The aim of this thesis was to determine the content of biogenic amines (BA)and polyamines (PA)in selected types of cheese and the influence of storage time on the content of biogenic amines.

O gênero Psychotria destaca-se na família RUBIACEAE pela produção de alcalóides bioativos e por sua taxonomia complexa, sendo muitas vezes relacionado aos gêneros Palicourea, Cephaelis, Calycodendron e Calycosia. A divisão de Psychotria nos subgêneros Psychotria, Tetramerae e Heteropsychotria foi proposta com o objetivo de auxiliar a classificação quimiotaxonômica do gênero. Estudos demonstram que o subgênero Psychotria (espécies pantropicais) produz alcalóides poliméricos, formados por duas ou mais unidades de triptamina; enquanto o subgênero Heteropsychotria parece estar envolvido com a produção de alcalóides indol monoterpênicos, de acordo com pesquisas realizadas com diferentes espécies coletadas no Sul do Brasil. O isolamento e purificação dos alcalóides estrictosamida, ácido estrictosidínico e miriantosina, a partir de Psychotria myriantha, corroboram com esta hipótese, permitindo a inclusão da espécie no subgênero Heteropsychotria. A literatura descreve várias atividades para extratos e alcalóides isolados de espécies de Psychotria, como antimicrobiana e analgésica, por exemplo. Neste trabalho, o extrato n-butanólico de alcalóides de P. myriantha, além de seus alcalóides isolados, apresentaram atividade inibidora da migração de leucócitos, sugerindo um efeito antiinflamatório, e capacidade de inibir a ação da enzima acetilcolinesterase. Extratos e alcalóides isolados da espécie foram avaliados quanto à atividade antioxidante em CCD, frente ao DPPH, apresentando resultado negativo. O extrato EBA e o alcalóide ácido estrictosidínico aumentaram o tempo de latência no teste da retirada da cauda frente ao estímulo térmico, indicando uma atividade analgésica do tipo opióide. A influência do ácido estrictosidínico, alcalóide isolado em maior quantidade em massa, sobre os níveis de DA, DOPAC, 3-MT, HVA, 5-HT e 5-HIAA em estruturas cerebrais de ratos foi verificada. Hipocampos de animais que receberam injeção intra-hipocampal bilateral de ácido estrictosidínico (20 μg/μL) apresentaram redução de 83,4 % nos níveis de serotonina, em comparação ao grupo controle, enquanto os córtices desses animais apresentaram redução nos níveis de DOPAC (35,9%), 3-MT (24,7%) e 5-HIAA (9%). Hipocampos e estriados de ratos tratados com injeção i.p. de ácido estrictosidínico (10 mg/kg) demonstraram diminuição de 63,4 e 28,7% nos níveis de 5-HT, respectivamente. As alterações nos níveis de aminas biogênicas nas estruturas avaliadas, além das atividades analgésica e inibidora da acetilcolinesterase, indicam que P. myriantha e espécies do subgênero Heteropsychotria constituem uma potencial fonte de substâncias bioativas no tratamento de distúrbios do sistema nervoso central. / Psychotria genus is an important in RUBIACEAE due to bioactive alkaloids production and complex taxonomy, being related to Palicourea, Cephaelis, Calycodendron and Calycosia genera. The division of Psychotria in Psychotria, Tetramerae and Heteropsychotria subgenera was proposed with the aim of aiding the genus chemotaxonomic classification. Studies demonstrate that Psychotria subgenus produce polyindoline alkaloids formed by two or more triptamine units; whereas Heteropsychotria subgenus seems to be involved with indole monoterpene alkaloids production, according to researches with different species collected in Southern Brazil. Isolation and purification of strictosamide, strictosidinic acid and miriantosine from Psychotria myriantha corroborated this hypothesis, allowing its inclusion into Heteropsychotria subgenus. The scientific literature describes several activities to Psychotria extracts and alkaloids, such as antimicrobial and analgesic, for example. In this work, P. myriantha n-butanolic alkaloid extract and isolated compounds inhibited the leukocyte migration, suggesting an