim slede miristinska, stearinska i oleinska kiselina.<br />Praćenjem procesa fermentacije mleka uz primenu kombuhe na 37&deg;C i 42&deg;C od početne pH vrednosti 6,07 do zavr&scaron;ne 4,6, najznačajnije promene teksturalnih karakteristika (čvrstoće, konzistencije, kohezivnosti i indeksa viskoziteta) i reolo&scaron;kih svojstava zabeležene su između pH=5,4 i 5,1, &scaron;to je u korelaciji sa mikrostrukturom.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Promene viskoziteta u svim uzorcima pokazuju istu regresionu liniju, sa različitim koeficijentima i visokom vredno&scaron;ću r2, osim uzorka proizvedenog primenom jogurtne kulture na pH 5,4. Uzorak dobijen kori&scaron;ćenjem inokuluma kombuhe imao je najveće vrednosti kompleksnog modula, &scaron;to je rezultiralo boljim reolo&scaron;kim karakteristikama gotovog proizvoda.<br />Da bi se definisao empirijski model procesa fermentacije laktoze u mleku delovanjem kombuhe formulisana su prethodno dva matematička modela za kinetiku fermentacije saharoze u tradicionalnom kombuha napitku (na crnom čaju) &ndash; jedan koji opisuje promenu koncentracije saharoze tokom fermentacije, i drugi koji opisuje brzinu fermentacije. Promena koncentracije laktoze na ispitivanim temperaturama 37&deg;C i 42&deg;C sastoji se od dve lag faze između kojih je faza izraženog pada koncentracije. Krive zasićenja pokazuju sigmoidalnu kinetiku na nižim koncentracijama laktoze, ukazujući na kompleksni ne&ndash;Michaelis&ndash;Mentenov tip kinetike.<br />Generalno može se istaći da su primenom inokuluma kombuhe tokom fermentacije mleka u različitim fazama procesa ustanovljene specifične promene komponenata i strukture u poređenju sa efektima delovanja jogurtne odnosno probiotske starter kulture.</p><p>&nbsp;</p> / <p>Changes of milk components and structure were examined during fermentation by kombucha inoculum cultivated on black tea switened with saccharose in a concentration of 10%. The fermentation of milk with 2.2% of milk fat was carried out at two different temperatures (37&deg;C i 42&deg;C) and samples were analyzed at the following pH values: 6.1; 5.8; 5,4; 5.1; 4.8 and 4.6. Determination and identification of the lactose fermentation products, such as: sugars, organic acids and fatty acids were carried out. Textural and rheological characteristics and microstructure of the samples were investigated. Furthermore, the effect of kombucha on the milk fermentation process was compared with the effect of yoghurt and probiotic starter culture.<br />During milk fermentation at various stages of the process 14.6% of lactose content was transformed at 42&deg;C and 18.2% at 37&deg; C. The galactose and glucose content, which are formed by lactose hydrolyses and fermentation, increased between first and second pH point of fermentation (pH = 6.07 and pH = 5.8). Dominant organic acids during fermentation are L&ndash;lactic acid and acetic acid. There is no significant difference between samples in fatty acids content during fermentation on 37&deg;C and 42&deg;C. The level of palmitic acid in milk and samples was the highest of all fatty acids, followed by myristic, stearic, and oleic acid.<br />The most significant changes in textural properties (firmness, consistency, cohesiveness and viscosity index) and viscosity during milk fermentation by kombucha at 37&ordm;C and 42&deg;C, from the initial pH value 6.07 to a final 4.6, were recorded between pH=5.4 and 5.1, which is in correlation with microstructure.<br />Viscosity changes in all samples showed the same regression line with the different coefficients and a rather high r2 except for the sample produced with standard yoghurt culture at the pH 5.4. Samples produced with kombucha had the highest values of the complex modulus, which indicates better rheological characteristics of the final product.<br />In order to define an empirical model of lactose fermentation process in milk by kombucha, two mathematical models (one for the change of saccharose concentration, during its fermentation by kombucha, and the other for the rate of the mentioned fermentation) previously were formulated. Change of lactose concentrations at 37&ordm;C and 42&ordm;C consists of two retaining stages and very steep descend in-between. Saturation curves show a sigmoidal kinetics at low lactose concentrations, indicating a complex non&ndash;Michaelis&ndash;Menten type kinetics.<br />Generally it can be concluded that specific changes in the components and structure of milk by application of kombucha inoculum during fermentation at different stages of the process were established in comparison with the effects of yoghurt and probiotic starter culture.</p>

