La fabrication fromagère a depuis longtemps réalisé sa révolution technologique avec la disparition progressive des techniques fermières et l'avènement des pratiques industrielles. Les cuves de cuivre ont été délaissées au profit du tout inox et l'utilisation du lait pasteurisé a augmenté. Néanmoins une part non négligeable des micro-organismes d'affinage provident encore d'un ensemencement spontané par le lait, l'environnement ou le matériel de fabrication. Cette flore naturelle est en grande partie à l'origine : (i) de la richesse et de la diversité organoleptiques des produits traditionnels, (ii) des différences observées entre un fromage industriel fabriqué avec du lait pasteurisé réensemencé avec un levain standard et un fromage " Apellation d'Origine Contrôlée au lait cru " de qualité. Compte tenu de l'évolution des pratiques de fabrication (renforcement des règles d'hygiène réduisant les sources naturelles d'ensemencement traitement du lait par pasteurisation ou microfiltration ; exigence de reproductibilité en utilisant des souches spécifiques présélectionnées), l'utilisation de flores d'affinage sous forme de ferments à additionner au lait doit être de plus en plus envisagée. Le secteur " artisanal " a un grand besoin de recherche et de développement (R&D) afin de conserver ses fabrications traditionnelles, comme les A.O.C qui représentent plus de 10% de la production fromagère en valeur. Les professionnels du fromage souhaitent identifier les souches les plus appropriées à chacun de leurs produits, afin de garantir leur typicité et une reproductibilité de fabrication. Les fromages à pâte molle et croûte lavée comme le Munter et l'Epoisses possèdent une croûte, appelée morge, allant du beige au brun, en passant par le jaune et le rouge-orangé. C'est cette couleur, en plus du caractère organoleptique particulier, que les fromagers cherchent à typer et à reproduire par l'utilisation des souches pigmentées. On a longtemps pensé que cette pigmentation était due uniquement au " ferment du rouge ", Brevibacterium linens. Mais des études récentes ont mis en évidence le rôle non négligeable d'autres bactéries sur la pigmentation de ces fromages, en particulier les corynébactéries et les microcoques. La recherche et la sélection de bactéries pigmentées sauvages par des méthodes simple à mettre en œuvre, la connaissance des mécanismes microbiologiques et moléculaires de la synthèse de pigments et de la coloration des fromages, permettraient de développer des cocktails de souches spécifiques à chaque production fromagère. La première phase du projet consistera en l'isolement de plusieurs centaines de souches sur des fromages issus de terroirs suivants : Epoisses, Reblochon, Munster, Livarot. Ensuite, pour chaque A.O.C., 25 souches d'Arthrobacter correspondant à des teintes variables seront caractérisées d'un point de vue pigmentation :- aspect, teinte sur milieu gélosés,- cinétiques de production en milieu solide et en milieu liquide,- cultures en masse, méthodes d'extraction des pigments,- quantité de pigments produite (production volumique mg/L de milieu) (production spécifique mg/g MS),- valeurs spectrocolorimétriques (L a*b*C*h),- profil HPLC,- HPLC-MS,- RMN des pigments purifies. Les facteurs influençant la production de pigments (substrats ; cultures mixtes-levures désacidifiantes / bactéries ; lumière…) seront étudiés, en portant l'accent sur des milieux de type fromage, voire des caillés modèles. / Smear-ripened soft cheeses, characterized by their orange-red color on rind, are dairy products widely consumed in Europe. The surface color is due essentially to carotenoids, in combination with other pigments, produced by the cheese microflora during ripening. Arthrobacter sp. is one of the major microorganisms occurred on the surface of cheeses, particularly in smear-ripened cheeses, where it is assumed to be responsible for yellow pigmentation of the cheese rind because of its characteristic overall color and its involvement at the different stages of cheese ripening. Pigment-producing microorganisms are commonly found in the nature. Nowadays, pigment-producing microorganisms have been increasing of interest in many scientific disciplines and applications have broadened in the industry because of their biotechnological advantages. As the present trend entirely the world is shifting toward the use of eco and biodegradable products, the requirement for natural ingredients, especially natural colorants, is increasing day by day. The first part of this thesis highlights the crucial role of microorganisms as potential sources of natural pigment production by reviewing a large number of research works related to pigments biosynthesized by microorganisms which were published over the past 10 years by private companies or academic laboratories, with an emphasis on pigments providing for the application in foods. Since the genus Arthrobacter is a group of metabolically versatile bacteria which widely distributed in nature, some parts of this thesis include the review presenting the possibility to produce pigmented Arthrobacter sp. biomasses as novel sources of food colorants; furthermore, the beneficial aspects of Arthrobacter sp. and their promising significances in the dairy industry are also addressed. Considering the significance of Arthrobacter sp. in smear-ripened cheeses, the economically important dairy products, the aim of research described in this thesis is to investigate the implication of this bacterium, particularly Arthrobacter arilaitensis, in the coloration of these cheeses in several aspects covering (i) diversity of pigment production among strains, (ii) kinetic of pigment synthesis, (iii) identification of chemical characteristic of pigments, (iv) colorimetric characterization of pigmentation, and (v) influences of environment i.e. light, pH, NaCl and deacidifying yeasts on the production and the color development of pigments. Among 14 strains of Arthrobacter arilaitensis studied, two groups depending on their ability of carotenoid production could be divided, carotenoid-producing and non-pigmented strains. A growth-associated pigmentation probably applied to indicate the kinetic of carotenoid synthesis by these strains. The diversity of pigment concentration among the carotenoid-producing strains was low, related to the characteristics of pigmentation determined by quantitative spectrocolorimetry. The HPLC-PDA-APCI-MS analysis of extracted pigments of a representative strains revealed 8 different carotenoids showing C50 decaprenoxanthin as the major accumulated carotenoids. Changes in the color development of A.arilaitensis strains under the influences of physical, chemical and biological factors were obtained through spectrocolorymetry. Three groups depending on a coloration behavior affected by light were illustrated e.g. positively sensitive, negatively sensitive and not sensitive to light. The acidic pH and high concentration of salt showed the efficiency inhibited effect on pigmentation of a representative strain of carotenoid-producing A. arilaitensis. In combination of pH and NaCl, deacidifying yeasts were obviously related to the pigment production of A. arilaitensis. The highest average value of color saturation were observed on the studied media deacidified by Debaryomyces hansenii at pH 7.0, displaying intense yellow.