antiinflammatory activity, and a weak ability to inhibit the action of acetylcholinesterase enzyme. Alkaloids and extracts from P. myriantha were evaluated as regards their antioxidant activity using DPPH, with no positive results. EBA and strictosidinic acid increased the latency in the tail flick model, indicating an opioid analgesic activity. The influence of strictosidinic acid on the levels of DA, DOPAC, 3-MT, HVA, 5-HT and 5-HIAA in brain structures of rats was verified. Hippocampus with intra-hippocampal injection of strictosidinic acid (20 μg/μL) displayed a decrease of 83.4% in serotonin levels, in comparison with control group; whereas the cortex showed a decrease in the levels of DOPAC (35.9%), 3-MT (24.7%) and 5-HIAA (9%). Hippocampus and striatum that received intraperitoneal injection of strictosidinic acid (10 mg/kg) showed 5-HT levels reduction of 63.4 e 28.7%, respectively. The biogenic amine levels alterations in the studied structures, associated with the analgesic and acetylcholinesterase inhibitor activities, suggest that P. myriantha and species from the Heteropsychotria subgenus constitute a font of bioactive compounds in the treatment of central nervous system disturbs.

La dénitrification autotrophe utilisant le fer Ferreux est un bioprocédé innovant pour l'élimination des nitrates, en même temps que l'oxydation du fer dans les eaux usées. Les dénitrifiants chimio-autotrophes convertissent le nitrate en azote gazeux et l'oxydation du Fe(II) conduit à la production de précipités de fer ferrique qui peuvent ensuite être enlevés et récupérés. La possibilité de maintenir une dénitrification autotrophe avec le fer ferreux en utilisant une culture mixte de Thiobacillus, un inoculum de boue activée et des cultures pures de la souche Pseudogulbenkiania 2002 et de T. denitrificans dans différentes conditions de pH et d'EDTA:Fe(II) a été initialement étudiée dans des essais biologiques par lots. Des ratios plus faibles d’EDTA:Fe(II) se traduisent par une efficacité et des taux d'élimination des nitrates plus élevés. La culture mixte de Thiobacillus présente le taux d'élimination de nitrate le plus élevé, égal à 1.18 mM•(g VSS•d)-1.Par la suite, la culture mixte de Thiobacillus a été ensemencée dans deux réacteurs à lit tassé à flux montant identiques. Les deux réacteurs (réacteur 1 et 2) ont reçu respectivement 120 et 60 mg / L de nitrate et une alimentation différente de Fe (II) afin de respecter un rapport molaire Fe(II):NO3- de 5:1. L’EDTA a été supplémenté à un rapport molaire EDTA:Fe(II) de 0,5:1. Le pH, le TRH et la température étaient de 6,5-7,0, 31 h et 22 ± 2 ° C. Dans le réacteur 1, le TRH a été raccourci de 31 à 24 h et la concentration de NO3- a été maintenue stable à 250 mg / L. Inversement, le réacteur 2 a été mis en fonctionnement avec un TRH décroissant et une concentration de NO3- en alimentation, maintenant ainsi un taux de charge de NO3- stable. Après environ 80 jours d'incubation, l'élimination des nitrates était de 88% dans le réacteur 1 pour un THR de 31 h. L'élimination de nitrates la plus élevée obtenue dans le réacteur 2 était de 80%. Une diminution du TRH de 31 à 24 h n'a pas affecté l'élimination du nitrate dans le réacteur 1, alors que dans le réacteur 2 l'élimination du nitrate a diminué à 64%.De plus, l'influence des métaux lourds (Ni, Cu, Zn) sur la dénitrification autotrophe utilisant du fer ferreux a été évaluée dans des essais biologiques discontinus, en utilisant les mêmes quatre cultures microbiennes différentes. L'efficacité et les taux d'élimination des nitrates les plus élevés ont été obtenus avec la culture mixte dominante de Thiobacillus, alors que la souche Pseudogulbenkiania de 2002 était la moins efficace. Cu s'est avéré être le métal le plus inhibiteur pour les cultures mixtes. Un impact plus faible a été observé lorsque le Zn a été ajouté. Le Ni présentait l'effet inhibiteur le plus faible. Une sensibilité plus élevée à la toxicité des métaux a été observée