Les activités anthropiques entraînent une contamination des sédiments de rivière en de nombreux polluants et en particulier en éléments traces métalliques (ETM). Si la majorité des ETM se retrouvent piégés dans les sédiments, ceux-ci peuvent être remobilisés et passer en solution dans certaines conditions physico-chimiques et sous l’action des microorganismes autochtones. Les métaux relargués peuvent alors constituer un danger potentiel pour les organismes vivants dans les sédiments et dans la colonne d'eau. Dans le cas des sols, l’impact de l’activité microbienne autochtone sur la mobilité des ETM a souvent été rapporté. Cependant une telle activité de solubilisation n’a été que rarement étudiée dans le cas des sédiments. Une telle connaissance est pourtant importante pour la prédiction du comportement des métaux contenus dans les sédiments et la gestion de ces derniers, notamment lors de leur stockage suite aux opérations de dragage. Dans ce contexte, l’objectif de cette thèse a été de comprendre et d’évaluer l’importance de certains des processus microbiens et chimiques de mobilité des ETM dans les sédiments en conditions anaérobies. La première phase de notre étude qui a consisté à incuber des sédiments de Seine et de Marne en milieu anaérobie dopé en glucose avait pour objectif d’étudier la corrélation entre le métabolisme microbien et le comportement des métaux en solution et dans les sédiments. Dans ces conditions opératoires, une forte solubilisation du fer et du manganèse (sous forme réduite) associée à une solubilisation de métaux traces (Co, Cu, Ni) a été mise en évidence, ce qui a laissé supposer l’intervention de bactéries ferri-réductrices dans les phénomènes observés. Une activité fermentaire importante a été observée et caractérisée par la production d’acides organiques majoritaires tels que les acides acétique et butyrique. Un tel résultat souligne l’importance des bactéries fermentatrices dans les phénomènes de dissolution observés. La deuxième étape de ce travail a consisté à confirmer l’importance de l’activité ferri-réductrice et à en identifier les acteurs principaux. Les analyses moléculaires menées ont montré que les bactéries ferri-réductrices majoritairement identifiées, appartiennent aux espèces Clostridium butyricum et Paenibacillus polymyxa. L’utilisation d’un modèle géochimique nous a permis de montrer que les voies métaboliques supportant la réduction du fer et la mobilité des métaux étaient les fermentations butyrique et acétique. La troisième étape a consisté à comparer les impacts directs (réduction enzymatique) et indirects (propriétés des acides organiques produits) de Paenibacillus polymyxa et Clostridium butyricum sur la mobilité du fer, du manganèse et des autres métaux. Une telle étude a montré que les acides organiques produits (acétique, lactique, succinique, propionique et butyrique) ont un très faible impact sur la solubilisation aux pH rencontrés dans les sédiments et que la réduction enzymatique microbienne est le principal mécanisme de dissolution des éléments métalliques en milieu anaérobie / Antropic activities lead to the metallic contamination of river sediments. Most of trace metals are sorbed on sediments but a part of them can be released into aquatic environment when environmental conditions are modified. This is often due by the autochthonous microbial activity. Microbial activites and their consequences on the mobility of metals have been widely studied in soils. Metals are released through direct or indirect microbial mechanisms. Such studies in the case of sediments are very seldom. However, it can be usefull to understand the microbial mechanisms of metal release in sediments, and particularly for a good management of dredged sediments. In this environmental framework, the aim of this research work was to understand and to evaluate the role of the microbial and chemical mechanisms in the release of metals from river sediments in anaerobic conditions. Firstly, sediments from the Marne and Seine rivers were incubated in anaerobic conditions. A high solubilisation of iron and manganese occurred associated to the solubilisation of trace metals (Co, Cu, Ni, Pb). Meanwhile, organic acids were produced and the medium was acidified. Thus fermentation was supposed to be the main process of microbial metabolism. Furthermore these observations led us to suppose the presence of iron-reducing bacteria. In a second step, the extent of the iron-reducing activity was studied. The main iron-reducing bacteria identified in the Marne sediments belonged to the species Clostridium butyricum and Paenibacillus polymyxa. The use of a geochemical model revealed that fermentation and reduction of iron(III) were the main metabolic pathways. Finally direct (enzymatic reduction) and indirect (complexation with organic acids, acidification) impacts of iron-reducing bacteria on the release of metals were compared. Acidification and organic acids had a weak impact on metal solubility in the range of studied pH (between 6,5 and 5). Enzymatic reduction is the main mechanisms of metal release in anaerobic conditions. Indeed the metallic concentrations can be 40 times higher in the presence of iron-reducing bacteria

