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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
151

Development of Ceramic Thin Films for High Temperature Fiber Optic Sensors

Jiang, Hongmin 24 September 2013 (has links)
No description available.
152

Assessment of new catalysts for electrochemical reduction of carbon dioxide

Goel, Ekta 09 August 2019 (has links)
The industrial revolution caused the release of carbon dioxide (CO2) into the atmosphere leading to a climate crisis. The impact of more CO2 in the atmosphere has been experienced by everybody. The summers are longer and hotter, while the winters are colder and shorter. The ocean water has become more acidic threatening the ocean life. There is an immediate need to reduce CO2 and switch to alternate energy for human survival. Electrochemical reduction of CO2 (ERC) is a promising technology capable of converting excess CO2 into valueded products. The process of recycling CO2 can address the problem of excess CO2 and is a sustainable solution until our dependence on fossil fuels is reduced. However, currently there are very few catalysts that can convert CO2 into valuable products with a low overpotential. The current research evaluates new catalysts for their ERC potential. [Ni(cyclam)]2+ is a well-known catalyst used to reduce CO2 homogeneously. Therefore, it was used as a standard to optimize the CO2 evaluation protocol. Two new catalysts developed in Dr. Hollis's laboratory, a Pt- pincer and a Fepincer molecule were assessed using this method. Cyclic voltammetry and bulkelectrolysis (BE) experiments were performed under Ar and CO2 environments. The gaseous products from BE were primarily CO and H2 and their quantitative measurement was performed using gas chromatography. Formate determination was performed using UV-Vis spectroscopy. Faradaic yields were calculated for CO, H2, and formate. The overpotentials were calculated for all the processes, and a comparison was made to determine the most efficient process. The turnover numbers (TON) and the turnover frequencies (TOF) of all the catalysts were calculated. Based on all the criteria, the Fepincer complex was determined to be the most promising catalyst for further optimization. Additionally, a Faradaic efficiency calculation spreadsheet was created to improve calculation efficiency. The protocol described here has been successfully applied to assess new catalysts and can prove to be an invaluable tool when numerous catalysts require evaluation.
153

CO<sub>2</sub>-selective Membranes for Fuel Cell H<sub>2</sub> Purification and Flue Gas CO<sub>2</sub> Capture: From Lab Scale to Field Testing

Salim, Witopo 01 June 2018 (has links)
No description available.
154

Characterization of the H10/A4 Region of Vesicular Stromatitis Virus G Protein and Effects of H2-H10/A4 Mutations of Fusogenic Functions / VSV G H10/A4 Mutants and H2-H10/A4 Double Mutants