pour les cultures pures. Enfin, la caractérisation minérale des précipités obtenus pour les expériences avec du Cu, Ni et Zn a été étudiée. Chez les témoins abiotiques, l'oxydation chimique du Fe (II) a entraîné la formation d'hématite. Un mélange de différents (hydro)oxides de Fe(III) a été observé pour toutes les cultures microbiennes, et en particulier : i) un mélange d'hématite, d'akaganéite et / ou de ferrihydrite a été observé dans les précipités des expériences réalisées avec la culture mixte dominée par la présence de Thiobacillus; ii) en plus d'hématite, de l'akaganeite et / ou de la ferrihydrite, la maghémite a été identifiée lorsque la culture pure de T. denitrificans a été utilisée; iii) l'utilisation de la culture pure de la souche Pseudogulbenkiania 2002 a entraîné la formation d'hématite et de maghémite; enfin, l'enrichissement en boues activées a permis la production d'hématite et de magnétite en plus de la maghémite. Aucune différence concernant la minéralogie des précipités n'a été observée avec l'addition de Cu, alors que l'addition de Ni et de Zn a probablement stimulé la formation de maghémite. Une caractérisation minérale supplémentaire est cependant nécessaire / Ferrous iron mediated autotrophic denitrification is an innovative bioprocess for nitrate removal, simultaneously with iron oxidation in wastewaters. Chemoautotrophic denitrifiers convert nitrate to nitrogen gas and Fe(II) oxidation results in the production of ferric iron precipitates that can be subsequently removed and recovered. The feasibility of maintaining Fe(II)-mediated autotrophic denitrification with a Thiobacillus mixed culture, an activated sludge inoculum and pure cultures of Pseudogulbenkiania strain 2002 and T. denitrificans under different pH and EDTA:Fe(II) conditions was initially investigated in batch bioassays. Lower EDTA: Fe(II) ratios resulted in higher nitrate removal efficiency and rates. The Thiobacillus mixed culture resulted in the highest specific nitrate removal rate, equal to 1.18 mM•(g VSS•d)-1.Subsequently, the Thiobacillus mixed culture was seeded in two identical up-flow packed bed reactors. The two reactors (reactor 1 and 2) were fed with 120 and 60 mg/L of nitrate, respectively, and a different Fe(II) feed in order to respect a molar ratio Fe(II):NO3- 5:1. EDTA was supplemented at a EDTA:Fe(II) molar ratio 0.5:1. The pH, HRT and temperature were 6.5-7.0, 31 h and 22±2°C. In reactor 1, HRT was shortened from 31 to 24 h and NO3- concentration was maintained stable at 250 mg/L. Conversely, reactor 2 was operated with decreasing HRT and feed NO3- concentration, thus maintaining a stable NO3- loading rate. After approximately 80 d of incubation, nitrate removal was 88% in reactor 1 at HRT of 31 h. The highest nitrate removal achieved in reactor 2 was 80%. A HRT decrease from 31 to 24 h did not affect nitrate removal in reactor 1, whereas nitrate removal decreased to 64% in reactor 2.Moreover, the influence of heavy metals (Ni, Cu, Zn) on Fe(II)-mediated autotrophic denitrification was assessed in batch bioassays. The highest nitrate removal efficiency and rates were achieved with the Thiobacillus-dominated mixed culture, whereas Pseudogulbenkiania strain 2002 was the least effective. Cu showed to be the most inhibitory metal for mixed cultures. A lower impact was observed when Zn was supplemented. Ni showed the lowest inhibitory effect. A higher sensitivity to metal toxicity was observed for the pure cultures. Finally, the mineral characterization of the precipitates obtained in the experiments with Cu, Ni and Zn was investigated. In abiotic controls, the chemical Fe(II) oxidation resulted in hematite formation. A mixture of different Fe(III) (hydr)oxides was observed with all microbial cultures, and in particular: i) a mixture of hematite, akaganeite and/or ferrihydrite was observed in the precipitates of the experiments carried out with the Thiobacillus-dominated mixed culture; ii) on top of hematite, akaganeite and/or ferrihydrite, maghemite was identified when the T.denitrificans pure culture was used; iii) the use of the pure culture of Pseudogulbenkiania strain 2002 resulted in hematite and maghemite formation; finally, the activated sludge enrichment allowed the production of hematite and magnetite besides maghemite. No difference in the mineralogy of the precipitates was observed with the addition of Cu, whereas the addition of Ni and Zn likely stimulated the formation of maghemite. Further mineral characterization is however required