Parce que les ressources fossiles sont épuisables par définition, le carbone nécessaire à la production d'énergie et de matériaux pourrait provenir en grande partie de la biomasse lignocellulosique. Les procédés de fermentation sont capables de fournir une grande variété de produits d'intérêts capables de remplacer les synthons d'origine pétrolière. Cependant, en raison (i) de son caractère insoluble, (ii) de sa structure plus ou moins cristalline et (iii) de la nature des liaisons entre les maillons du polymère, la cellulose est un substrat carboné difficile à valoriser par voie biochimique/fermentaire seule. La pyrolyse rapide ou la liquéfaction de la cellulose sont principalement étudiées pour produire une bio-huile, qui serait valorisée par hydrotraitement catalytique en carburant ou en building blocks. Dans l'état de l'art actuel, les travaux à l'interface de ces deux domaines portant sur une conversion biochimique ou microbiologique de ces bio-huiles sont encore rares. L’objectif de cette thèse est de coupler un procédé de conversion thermochimique de la cellulose, pour la dépolymériser, à un procédé de transformation microbienne pour produire des solvants, des acides et des gaz (butanol, éthanol, acétone, acide acétique, acide butyrique, acide lactique, hydrogène) qui suscitent un fort intérêt dans l’industrie des carburants ou de la chimie verte. Pour ce faire, le bois de hêtre a été fractionné par les méthodes organosolv et chlorite/acide (SC/AA) afin de récupérer une pâte riche en cellulose. Des procédés de liquéfaction hydrothermale et de pyrolyse rapide ont été utilisés pour obtenir des sucres qui ont été finalement transformés par fermentation en synthons. De nombreuses méthodes analytiques ont été développées pour la caractérisation des produits issus de chaque étape du procédé. Enfin, un modèle du procédé utilisant le logiciel commercial Aspen Plus® a été développé pour établir les bilans de matière et énergie du procédé intégré : du fractionnement du bois, puis la liquéfaction de la fraction cellulosique et à la fermentation des bio-huiles / Because fossil resources are exhaustible by definition, the carbon needed for energy and materials production could be obtained from lignocellulosic biomass. Fermentation processes are able to provide a wide variety of interesting products that can replace the crude oil based "building blocks". However, the abundance of lignocellulosic biomass in the environment contrasts with its very low bioavailability. Indeed, because of (i) its insoluble nature, (ii) its more or less crystalline structure and (iii) the nature of the bonds between the polymer fibers, cellulose is a carbon substrate difficult to valorize by biochemical/fermentation processes alone. Fast pyrolysis or liquefaction of cellulose are mainly studied to produce a bio-oil, which would be upgraded by catalytic hydrotreatment into fuels or building blocks. In the current state of the art, studies at the interface of these two fields involving a biochemical or microbiological conversion of these bio-oils are still rare. The aim of this thesis is the coupling of a thermochemical conversion process of cellulose, to depolymerize it, to a microbial transformation process to produce solvents, acids and gases (butanol, ethanol, acetone, acetic acid, butyric acid, lactic acid, hydrogen) that are of great interest for the fuel or green chemistry industry. To do this, beech wood was fractionated by organosolv and chlorite / acid (SC / AA) methods in order to recover a cellulose-rich pulp. Hydrothermal liquefaction and fast pyrolysis processes were used to obtain sugars that were transformed into building blocks by fermentation. Many analytical methods have been developed for the characterization of products from each step of the process. Finally, a model of the process using the commercial software Aspen Plus® was developed to establish mass and energy balances of the integrated process including: the fractionation of the wood, then the liquefaction of the cellulosic fraction and the fermentation of bio-oils

Moët & Chandon est le leader mondial incontesté dans le secteur des vins mousseux de haute qualité. La sélection des différentes gammes de vins de qualité se fait sur la base de la dégustation de vin, après la fermentation malolactique par la dégustation des jurys d'experts. Une partie importante du volume vinifié ne peut être classé parmi les vins les plus qualitatifs à cause d'un caractère organoleptique appelé «trait lactique». Cette caractéristique est due à la formation de métabolites volatils au cours de la fermentation du vin en particulier diacétyle, acétoïne et de 2,3-pentanedione. Le diacétyle, également connu sous le nom de 2,3-butanedione, est le principal marqueur associé à l'apparition de notes beurrées. Moët & Chandon souhaite commencer une thèse pour contrôler l'apparition de trait lactique dans ses vins de bases. Bien que la production de diacétyle se produit à la fois pendant la fermentation alcoolique (FA) et la fermentation malolactique, la thèse se concentrera sur la gestion de la FA. Une approche multi-factorielle basée à la fois sur l'ingénierie microbiologique et chimique sera mis en œuvre. Dans un premier temps, le travail visera à mieux comprendre le fonctionnement et la régulation des voies métaboliques impliquées dans la formation de diacétyle. De cette information, nous allons développer des stratégies de contrôle de la fermentation afin de minimiser le caractère lactique de vins. Dans ce travail, le rôle et l'impact des paramètres de fermentation sera particulièrement évalué. Le projet commencera par la mise en œuvre des outils d'analyse nécessaires au suivi des marqueurs de trait lactique (diacétyle, acétoïne, 2,3-pentanedione, ...). Deuxièmement, sur un support synthétique, le projet permettra de caractériser le processus de fermentation. Surtout, les équilibres azotés seront contrôlés et les cinétiques de synthèse des composés responsables de trait d'arôme lactique seront déterminées. Une étude de l'impact des acides aminés précurseurs permettra de mieux comprendre le rôle du métabolisme de l'azote. Enfin, l'étude de l'effet de divers facteurs tels que la nature des substances nutritives, température de fermentation et l'apport en oxygène devrait conduire à l'élaboration de stratégies de contrôle rendant possible la diminution du caractère lactique dans les vins destinés à la fermentation secondaire. Dans la dernière partie de projet, les différentes stratégies développées sur moûts synthétiques seront évaluées dans des conditions de vinification réelles. / Champagne company Moët & Chandon is the undisputed worldwide leader in the high quality sparkling wines sector. The selection of different ranges of quality wine is done on the basis of the wine tasting, after malolactic fermentation by tasting expert juries. A significant part of the vinified volume cannot be valued among the most qualitative wines because of a particular feature called « lactic trait ». This trait is attributable to the formation of volatile metabolites during wine fermentation particularly diacetyl, acetoin and 2,3-pentanedione. Diacetyl, also known as 2,3-butanedione, is the main marker associated with the appearance of milky-buttery notes. Champagne company Moët & Chandon wishes to initiate a thesis to control the onset of lactic trait in its wines. Although the production of diacetyl occurs both during alcoholic fermentation (AF) and malolactic fermentation, the thesis will focus on the management of AF. A multi-factorial approach based on both microbiological and chemical engineering will be implemented. At first, the work will aim to better understand the functioning and regulation of metabolic pathways involved in the formation of diacetyl. From this information, we will develop some strategies of fermentation control to minimize the lactic trait of wines. In this work, the role of fermentation parameters will be particularly evaluated. The project will start by the implementation of the analytical tools needed to monitor markers of lactic trait (diacetyl, acetoin, 2,3-pentanedione, ...). Secondly, on synthetic media, the project will allow to characterize the fermentation process. Especially, nitrogen balances will be performed and the kinetics of synthesis of the aroma compounds responsible of lactic trait will be determined. A study of the impact of the precursor amino acids will enable to better understand the role of the nitrogen metabolism. Finally, the study of the effect of various factors such as the nature of nitrogen nutrients, fermentation temperature and oxygen intake should lead to the development of control strategies making possible the decrease of the lactic trait in wines intended for the secondary fermentation. In the final part of the work, the different strategies developed on synthetic must will be assessed in real winemaking conditions.