Shokralla, Shahira 11 1900 (has links)
The vesicular stomatitis virus glycoprotein G is responsible for low pH mediated membrane fusion induced by the virus. Four linker insertion mutants (H2, H5, HIO, A4) of the G ectodomain were found to disrupt fusion and yet maintained all the requirements for proper folding and cell surface expression (Li et al., 1993). Site specific mutagenesis of residues 123 to 137, surrounding the H2 mutant, either blocked or shifted the pH optima and threshold of fusion to more acidic values with a concomitant reduction in cell-cell fusion efficiency (Zhang and Ghosh, 1994; Fredericksen and Whitt, 1995). The region is highly conserved among vesiculoviruses and was found to insert into lipid membranes by hydrophobic photolabelling (Durrer et al., 1995) suggesting a possible role for this domain as the fusion peptide. Site-directed mutagenesis of residues 190 to 210, surrounding the H5 insertion mutant, did not significantly affect fusion (Fredericksen and Whitt, 1995). Surrounding the H10 and A4 insertion mutants is a conserved region, residues 395 to 424, that does not interact with target membranes (Durrer et al., 1995). To determine the functional importance of this region, site-directed mutagenesis was employed. Substitution of conserved Gly 404, Gly 406, Asp 409, and Asp 411 with Ala, Ala, Asn, and Asn, respt:.ctively, both reduced fusion and caused a shift in the pH of fusion threshold to more acidic values (tested by Y. He as published in Shokralla et al., 1998). In this study, the Hl0/A4 region is further mutagenized and tested for fusion. Cell surface expression was examined by indirect immunofluorescence and lactoperoxidase catalyzed iodination. Rates of transport from the endoplasmic reticulum and oligomerization into trimers were tested by resistance to endoglycosidase H and sucrose density gradient centrifugation, respectively. Low-pH induced conformational changes were assayed by resistance to proteolytic digestion. Residues Gly 395, Gly 404, Gly 409 and Ala 418 were substituted with Glu, Lys, Asp, and Lys, respectively. All mutants, with the exception of A418K, were expressed at levels similar to or above wild-type. Mutants G404K and D409A completely abolished fusion. Mutant G395E reduced cell-cell fusion efficiency by 82% and shifted both the pH threshold and optimum of wild type fusion. Although all mutants were capable of trimer formation, alterations in the structure of mutants G404K, D409 A, and A418K were detected by slower transport rates. All Hl0/A4 mutants were more susceptible to trypsin than wild-tyr,e at the pH of6.5, and mutant G404K was completely susceptible at this pH Reductions in the extent of fusion, along with shifts in the pH optima and thresholds of fusion suggest that the Hl0/A4 region (residues 395 to 418) of vesicular stomatitis virus G protein is important for G mediated fusion. The region may influence low-pH induced conformational changes. Double mutants of the H2 and HI0/A4 regions were also tested for their effects on fusion. The extents of fusion mediated by double mutant G proteins were severely reduced with levels ranging from 28% wild-type fusion to complete fusion deficiency. Only mutant Gl31A G404A was capable of 83% wild-type fusion. Mutants Gl31A G395E, Gl31A G404A, Gl31A D4LIN, Dl37N G404A, and the fusion defective D137N D411N were expressed at levels above wild-type G protein at the cell surface. Mutants Fl25Y D411N and Pl26L D411N, although capable of very low levels of fusion were not detectable at the cell surface by immunoflorescence and were detected at low levels by lactoperoxidase catalyzed iodination of cell surface proteins. These two mutants, along with Gl31A G404A, also showed slower transport rates than wild-type G. All double mutants showed increased sensitivity to trypsin at the pH of 6.5 with mutant Fl25Y D411N showing complete susceptibility. They were also all capable of trimer formation by sucrose density gradient centrifugation. In comparing the fusion profiles of double mutants with those of their component single mutants, it was found that in most cases the pH threshold of fusion by double mutants was greater than the sum of the single mutants and that the pH optimum of fusion corresponded to that of the constituent H2 single mutant. Although, the regions are functionally independent, they may indirectly affect one another through alterations in protein structure. / Thesis / Master of Science (MS)
155

A sustainable integration approach of chlor-alkali industries for the production of PVC and clean fuel hydrogen: prospects and Bangladesh perspectives

Roy, H., Barua, S., Ahmed, T., Mehnaz, F., Islam, M.S., Mujtaba, Iqbal M. 22 August 2022 (has links)
Yes / The chlor-alkali industries produce caustic soda (NaOH), chlorine (Cl2 ), and hydrogen (H2 ) as primary products. In 2021, the global chlor-alkali market was valued at $63.2 billion. The article evaluates the global aspects of chlor-alkali industries and prospects for Bangladesh. The current production capacity of NaOH from the chlor-alkali industries in Bangladesh is around 282,150 metric tons/year (MT/y). The by-products, chlorine (Cl2 ) of 250,470 MT/y and hydrogen (H2 ) of 7055 MT/y, are produced domestically. The local demand of Cl2 is 68,779 MT/y. However, there are no systematic utilizations of the residual Cl2 and vented H2 , which threatens the sustainability of the chlor-alkali industries. The article prefigures that a 150,000 MT/y PVC plant can utilize 45.2 % of residual Cl2 of chlor-alkali plants, which would be an economical and environmental milestone for Bangladesh. The residual Cl2 can earn revenue of 908 million USD/y, which can be utilized to import ethylene. For the sustainable utilization of vented H2 , production of H2O2 , fuel cell electric vehicle (FCEV) and H2 fuel-cell-based power plant are the feasible solutions. Thus, for the long-term growth of the chlor-alkali industry in Bangladesh and other developing countries, systematic utilization of Cl2 and H2 is the only feasible solution. / This research was funded by ‘BUET Chemical Engineering Forum (BCEF), 001-2020.
156

Briquetting and Reduction of DRI/HBI Fines for Next-Generation Iron and Steel Making Technology

NITUNGA, Eddy-Nestor January 2024 (has links)
The current transition from Blast Furnace (BF) to Hydrogen-Direct Reduction (H-DR) is an initiative to reduce the carbon footprint in Iron and Steel production. The shift from traditional coke-based Iron and Steel making to a green H-DR requires new innovative technologies, underscoring the importance of collective efforts in this field. The production of DRI/HBI fines, which account for 1-2% wt. of DR production, poses a significant challenge for the iron and steel industries. These fines, rich in iron, are generated inside the plant during the handling, production, and transportation process. Their recycling in the DR process is difficult without proper agglomeration. Here, briquetting emerges as a promising solution to this challenge. This research work is dedicated to exploring the use of innovative binders in briquetting the DRI/HBI fines. It aims to understand the H2 -based reduction behaviour of the briquettes and the role of embedded biocarbon in enhancing resource efficiency. The study also investigates the impact of binders on the mechanical strength, moisture content, and compaction pressure of the briquettes. The optimized briquettes are then subjected to H2-based reduction using a thermogravimetric technique (TGA) followed by characterization by XRF, XRD, and LECO analysis, and mechanically evaluated to assess their potential in the next-generation H2-based Iron and Steel Making. Keywords: DRI/HBI Fines, Organic binders, Briquetting, H2 Reduction, Biocarbon, Recycling, Resource Efficiency
157

Contrôle actif de l'accélération latérale perçue d'un véhicule automobile étroit et inclinable / Active lateral acceleration control of a narrow tilting vehicle

Mourad, Lama 19 December 2012 (has links)
Les Véhicules Etroits et Inclinables (VEI) sont la convergence d’une voiture et d’un motocycle. Un mètre de largeur seulement suffit pour transporter une ou deux personnes en Tandem. Les VEI sont conçus dans le but de résoudre partiellement les problèmes de trafic routier, de minimiser la consommation énergétique et l’émission de polluants. De par leurs dimensions(ratio hauteur/largeur), ces véhicules doivent s’incliner en virage pour rester stable en compensant l’effet de l’accélération latérale. Cette inclinaison doit dans certains cas être automatique : elle peut être réalisée à l’aide d’un couple d’inclinaison généré par un actionneur dédié (système DTC), soit encore en modulant l’angle de braquage des roues (Système STC). Nous avons proposé dans ce mémoire une méthodologie de synthèse d’un régulateur structuré minimisant la norme H2 d’un problème bien posé au bénéfice d’une régulation optimisée de l’accélération latérale, considérant tour à tour les systèmes DTC et STC. Les régulateurs proposés sont paramétrés par la vitesse longitudinale et s’avèrent performants et robustes, et les moyens de réglages proposés permettent d’étudier l’intérêt relatif d’une solution DTC pure ou mixte DTC/STC, permettant de supporter les développements futurs sur le sujet. L’originalité des solutions proposées en regard des études rencontrées dans la littérature porte en particulier sur le fait de choisir de réguler directement l’accélération latérale perçue (plutôt que l’angle d’inclinaison), en anticipant la prise de virage par la prise en compte des angles et vitesse de braquage. L’optimisation de la régulation permet de réduire de manière importante le couple d’inclinaison requis, et l’accélération latérale subie par les passagers est faible. Tous les développements proposés s’appuient naturellement en amont sur un travail de modélisation (recherche du modèle juste nécessaire), et de bibliographie conséquent. Le modèle retenu comprend 5 degrés de libertés. Nous avons démontré qu’il possédait la propriété intéressante d’être plat, et avons utilisé cette propriété pour ouvrir des perspectives relatives à la conception d’un régulateur non-linéaire robuste, susceptible apriori d’accroître les performances dans le cas de « grands mouvements ». Au contraire de ce qui existe dans la littérature,le régulateur multivariable conçu pour le système SDTC permet le contrôle coordonné des actions sur les systèmes STC et DTC. / Narrow Tilting Vehicles (NTV) are the convergence of a car and a motorcycle. One meter wide, these vehicles are designed for one or two people sitting the one in front the other. The idea behind the conception of NTV is the minimization of traffic congestion, energy consumption and pollutant emission. But because of their dimensions, these cars would have to lean into corners in order to compensate for the lateral acceleration and maintain their stability. The tilting should be automatic, and can be achieved by a tilting torque generated by a dedicated tilting actuator (DTC) or by modifying the steering angle (STC) or both (SDTC). In this thesis, we first propose a methodology for the design of an output feedback structured regulator, minimizing the H2 norm of a well-posed problem, built to optimize the lateral acceleration of the NTV, considering DTC and SDTC systems.The designed controllers, with the longitudinal velocity as a parameter, lead to the minimization of the tilting torque and of the lateral acceleration perceived by the driver, and have good performances as well as good robustness properties. Furthermore, the tuning methodology allows the comparison of a pure DTC solution and a mixed SDTC alternative. Compared to the literature, the originalities in this thesis are the direct control of the measured value of the lateral acceleration (instead of the tilting angle), and the anticipation of the tilt, thanks to the use of the steering angle and angular velocity. Furthermore, the SDTC solution allows to drive both the STC and DTC systems in a coordinated manner. The design strategies are based on a preliminary study of vehicle models, and a design model with 5DoF was developed. We demonstrated that the model has the nice property to be flat, and in the last section of the thesis, used this property to initiate the design of a non-linear robust controller, which can a priori lead to better performances in case of “large motions”.
158

Estudo farmacognóstico comparativo de Passiflora alata Curtis e Passiflora nítida Kunth (Passifloraceae). Avaliação das atividades antiúlcera e antioxidante dos seus extratos / Comparative pharmacognostic study of Passiflora alata Curtis and P. nitida Kunth (Passifloraceae). Evaluation of antiulcer and antioxidant activities of its extracts

Wasicky, André 15 October 2007 (has links)
Passiflora alata Curtis e P. nitida Kunth, espécies brasileiras, foram submetidas ao estudo farmacognóstico comparativo. P. alata tem sido utilizada há tempos na medicina tradicional e em preparações farmacêuticas. No estudo farmacobotânico, as folhas das duas espécies apresentaram semelhança na forma, no tamanho e na ausência de indumento. Diferem quanto ao número de glândulas peciolares. P. alata apresenta geralmente dois pares de glândulas e P. nitida, um par. Anatomicamente, as duas espécies mostraram características comuns ao gênero Passiflora: mesofilo dorsiventral; drusas no mesofilo, na nervura mediana, na região cortical, medular e floemática do caule; feixes vasculares colaterais. Embora as duas espécies apresentem nervura mediana biconvexa, a de P. alata evidencia as carenas pronunciadamente salientes, principalmente na face abaxial. Outro aspecto diferencial observa-se na forma do caule: em P. alata é quadrangular e, em P. nitida, arredondada. A seqüência de tecidos assemelha-se, com exceção do maior desenvolvimento de colênquima nas arestas de P. alata. A triagem fitoquímica evidenciou a presença de flavonóides nas duas espécies e a ausência de alcalóides em ambas, tanto na droga vegetal quanto nos extratos. Formação de espuma persistente foi observada com a droga vegetal preparada com P. alata. A hemólise foi observada com a droga e o extrato desta espécie. Os flavonóides foram quantificados como 0,42% &#177; 0,01 em P. alata e 0,10% &#177; 0,01 em P. nitida. No ensaio da atividade antioxidante, a EC50 de P. alata foi de 1061,2 &#177; 8,5 &#181;g/mL e a de nitida 128,0 &#177; 0,9 &#181;g/mL, no ensaio do DPPH, e de 1076 &#177; 85 &#181;&#181;mol de Trolox/g de extrato e de 1985 &#177; 104 &#181;mol de Trolox/g de extrato no ensaio de ORAC, respectivamente. No ensaio de atividade antiúlcera aguda P. alata exibiu, na área total de lesão (ATL), proteção contra as lesões gástricas de 100%, P. nitida de 84% e lansoprazol, o controle positivo, de 76%. Quanto à área relativa de lesão (ARL), alata exibiu proteção contra as lesões de 99,45%; P. nitida, de 82,27% lansoprazol, de 81,44%, no ensaio de lesão gástrica induzida por etanol/HCI com 300 mmol/L de HCI e extratos na dose de 400 mg/kg. As doses de 100,200 e 400 mg/kg dos extratos foram testadas nas mesmas condições com 150 mmol/L de HCI. P. alata apresentou, na ATL, 100% de proteção contra as lesões gástricas nas três concentrações e lansoprazol, de 75%. Na ARL, P. alata exibiu 100% de proteção lansoprazol, de 76,92%. P. nitida apresentou, na ATL, proteção contra as lesões gástricas de 25%, 74% e 94% nas três concentrações, respectivamente e, lansoprazol, de 80%. Na ARL, P. nitida exibiu 27,40%, 74,00% e 91,78% de proteção, respectivamente e lansoprazol, de 78,08%. Baseando-se neste estudo possível distinguir as duas espécies através de características morfoanatômicas das folhas e dos caules e, através do perfil cromatográfico. Ambas as espécies apresentaram atividade antiúlcera promissora. / Passiflora alata Curtis and P. nitida Kunth, Brazilian species, were selected for the comparative pharmacognostic study. P. alata has been used for a long time folk medicine and pharmaceutical preparations. In the pharmacobotanic study, leaves from both species showed similarities in shape, size and in the absence indumenta. They differ by the number of petiolar glands. P. alata presents generally two pairs of glands and P. nitida, one pair. Anatomically, both species showed common characteristics to the Passiflora genera: dorsiventral mesophyll; druses the mesophyll, midrib, cortex, medulla and phloem; collateral vascular bundles. Both species\' midrib presented a biconvex shape, but P. alata\'s was prominently shaped, notably in the abaxial surface. Another differential aspect was observed the stem shape: P. alata was quadrangular and P. nitida was rounded. The tissues sequence presented similarity, with exception to larger collenchyma development in P. alata\'s edges. The phytochemical screening showed presence of flavonoids and absence of alkaloids in both species, in the crude drug and extracts. Formation persistent foam was observed with the crude drug of P. alata. Hemolisis was observed with the crude drug and extract of this species. Flavonoids were quantified as 0.42% &#177; 0.01 in P. alata and 0.10% &#177; 0.01 in P. nitida. In the antioxidant activity assay, the EC50 of P. alata was 1061.2 &#177; 8.5 &#181;g/mL and of P. nitida , 128.0 &#177; 0.9 &#181;g/mL, in the DPPH assay, and 1076 &#177; 85 &#181;mol Trolox/g extract and 1985 &#177; 104 &#181;mol Trolox/g extract in the ORAC assay, respectively. In the antiulcer activity assay P. alata showed, in total lesion area (TLA), protection against gastric lesions 100%, P. nitida 84% and lansoprazole, the positive control, 76%. In relative lesion area (RLA), P. alata showed protection against lesions of 99.45%, P. nitida82.27% and lansoprazole 81.44%, in the HCl/ethanol-induced gastric lesion assay, with HCI 300 mmol/L and extracts at doses of 400 mg/kg. Doses of 100, 200 e 400 mg/kg extracts were tested in the same conditions with HCl 150 mmol/L. P. alata showed, TLA, protection against gastric lesions of 100% in the three concentrations and lansoprazole 75%. In RLA, P. alata showed 100% of protection and lansoprazole 76.92%. P. nitida showed, in TLA, protection against gastric lesions of 25%,74% and 94% in the three concentrations, respectively and lansoprazole 80%. In RLA, nitida showed 27.40%, 74.00% and 91.78% of protection, respectively and lansoprazole 78.08%. Based upon this study it is possible to distinguish both species by leaf and stem morphoanatomic characters and by chromatographic profile. Both species presented promising antiulcer activity.
159

Analyse des [NiFe]-Zentrums und der Kofaktoren im H 2-Sensor von Ralstonia eutropha H16

Gebler, Antje 20 November 2008 (has links)
Zusammenfassung Das beta-Proteobakterium Ralstonia eutropha H16 besitzt zwei [NiFe]-Hydrogenasen, die dem Organismus das Wachstum mit H2 als alleiniger Energiequelle unter aeroben Bedingungen ermöglichen. Reguliert wird die Expression dieser [NiFe]-Hydrogenasen durch ein bakterielles Zweikomponentensystem. Die Signaltransduktionskette besteht aus einer H2-sensierenden, regulatorischen [NiFe]-Hydrogenase (RH), einer Histidin-Proteinkinase und einem Response-Regulator. Um Einblicke in die Struktur-Funktions-Beziehung der H2-sensierenden Komponente zu bekommen, wurden Aminosäure-Austausche in konservierten Bereichen nahe des NiFe-Zentrums der RH durchgeführt. Der Austausch des invarianten Glu13 (E13Q, E13L) resultierte im Verlust der regulatorischen, der H2-oxidierenden und der H/D-Austauschaktivitäten. Spektroskopische Daten wiesen auf ein vollständig assembliertes NiFe-Zentrum hin. Mit Hilfe der ortho-/para-Konversionsaktivität wurde gezeigt, dass dieses Zentrum nach wie vor H2 binden kann. Dies deutet darauf hin, dass eine H2-Bindung am aktiven Zentrum nicht für die regulatorische Funktion der RH ausreicht. Durch den Austausch des Asp15 in His wurde der Konsensus eines konservierten Motivs der Standard-Hydrogenasen hergestellt. Das entstandene RH-Mutantenprotein besaß nur noch eine sehr niedrige H2-oxidierende Aktivität, bei nahezu unveränderter H/D-Austauschrate sowie intakter regulatorischer Aktivität. Dies deutet darauf hin, dass der H2-Umsatz nicht entscheidend für die H2-Sensierung ist. Um Informationen über die Struktur des aktiven Zentrums der RH zu erhalten, war es notwendig, große Mengen RH zu isolieren. Hierfür wurde die Strep-tag-Technologie eingesetzt, die es ermöglichte, die RH als natives Doppeldimer und als homodimeres Protein zu reinigen. Röntgenabsorptionsspektroskopie ergab erstmals, dass sich die Koordination des Nickel-Atoms im aktiven Zentrum sowohl im oxidierten als auch im reduzierten Zustand der RH deutlich von der in Standard-Hydrogenasen unterscheidet. / The beta-proteobacterium Ralstonia eutropha H16 is capable of using H2 as a facultative energy source by means of two distinct, energy-converting [NiFe]-hydrogenases. Transcription of the hydrogenase genes is regulated in response to the availability of H2 via a histidyl-aspartyl phosphorelay comprising a heterodimeric, regulatory [NiFe]-hydrogenase (RH), a histidine protein kinase and a response regulator. In order to gain insights into the mechanism of H2-mediated signal transduction, conserved amino acid residues close to the NiFe active site of the RH were exchanged. Replacement of the strictly conserved Glu13 within the RH large subunit by glutamine and leucine resulted in the loss of the regulatory, H2-oxidizing and hydrogen/deuterium exchange activities. Infrared spectroscopic analysis revealed, that the RH E13Q and E13L derivatives contained a fully assembled NiFe active site and showed para-/ortho-H2 conversion activity. These results indicated that H2-binding at the active site is not sufficient for H2 sensing. Replacement of Asp15, a residue unique in H2 sensors, by histidine restored the consensus of energy-linked [NiFe]-hydrogenases. The resulting RH mutant protein showed only traces of H2-oxidizing activity, whereas the H/D-exchange activity and the regulatory activity were nearly unaffected. H2-dependent signal transduction in the respective mutant strain was less sensitive to O2 than in the wild type. These results suggest that the H2 sensing is independent of H2 turnover. To get insights into the structure of the RH it was necessary to isolate large amounts of RH. By establishing the Strep-tag technology, allowing a fast, mild and efficient purification, it was possible to isolate the native RH-double dimer as well as the modified monodimeric RHstop protein in sufficient amounts for spectroscopic analyses. X-ray absorption spectroscopy showed for the first time that the RH active site undergoes significant structural changes upon exposure to H2.
160

Proteinbiochemische, spektroskopische und röntgenkristallographische Untersuchung der Actinobakteriellen [NiFe]-Hydrogenase aus Ralstonia eutropha

Schäfer, Caspar 05 August 2014 (has links)
Im biogeochemischen Wasserstoffkreislauf erfolgt der überwiegende Teil der H2-Aufnahme aus der Atmosphäre durch die Böden. Erst seit kurzem ist bekannt, dass die Oxidation von Wasserstoff in Böden mutmaßlich durch eine Reihe von Bodenbakterien vermittelt wird, die zur Aufnahme von Wasserstoff in atmosphärischen Konzentrationen befähigt sind. Diese Bakterien codieren [NiFe]-Hydrogenasen einer neuen Gruppe, die als Gruppe 5 der [NiFe]-Hydrogenasen klassifiziert wurde. Auch das beta Proteobakterium Ralstonia eutropha besitzt die Gene einer derartigen Hydrogenase, die aufgrund ihrer Ähnlichkeit zu den sonst überwiegend in Actinobakterien gefundenen Vertretern der Gruppe 5 als „Actinobakterielle Hydrogenase“ (AH) benannt wurde. In der vorliegenden Arbeit wurde die AH aus R. eutropha als erste Gruppe 5-[NiFe]-Hydrogenase in reiner Form isoliert und eingehend durch unterschiedliche biochemische, spektroskopische und röntgenkristallographische Verfahren untersucht. Die hierbei erhaltenen Ergebnisse unterstützen die für Gruppe-5-[NiFe]-Hydrogenasen postulierte Funktion im Erhaltungsstoffwechsel der Organismen unter besonderen Bedingungen, schließen jedoch eine Beteiligung der AH an der hochaffinen Oxidation von Wasserstoff in Böden aus. Jedoch zeigt das Enzym die neuartige Eigenschaft der sauerstoffinsensitiven Wasserstoff-Oxidation, was auf die Anwesenheit eines ungewöhnlichen, durch 1 Aspartat und 3 Cysteine koordinierten [4Fe4S]-Clusters und der vermuteten Kopplung der Elektronentransportketten in der mutmaßlich physiologischen doppeldimeren Form des Enzyms zurückzuführen sein dürfte. Die Arbeit erweitert somit die Kenntnisse auf dem Gebiet der Sauerstofftoleranz von Hydrogenasen sowie der Eigenschaften der Gruppe 5-[NiFe]-Hydrogenasen und ihrer physiologischen Rolle in den betreffenden Organismen. / In the biogeochemical hydrogen cycle, the dominating process for hydrogen uptake from the atmosphere is performed in soils. Only recently it was shown that hydrogen oxidation in soils is presumably mediated by a number of soil-dwelling actinobacteria, which are enabled in high-affinity hydrogen uptake. These bacteria encode [NiFe] hydrogenases of a novel group classified as group 5 of [NiFe] hydrogenases. A hydrogenase of this group is also found in the beta proteobacterium Ralstonia eutropha and was named „Actinobacterial Hydrogenase“ (AH) for its similarity to the group 5 [NiFe] hydrogenases found in actinobacteria. In this work, the AH from R. eutropha was, as the first group 5 [NiFe] hydrogenase, purified to homogeinity and thoroughly characterized by various biochemical, spectroscopic and X-ray crystallographic methods. The results obtained hereby support the function in maintaining a basal metabolism under challenging conditions, that was postulated for group 5 [NiFe] hydrogenases. Yet, the results also exclude the possibility of the AH contributing to high-affinity hydrogen uptake in soils. However, the enzyme shows the novel property of being able of oxygen-insensitive hydrogen oxidation. This property is obviously connected to an unusual [4Fe4S] cluster coordinated by 1 aspartate and 3 cysteines, as well as to a supposed coupling of the electron transport chains in the double dimeric native form of the enzyme. Hence, this work broadens the knowledge in the field of oxygen tolerant hydrogen oxidation and provides new insights in the function of group 5 [NiFe] hydrogenases and their physiological role in the organisms.

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