The consumption of fermented goods has been related with outbreaks of foods intoxications, like these biogenic amines have been associated to the consummations of these products. The aims went to assess the potential of amines formation in dry fermented sausage added or not with starter and fiber, determination of amines formation in cooked salami added or not with organic acids. The dry fermented sausages were fabricated and cased in artificial collagen no eatable gut then, ripening under controlled conditions of temperature, moisture and air stream. The cooked salamis were added with organic acids (lactic and citric), cased in artificial collagen no eatable gut and cooked. The products were according to the microbiologic available legal patterns. The histamine reached amounts of 145,61; 93,38 and 130,10 mg·kg-1 and the tyramine reached amounts of 67,05; 70,28 and 71,87 mg·kg-1 for the dry fermented sausages treatments: control, starter and starter and fiber respectively. The histamine in the cooked salami treatments: control, lactic acid and citric acid reached amounts of 48,06; 55,37 and 49,16 mg·kg-1 and the tyramine reached amounts of 11,00; 11,06 and 11,35 mg·kg-1 respectively. There were no differences at the level tested (p<0,05). In conclude the starter did not avoid the amine formation, attributed to the polluting bacterium found already in the raw materials. The temperature and the organic acids did not influence in the final amounts of amines in the cooked salami. The taste assess was not influenced by the presence of amines. The addition of glucose and sucrose avoided larger amines amounts in the fermented salami and the wheat fiber did not interfere in the metabolism of the microorganisms. / O consumo de alimentos fermentados tem sido relacionado com surtos de intoxicações alimentares e as aminas biogênicas são associadas ao consumo destes produtos. Os objetivos foram avaliar o potencial de formação de aminas em salame tipo Italiano, adicionado ou não, de cultura starter e fibra; determinar a formação de aminas no salame cozido, adicionado ou não de ácidos orgânicos. Os salames foram elaborados e embutidos em tripa artificial de colágeno não comestível. Maturados sob condições de temperatura, umidade e velocidade do ar controlados. Na fabricação do salame cozido, foram adicionados ácidos orgânicos (láctico e cítrico), embutidos em tripa artificial de colágeno não comestível e as peças cozidas. Os produtos se apresentaram conforme os padrões microbiológicos vigentes. A histamina alcançou valores de 145,61; 93,38 e 130,10 mg·kg-1 e a tiramina valores de 67,05; 70,28 e 71,87 mg·kg-1 para os tratamentos do salame controle, starter e starter e fibra respectivamente. A histamina alcançou valores de 48,06; 55,37 e 49,16 mg·kg-1 e a tiramina valores de 11,00; 11,06 e 11,35 mg·kg-1 para os tratamentos do salame cozido controle, ácido láctico e ácido cítrico respectivamente. Não houve diferenças ao nível testado (p<0,05). Conclui-se que o starter comercial não evitou a formação de aminas, atribuída às bactérias contaminantes encontradas já nas matérias-primas. A temperatura e os ácidos orgânicos não influenciaram nos níveis finais de aminas no salame cozido. A análise sensorial não foi influenciada pela presença das aminas. A adição de glicose e sacarose evitou maiores acúmulos de aminas no salame fermentado e a fibra de trigo não interferiu no metabolismo dos microrganismos.

The main aim of this thesis was to prove the hypothesis that the intake of blood with specific antibodies against sand fly saliva affects sand fly fecundity and mortality. Phlebotomus argentipes and Mesocricetus auratus were used for most experiments. ELISA revealed high levels of specific antibodies in repeatedly exposed hosts. However, a five-day study of mortality and fecundity of bloodfed females demonstrated that feeding on repeatedly bitten hamsters has no effect on number of eggs or survival of females. Salivary antigens of P. argentipes recognized by sera of repeatedly bitten hamsters were characterized by immunoblotting and mass spectrometry. Immunoblotting showed that antibodies in the hamster sera recognize salivary proteins with molecular weight from 25 to 60 kDa. Mass spectrometry revealed that the antigens correspond to D7, apyrases, antigen 5-related proteins and yellow-related proteins. In addition, Phlebotomus females were fed through a chicken membrane on rabbit blood with high concentrations of histamine, serotonin or prostaglandin E2 to find out if they influence fecundity or mortality of sandfly females. While the approximate number of eggs layed by one female did not significantly differ from controls in any experimental group studied, the total mortality of females was lower...

L’objectif principal de cette thèse est de mieux comprendre les différents facteurs contrôlant la pompe biologique de carbone en Atlantique Nord et dans l’Océan Austral, à proximité des îles Kerguelen, en utilisant notamment deux approches: le Thorium-234 (234Th) et le baryum biogénique (Baxs).En Atlantique Nord, les flux d’export de carbone organique particulaire (POC) augmentent lorsqu’ils sont associés à des minéraux biogéniques (silice biogénique et carbonate de calcium) et lithogènes, capable de lester les particules. L’efficacité d’export, généralement plus faible que précédemment supposé (< 10%), est inversement corrélée à la production, soulignant un décalage temporel entre production et export. La plus forte efficacité de transfert, i.e. la fraction de POC atteignant 400m, est reliée à des particules lestées par du carbonate de calcium ou des minéraux lithogènes.Les flux de reminéralisation mésopélagique sont similaires ou parfois supérieurs aux flux d’exports et dépendent de l’intensité du développement phytoplanctonique, de la structure en taille, des communautés phytoplanctoniques et des processus physiques (advection verticale).Comme observé pour le POC, l’export des éléments traces est influencé par les particules lithogènes provenant des marges océaniques, mais aussi des différentes espèces phytoplanctoniques.Dans l’Océan Austral, la zone à proximité de l’île de Kerguelen est naturellement fertilisée en fer, augmentant les flux d’export de fer, d’azote et de silice biogénique. Il a été démontré que la variabilité des flux dépendait des communautés phytoplanctoniques dans la zone fertilisée. / The main objective of this thesis is to improve our understanding of the different controls that affect the oceanic biological carbon pump. Particulate export and remineralization fluxes were investigated using the thorium-234 (234Th) and biogenic barium (Baxs) proxies.In the North Atlantic, the highest particulate organic carbon (POC) export fluxes were associated to biogenic (biogenic silica or calcium carbonate) and lithogenic minerals, ballasting the particles.Export efficiency was generally low (< 10%) and inversely related to primary production, highlighting a phase lag between production and export. The highest transfer efficiencies, i.e. the fraction of POC that reached 400m, were driven by sinking particles ballasted by calcite or lithogenic minerals.The regional variation of mesopelagic remineralization was attributed to changes in bloom intensity, phytoplankton cell size, community structure and physical forcing (downwelling). Carbon remineralization balanced, or even exceeded, POC export, highlighting the impact of mesopelagic remineralization on the biological pump with a near-zero, deep carbon sequestration for spring 2014.Export of trace metals appeared strongly influenced by lithogenic material advected from the margins. However, at open ocean stations not influenced by lithogenic matter, trace metal export rather depended on phytoplankton activity and biomass.A last part of this work focused on export of biogenic silica, particulate nitrogen and iron near the Kerguelen Island. This area is characterized by a natural iron-fertilization that increases export fluxes. Inside the fertilized area, flux variability is related to phytoplankton community composition.

Deep subsurface rock samples from Upper Ordovician strata in the Michigan Basin were analyzed for the presence of microbial communities. High concentrations of biogenic methane were observed in the Upper and Middle Ordovician formations. Total porosity values for the shale, shale hard bed and limestone samples were 7.4%, 2.5% and 1.9%, respectively. Hydrocarbon presence ranged from petroliferous shale, to bituminous layering in shale hard beds, to hydrocarbon odour in limestone. Organic carbon content ranged from 0.5 to 2.5%, highest amount being present in the shale. Environmental DNA was extracted from core samples and PCR amplified using 16S rDNA bacterial primers. PCR performed with archaeal 16S rDNA and methanogen-specific (mcrA) primers did not yield DNA amplification. Gene analysis indicated that bacterial sequences similar to Proteobacteria, Cyanobacteria, Firmicutes, and Actinobacteria were present. Most sequences were not related to known cultivated species. Proteobacteria was the most dominant phyla at all depths and included heterotrophic, lithotrophic, acidophilic, radiotolerant, and sulphate-reducing species of bacteria. This study concludes that the observed biogenic methane is a product of ancient methanogenesis.

Les éponges d'eau douce (Porifera) produisent des spicules siliceuses dont la morphologie a une valeur taxonomique et environnementale. Les sédiments très concentrés en spicules, dénommés spongilite, sont présents en abondance dans le nord-ouest du Minas Gerais (Brésil). Ils se sont formés au cours des derniers 28000 ans. Afin d'étudier l'aptitude de ces dépôts à enregistrer les paléoenvironnements, nous avons procédé, à partir des éponges produites actuellement dans un dans un petit lac du nord-ouest du Minas Gerais (Lagoa Verde), à 2 types de calibrations. (1) Le suivi écologique de M. spinata, éponge unique du lac, a été fait au cours de son cycle de vie annuel. L'éponge produit 4 catégories de spicules en lien avec les variations saisonnières de l'équilibre entre précipitation et évaporation, de la température de l'eau et de la concentration en silicium dissout. La mise en évidence de ces relations conforte les interprétations paléoenvironnementales précédemment tirées des assemblages de spicules fossiles. (2) La composition isotopique en oxygène (δ18O) des silicates biogéniques est communément utilisée comme marqueur paléoenvironnemental. Ce marqueur n'a jamais été calibré pour les spicules d'eau douce. Nous avons testé si M. spinata précipitait ses spicules en équilibre isotopique avec l'eau du lac. La composition δ18O des spicules montre une relation thermo-dépendante positive avec la composition δ18O de l'eau, inverse de ce qui est attendu pour une précipitation à l'équilibre isotopique. Ce résultat suggère un fractionnement cinétique d'origine biologique à déterminer en vue de reconstitutions paléoenvironnementales. / Freshwater sponges (Porifera) produce siliceous spicules with taxonomic and paleoenvironmental value. Sediments with high concentrations of spicules, called spongilite, are present in NW Minas Gerais (Brazil). They formed during the last 28000 years. In order to investigate the reliability of these deposits to record past environmental changes, we proceeded to two kinds of calibration, from sponges currently produced in a small lake of NW Minas Gerais (Lagoa verde). (1) The ecology of M. spinata, the only sponge of the lake, was monitored over its annual cycle. The sponge produced 4 spicules categories in relation with seasonal changes in precipitation, evaporation, water temperature and dissolved silicon content. These relationships confirm previous paleoenvironmental reconstructions from fossil spicule assemblages. (2) The oxygen isotopic composition (δ18O) of biogenic silica is commonly used as a paleoenvironmental proxy. This proxy has never been calibrated for fresh water sponge spicules. We checked whether M. spinata formed its spicules in isotopic equilibrium with the lake water. The δ18O signature of the spicules showed a positive thermo-dependent relationship with the δ18O of the lake water, conversely to what was expected for an equilibrium precipitation. This result suggests that a biological kinetic fractionation occurred. This kinetic fractionation needs to be systematically characterized for paleoenvironmental reconstruction purpose.

Les amines biogènes sont des composés allergènes, notamment rencontrés dans les produits fermentés tel que le vin. Les bactéries lactiques du vin, dont Oenococcus oeni, le principal acteur de la fermentation malolactique, sont capables de produire ces molécules, à partir de précurseurs azotés. Afin de réduire la teneur en amines biogènes, il est nécessaire de comprendre le rôle de cette production, chez les souches impliquées dans la synthèse de ces métabolites. Le projet européen BiamFood FP7 (n°211441) a été mis en place pour répondre à cette problématique. Au cours de mon travail de thèse, des outils moléculaires ont été développés afin de muter de manière ciblée les gènes de O. oeni et d’exprimer des gènes d’intérêt. Ainsi, les clusters hdc de Lactobacillus hilgardii et odc de O. oeni, responsables respectivement de la synthèse de l’histamine et de la putrescine, ont été clonés dans le vecteur pGID052 et transférés dans la souche de laboratoire O. oeni ATCC BAA-1163, déficiente pour la production d’amines biogènes. Cette stratégie n’ayant pas donnée lieu à la production d’amine biogène, mon travail de recherche s’est orienté vers la caractérisation biochimique de l’ornithine décarboxylase de O. oeni. L’ornithine décarboxylase de O. oeni BR14/97 a été sur-produite chez E. coli via le système pET. Après purification de l’enzyme, le pH optimal et la température optimale de fonctionnement, 5,5 et 35°C respectivement, ont été caractérisés. Les constantes cinétiques Km = 1 mM et Vmax = 0,57 U. mg-1 ont également été déterminées en mesurant la putrescine produite par HPLC (Gomez-Alonso et al, 2007). De plus, l’ODC de O. oeni BR14/97 est spécifique de la L-ornithine et ne peut pas dégrader la L-lysine en cadavérine, ce qui implique la présence d’une voie métabolique propre à la production de cadavérine chez cette souche. Dans un second axe du projet, l’influence de la source azotée et notamment des peptides versus acides aminés libres, a été étudiée chez une souche de Lactobacillus plantarum, isolée du vin et productrice de tyramine. Les peptides contenant de la tyrosine, sont utilisés par cette souche pour former de la tyramine. La production augmente en phase stationnaire de croissance, et est corrélée à l’expression du transporteur tyrP. L’analyse de la tyrosine présente dans le milieu de culture montre que ces peptides seraient dégradés dans le milieu extracellulaire chez L. plantarum. / Biogenic amines are indesirable compounds found in fermented products like wine. Lactic acid bacteria from wine, including Oenococcus oeni, the main actor of malolactic fermentation, are able to produce these molecules from nitrogenous precursors. In order to limited biogenic amines accumulation, it is necessary to understand the role of this production by strains responsible for the synthesis of these metabolites in food. That is why the European BiamFood project was put in place. Along my thesis, molecular tools were developed in order to mutate O. oeni genes (encoding decarboxylases), and to express genes of interest. Genetic clusters hdc from L. higardii and odc from O. oeni, responsible for histamine and putrescine synthesis respectively, were cloned into the pGID052 vector and transferred into the laboratory strain O. oeni ATCC BAA-1163, which is not able to produce any biogenic amines. However no biogenic amines production was observed. My work thesis was then redirected in the in vitro biochemical characterization of the ODC from O. oeni. The odc from O. oeni BR14/97 was overproduced in E. coli BL21 via the pET system. Optima pH and temperature were determined with the purified enzyme obtained. Kinetic constants Km = 1 mM and Vmax = 0,57 U.mg-1 were also determined by measuring putrescine production by HPLC (Gomez-Alonso et al, 2007). Furthermore, ODC from O. oeni BR14/97 is specific for the L-ornithine and can not decarboxylated the L-lysine into cadaverine. In a second part of the BiamFood project, the impact of nitrogen sources, especially peptides compared to free amino acids, was studied in a Lactobacillus plantarum strain, isolated from wine and able to produce tyramine. Peptides containing tyrosine residues are used by this strain to produce tyramine. Tyramine production increases during the growth and reaches a maximum at stationary phase. The production is correlated with tyrP expression. The tyrosine measured in culture media shows that peptides may be hydrolyzed in the extracellular medium in L. plantarum.

Orientador: Giuseppina Pace Pereira Lima / Resumo: O mercado de suco de uva está em crescimento, principalmente pelos benefícios do consumo a saúde. Várias soluções tecnológicas foram propostas para melhorar a cadeia produtiva, como a combinação de uvas e até mesmo o aproveitamento do resíduo do processo de produção, contribuindo na qualidade funcional, econômica e ambiental. No Brasil, uvas Vitis labrusca e híbridas representam a maioria daquelas destinadas aos sucos. A uva híbrida 'Niagara Rosada' que apresenta alta produtividade e aceitabilidade, com excedente de produção, pode ser uma opção para a produção de suco e utilização do resíduo para suplementação alimentar. Dessa forma, o perfil de compostos fenólicos e nitrogenados, bem como capacidade antioxidante e bioativa, coloração e sensorial foram realizadas com o objetivo de verificar a qualidade dos sucos de uva e seus resíduos produzidos a partir de combinações de 'Niagara Rosada'. Os sucos foram elaborados com 'Bordo', 'BRS Violeta', 'IAC 138-22 Máximo', separadamente e em blends com 'Niagara Rosada'. Os sucos foram produzidos por prensagem a quente, engarrafados (vidro âmbar) e pasteurizados. Um controle comercial (C) de 'Niagara Rosada' também foi analisado junto aos sucos. Os resíduos da produção do suco (bagaços) foram secos, moídos, embalados e armazenados para as análises. Treze misturas foram produzidas independentemente usando 100, 75, 50 e 25% de cada cultivar e em combinação com 'Niagara Rosada'. O perfil de polifenóis e compostos nitrogenados foram analisa... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The he grape juice market is growing, mainly due to the health benefits of consumption. Various technological solutions have been proposed to improve the production chain, such as the combination of grapes and even the use of the residue of the production process, contributing to functional, economic and environmental quality. In Brazil, Vitis labrusca and hybrid grapes represent the majority of those destined for juices. The hybrid grape 'Niagara Rosada', which presents high productivity and acceptability, with surplus production, can be an option for the production of juice and use of the residue for food supplementation. Thus, the profile of phenolic and nitrogen compounds, as well as antioxidant and bioactive capacity, coloring and sensory were carried out with the aim of verifying the quality of grape juices and their residues produced from 'Niagara Rosada' combinations. The juices were made with 'Bordo', 'BRS Violeta', 'IAC 138-22 Máximo', separately and in blends with 'Niagara Rosada'. The juices were produced by hot pressing, bottled (amber glass) and pasteurized. A commercial control (C) of 'Niagara Rosada' was also analyzed with the juices. The juice production residues (bagasse) were dried, ground, packaged and stored for analysis. Thirteen mixtures were produced independently using 100, 75, 50 and 25% of each cultivar and in combination with 'Niagara Rosada'. The profile of polyphenols and nitrogen compounds were analyzed via HPLC. Total phenolic compounds, total ... (Complete abstract click electronic access below) / Doutor