Arthrobacter belongs to an underexplored genus of antimicrobial-producing bacteria. All species in this genus are Gram-positive and commonly isolated from soil and marine sources. This genus is known to exhibit metabolic versatility greatly influenced by environmental conditions and nutrition. Arthrobacter sp. strain TAJX1902 isolated from an unknown soil sample is shown to inhibit a filamentous indicator type bacterium. It produced a few bioactive secondary metabolites in rich medium (RM) broth and agar culture. TAJX1902 crude extract was primarily fractionated using a Sephedex LH-20 column. Further purification achieved via Flash Column Chromatography and Preparative Thin layer Chromatography yielded five prominent crude products. The scope of this project is to fully determine the structure of potential novel antibacterial compounds from TAJX1902 via spectroscopic techniques including full 2D-NMR spectroscopic data set in combination with High-Resolution Mass Spectroscopy (HRMS). The research work is currently underway towards the final purification of the compounds using High-Performance Liquid Chromatography (HPLC) method to enable structural determination and evaluation of their antimicrobial activities.

No description available.

La dissipation des Hydrocarbures Aromatiques Polycycliques (HAP), polluants persistants majoritaires des sols de friches industrielles, implique l'action de microorganismes bactériens et fongiques. Cependant, leur contribution relative à la dissipation in situ, en fonction des interactions entre ces microorganismes ou avec les plantes et les polluants, est mal connue et les communautés fongiques ont été très peu étudiées dans de tels environnements. L'objectif de cette thèse était de préciser l'écodynamique des HAP dans la rhizosphère sous l'effet des microorganismes et de leurs interactions et d'évaluer la diversité fongique dans des sols contaminés. La dynamique des communautés fongiques a été étudiée in situ pendant 5 années, par PCR en temps réel et Temporal Temperature Gradient Electrophoresis (TTGE), dans un sol historiquement contaminé en présence ou non de plantes et dans le même sol ayant subi un traitement de remédiation par désorption thermique. Les plantes avaient un effet positif sur l'abondance et la diversité des champignons et étaient le facteur déterminant la structure des communautés fongiques, dominées par les Ascomycètes. D'autre part, des souches bactériennes et fongiques ont été isolées de ce sol et testées pour leur capacité à dégrader les HAP in vitro. Parmi celles-ci, la bactérie Arthrobacter oxydans Pyr2MsHM11 et le champignon Fusarium solani MM1 ont été choisies comme souches modèles pour étudier leur action individuelle ou conjointe sur la dissipation de trois HAP, dans différentes conditions, des plus simples (cultures liquides) aux plus complexes (microcosmes de sol planté en présence d'une microflore). Les résultats ont montré l'importance fondamentale des interactions entre microorganismes, notamment des phénomènes de compétition, pour la croissance des souches modèles et pour l'expression de leurs potentiels de dissipation des HAP / The dissipation of Polycyclic Aromatic Hydrocarbons (PAHs), very common and persistant pollutants in soils from industrial wastelands, involve the action of bacterial and fungal microorganisms. However their respective contribution, and the influence of the microbial and plant-microbe interactions on in situ PAH dissipation, are poorly known and fungal communities were scarcely studied in such environments. This work aimed to study the fate of PAHs in rhizosphere under the influence of microorganisms and their interactions and to estimate the fungal diversity in contaminated soils. The dynamic of fungal communities was monitored in situ for 5 years, by real-time PCR and Temporal Temperature Gradient Electrophoresis (TTGE) in an aged PAH-polluted soil and in the same soil treated by thermal desorption. The results showed that plants had a positive effect on fungal abundance and diversity and were the main driver of fungal community structure, dominated by Ascomycetes. Besides, bacterial and fungal strains were isolated from this soil and screened for their ability to dissipate PAHs in vitro. Among them, the bacteria Arthrobacter oxydans Pyr2MsHM11 and the fungus Fusarium solani MM1 were chosen as model strains to study their individual and simultaneous effect on PAH dissipation in different experimental conditions, from liquid cultures to planted soil microcosms with a complex microflora. It was found that interactions between microorganisms, notably competition, had a crucial influence on their growth and on the expression of their PAH dissipation potential

North American petroleum refineries use landfarming to dispose of hydrocarbon-containing wastes for bioremediation by indigenous soil microorganisms. In this study, we isolated PAH-degrading bacteria from landfarm soil by enrichment with hydrocarbon-containing effluent. One isolate, Arthrobacter sp. UG50, was capable of using phenanthrene and anthracene as sole carbon sources. The strain degraded phenanthrene (200 mg/L) within 24 h in pure culture at high cell density (10e8 cells/mL). Anthracene (50 mg/L) was slowly degraded, with 29% degraded within 21 days. The strain could not use naphthalene, pyrene, chrysene or benzo(a)pyrene as sole carbon sources, but could degrade pyrene (50 mg/L) cometabolically when phenanthrene was provided. Anthracene degradation (50 mg/L) was enhanced by phenanthrene, with 100% degraded within 6 days. The addition of strain UG50 to petroleum sludge in baffled flasks increased total hydrocarbon degradation and degradation of low concentrations of fluorene, phenanthrene, pyrene and chrysene compared to flasks with limited aeration or containing sludge alone. / NOVA Chemicals and the Ontario Centres of Excellence

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 4 uppsatser.

Universiẗat, Diss., 2001--Stuttgart.

This work is aimed for Actinobacteria and describes their importance in Arctic soil. The members of Actinobacteria are known for their ability to decompose complex natural biopolymers and because they are able to live in harsh arctic environment they could play there an important role in organic matter decomposition. The work compares their abundance in different soil horizons with the focus on cryoturbations and determines the influence of temperature on their amount. This work also represents the results of testing pure Actinobacterial isolates for the production of cellulolytic enzymes.

As antimicrobial resistance persistently disrupts the treatment of microbial infection, natural product chemistry has played a significant role in identifying novel drugs with novel modes of action critical to getting ahead of resistance. The primary goal of this project is to extract and identify potential novel antimicrobial natural products produced by Arthrobacter sp. Although underexplored, Arthrobacter sps. have been known to produce bioactive compounds with versatility; one such is a depsipeptide with quorum-sensing inhibitory activity.1 In this research, an Arthrobacter sp. TAJX1902, isolated from a soil sample, showed inhibitory activity against a filamentous indicator bacterium and a violacein-producing Janthinobacterium sp. Arthrobacter sp. TAJX1902 was cultured using rich medium broth and agar to extract and isolate metabolites. Isolated compounds were characterized via spectroscopic techniques, including 1D and 2D-NMR spectroscopy and GCMS analysis. Arthrobacter sp. TAJX1902 produces Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(phenylmethyl), and five other bioactive cyclic dipeptides (CDPs).

京都大学 / 新制・課程博士 / 博士(農学) / 甲第23530号 / 農博第2477号 / 新制||農||1088(附属図書館) / 学位論文||R3||N5361(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 矢﨑 一史, 教授 小川 順, 教授 阪井 康能 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM