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Preparation Of Cross-linked Tyrosinase AggregatesAytar, Burcu Selin 01 June 2006 (has links) (PDF)
ABSTRACT
PREPARATION OF CROSS-LINKED TYROSINASE AGGREGATES
Aytar, Burcu Selin
M.S., Department of Chemical Engineering
Supervisor: Prof. Dr. Ufuk Bakir
June 2006, 82 pages
The aim of this study was to prepare cross-linked enzyme aggregate (CLEA) from crude mushroom (Agaricus bisporus) extract. However, the optimization of CLEA production was performed by using pure tyrosinase. Important parameters were determined as protein, ammonium sulfate and glutaraldehyde concentrations, CLEA particle size, and cross-linking temperature and period. On the other hand, the order of ammonium sulfate and glutaraldehyde addition did not affect the yield of CLEA. Optimum CLEA preparation conditions were 60 % ammonium sulfate saturation, 2 % (v/v) glutaraldehyde, and 3 hour cross-linking reaction at room temperature. Particle size of the CLEAs should be reduced by mechanical stirring to eliminate mass transfer limitations. Under these circumstances, 100 % recovery was obtained from both pure and crude tyrosinases. Optimum temperature and the activation energy for catechol oxidation were determined as 34 oC and 16.9 kcal/mol for CLEAs, whereas, 32 oC and 12.5 kcal/mol for the free enzyme. Furthermore, the thermostability of CLEAs was significantly higher than the free enzyme. CLEAs, prepared from crude mushroom extract, retained 72 % of its maximum activity in eight months storage at 4 oC. Moreover, changing the storage temperature from 4 oC to room temperature did not decrease CLEAs stabilities.
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Deletion Mutation Of Glnb And Glnk Genes In Rhodobacter Capsulatus To Enhance Biohydrogen ProductionPekgoz, Gulsah 01 September 2010 (has links) (PDF)
Rhodobacter capsulatus is a photosynthetic, purple non-sulfur (PNS) bacterium that produces biohydrogen via photofermentation. Nitrogenase enzyme is responsible for hydrogen production / during fixation of molecular nitrogen into ammonium, hydrogen is produced. Since this process is an energetically expensive process for the cell, hydrogen production is strictly controlled at different levels. When ammonium is present in the environment, hydrogen production completely ceases. The key proteins in the regulation of nitrogenase by ammonium are two PII proteins / GlnB and GlnK.
&lsquo / Hyvolution&rsquo / , 6th framework EU project, aims to achieve maximum hydrogen production by combining two hydrogen production processes / dark fermentation and photofermentation. In the first stage of the overall process, biomass is used for hydrogen production in dark fermentation process. Then, the effluent of dark fermentation is further utilized by photosynthetic bacteria to produce more hydrogen. However, the effluent of dark fermentation contains high amount of ammonium, which inhibits photofermentative hydrogen production. In order to achieve maximum hydrogen production, ammonium regulation of nitrogenase enzyme in R.capsulatus has to be released. For this purpose, all PII signal transduction proteins of R.capsulatus (GlnB and GlnK) were targeted to be inactivated by site-directed mutagenesis. The internal parts of glnB and glnK genes were deleted individually without using antibiotic cassette insertion. The successful glnB mutant was obtained at the end of mutagenesis studies. In the case of glnK mutation, the suicide vector was constructed and delivered into the cells. However, glnK mutant could not be obtained.
The effect of ammonium on glnB mutant R.capsulatus was investigated and compared with wild type. Biomass of the bacterial cultures, pH of the medium and amount of produced hydrogen were periodically determined. Moreover, the concentrations of acetic, lactic, formic and propionic acids in the medium were periodically measured. Both wild type and glnB mutant grew on acetate and effectively utilized acetate. Ammonium negatively affected hydrogen production of glnB mutant and wild type. The ammonium inhibition of hydrogen production did not release in glnB mutant due to the presence of active GlnK protein in the cell / hence, inactivation of one of PII proteins was not enough to disrupt ammonium regulation of the cell. Moreover, kinetic analysis of bacterial growth and hydrogen production were done. Growth data fitted to the Logistic Model and hydrogen production data fitted to the Modified Gompertz Model.
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Ammonium And Lead Exchange In Clinoptilolite Zeolite ColumnBahaalddin, Ahmad Dh. 01 January 2011 (has links) (PDF)
Wastewaters resulted from anthropogenic influence can encompass a wide range of potential contaminants and concentrations. There are numerous procedures that can be used to clear out wastewaters depending on the type and extent of contamination, however / disposal of pollutants from wastewaters in industrial scale is a difficult and costly problem.
In this study, the use of ion exchange theory utilizing natural Turkish clinoptilolite zeolite from Gö / rdes-Manisa as ion exchange resins in down-flow column mode is investigated. The clinoptilolite with particle size range of 0.25-0.50 mm is used in the removal of lead Pb2+ and ammonium NH4+ ions from aqueous solutions.
The aim of the study is to set up the conditions under which clinoptilolite may be used in an economical and efficient approach in the removal process. Experiments were divided into two sets: binary studies, and ternary studies, and the effects of conditioning clinoptilolite with NaCl solution, flow rate, and initial concentration of the solutions on the removal behavior were investigated.
In binary studies, results showed that increasing the loading volumetric flow rate resulted in decreasing the breakthrough capacity and the column efficiency, while the total capacity remained constant. The maximum total capacity was determined as 1.16 meq/g of zeolite for NH4+, and 1.1 meq/g of zeolite for Pb2+ and these values were close to each other and to the sodium content of Na-form of pretreated clinoptilolite (1.16 meq/g of zeolite). In addition, by decreasing the initial contaminant concentration, an increase in breakthrough capacity and column efficiency was observed.
In ternary studies, the results showed that the removal of Pb2+ and NH4+ ions are dependent on the flow rate, in which at moderately low flow rate, a higher ion exchange capacity is yielded. That was explained as at higher flow rates, the retention time was insufficient for the ion exchange process to take place completely between clinoptilolite and lead and ammonium ions. Thus, a competition between Pb2+ and NH4+ ions for the exchange sites on clinoptilolite was observed and this competition was in favor of lead ions.
Consequently, it was observed that the clinoptilolite zeolite has affinity for both Pb2+ and NH4+ ions. However, the affinity of clinoptilolite for lead ions is higher than that for ammonium ions. Therefore, the cations selectivity for clinoptilolite according to their affinity is determined as the following sequence: NH4+ > / Pb2+ > / Na+.
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Study of Mechanisms of Secondarily Treated Sewage and Textile Wastewater by Hybrid Constructed WetlandsChuang, Hsiao-hui 13 February 2009 (has links)
The aim of this investigation was to use hybrid constructed wetlands to treat the secondary effluents from NSYSU campus sewage treatment plant, which had high phosphate and ammonium nitogen and from a textile industrial wastewater treatment plant, which had high chemical oxygen demand(COD) . The purpose of this study is to design optimum operation, conditions and to select suitable types of filter media through optimum combinations of vertical flow (VF) and horizontal flow (HF) constructed wetland systems.
The flow regimes for vertical flow operation in this study include continuous flow with filled water, trickling filter type and batch type, while the flow types for horizontal flow operation include high water level and low water level effluents. The experimental of results showed that the best ammonium nitrogen removal efficiency was found in trickling filter type, which was because high oxygen was provided under this flow pattern creating a suitable condition for nitrification , especially in V3 column(39.09%), while the best denitrification effect was fonnd in low water level horizontal operation, especially in H2 bed(42.56%).
The experimental results of treating the Everest effluent from the wastewater treatment plant showed that the flow regime in V3 system had best removal of COD in batch type. In trickling filter and low water level type, the optimum hybrid of V3+H3 had the COD removal efficiency eqail to(33.3%)+(49.8%) respectirely .For the experimental results of tolerance of macrophyte, Hedycbium coronarium Koenig live well, but no significant removal efficiencies of nutrient was fund.
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Starch crosslinking for cellulose fiber modification and starch nanoparticle formationSong, Delong 23 March 2011 (has links)
As a low cost natural polymer, starch is widely used in paper, food, adhesive, and many other industries. In order to improve the performance of starch, crosslinking is often conducted either in the processes of starch modification or during the application processes. Many crosslinkers have been developed in the past for crosslinking starch. Ammonium zirconium carbonate (AZC) is one of the common crosslinkers for crosslinking starch in aqueous solutions, having been widely used as a starch crosslinking agent in paper surface coating for more than 20 years. However, the mechanisms of starch crosslinking with AZC have not been well studied. In order to optimize the crosslinking chemistry of starch and find new paths for the utilization of starch in papermaking, a better understanding of the starch crosslinking mechanism is necessary.
This thesis focuses on the fundamental study of starch crosslinking in an aqueous solution and its applications in fiber surface grafting, filler modification, and starch nanoparticle formation. Particularly, the thesis contains three major parts:
(1) Mechanism study of starch crosslinking induced by AZC:
In this thesis, the crosslinking (or gelation) kinetics of starch/AZC blends were investigated by using rheological measurements. The evolution of viscoelastic properties of AZC solutions and the AZC-starch blends was characterized. It was found that for both AZC self-crosslinking and AZC-starch co-crosslinking, the initial bond formation rate and the gel strength had a strong power law relationship with the concentrations of both AZC and starch. It is suggested that the development of the crosslinking network is highly dependent on the AZC concentration, while the starch concentration effect is less significant. It was determined that the activation energy of AZC self-crosslinking was approximately 145-151 kJ/mol, while the activation energy of AZC-starch co-crosslinking was 139 kJ/mol.
(2) Fiber and filler modifications with starch and crosslinkers:
Besides reacting with starch, AZC can react with cellulose which also contains hydroxyl groups. Theoretically, it is possible to use AZC as a crosslinker / coupling agent to graft starch onto cellulose fibers. It is believed that the grafted starch on fiber surfaces can improve the fiber bonding capability. In this thesis, a facile method to graft starch onto cellulose fiber surfaces through the hydrogen bond formation among cellulose, starch and AZC was developed. Compared with the paper sheets made of fibers with an industry refining level (420 ml CSF), the paper sheets made of fibers with a much lower refining degree but with grafted starch showed higher paper strengths, including the tensile strength, stiffness and z direction tensile; meanwhile, a faster drainage rate during web formation could also be achieved.
Not only can the fiber-fiber bonding be improved by grafting starch onto fiber surfaces, but the filler-fiber bonding can also be improved if starch can be effectively coated on the filler surface. This concept has been supported by the early studies. In this thesis, the effects of the crosslinking of starch in the filler modification for the papermaking application were also studied.
(3) Mechanism of starch nanoparticle formation during extrusion with crosslinkers:
It was reported that starch crosslinking could facilitate the reduction of starch particle size during reactive extrusion. However, the mechanism of the particle size reduction by starch crosslinking was not illustrated. The reason that the crosslinking can cause the particle size reduction of starch during extrusion is fundamentally interesting. In this thesis, the mechanism of starch particle size reduction during extrusion with and without crosslinkers was investigated by identifying the contributions of thermal and mechanical effects. The effects of extrusion conditions, including temperature, screw speed, torque, starch water content and crosslinker addition, on the particle size were studied. It was found that the addition of crosslinkers could significantly increase the shear force (torque), and consequently facilitate the reduction of the particle size. The results indicate that for extrusion without a crosslinker, the starch particle size decreased with the increase of temperature. At 100 degree Celsius, the starch particles with a size of 300 nm could be obtained. With the addition of appropriate crosslinkers (glyoxal), the starch particle size could be reduced to around 160 nm, even at a lower extrusion temperature of 75 degree Celsius .
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Fate and effect of quaternary ammonium antimicrobial compounds on biological nitrogen removal within high-strength wastewater treatment systemsHajaya, Malek Ghaleb 20 May 2011 (has links)
High strength wastewater (HSWW) generated in food processing industries is characterized by high organic carbon and nitrogen content, and thus high oxygen demand. Biological nitrogen removal (BNR) is a technology widely used for the treatment of HSWW. Food processing facilities practice sanitation to keep food contact surfaces clean and pathogen-free. Benzalkonium chlorides (BACs) are cationic quaternary ammonium antimicrobial compounds (QACs) common in industrial antimicrobial formulations. BAC-bearing wastewater generated during sanitation applications in food processing facilities is combined with other wastewater streams and typically treated in BNR systems. The poor selectivity and target specificity of the antimicrobial BACs negatively impact the performance of BNR systems due to the susceptibility of BNR microbial populations to BAC. Objectives of the research were: a) assessment and quantification of the inhibitory effect of QACs on the microbial groups, which mediate BNR in HSWW treatment systems while treating QAC-bearing HSWW; b) evaluation of the degree and extent of the contribution of QAC adsorption, inhibition, and biotransformation on the fate and effect of QACs in BNR systems. A laboratory-scale, multi-stage BNR system was continuously fed with real poultry processing wastewater amended with a mixture of three benzalkonium chlorides. The nitrogen removal efficiency initially deteriorated at a BAC feed concentration of 5 mg/L due to complete inhibition of nitrification. However, the system recovered after 27 days of operation achieving high nitrogen removal efficiency, even after the feed BAC concentration was stepwise increased up to120 mg/L. Batch assays performed using the mixed liquors of the BNR system reactors, before, during, and post BAC exposure, showed that the development of BAC biotransformation capacity and the acquisition of resistance to BAC contributed to the recovery of nitrification and nitrogen removal. Kinetic analysis based on sub-models representing BNR processes showed that BAC inhibition of denitrification and nitrification is correlated with BAC liquid-phase and solid-phase concentrations, respectively. Simulations using a comprehensive mathematical BNR model developed for this research showed that BAC degradation and the level of nitrification inhibition by BAC were dynamic brought about by acclimation and enrichment of the heterotrophic and nitrifying microbial populations, respectively. The fate and effect of BACs in the BNR system were accurately described when the interactions between adsorption, inhibition, and resistance/biotransformation were considered within the conditions prevailing in each reactor. This work is the first study on the fate and effect of antimicrobial QACs in a continuous-flow, multi-stage BNR system, and the first study to quantify and report parameter values related to BAC inhibition of nitrification and denitrification. Results of this study enable the rational design and operation of BNR systems for the efficient treatment of QAC-bearing wastewater. The outcome of this research provides information presently lacking, supporting the continuous use of QACs as antimicrobial agents in food processing facilities, when and where needed, while avoiding any negative impacts on biological treatment systems and the environment.
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Effekten på nedbrytningen av rötter vid tillförsel av ammonium sulfat i en granskog i sydvästra Sverige / Effect of ammonium sulphate addition on root decomposition in a Norway spruce stand in south-west SwedenGustafsson, Therése January 2002 (has links)
<p>Decomposition of organic matter is a critical process in the ecosystem, which involves many essential biotic and physical parts. Decomposition is therefore an important process both above and below ground. The rate of decomposition is dependent of many environmental factors for example: pH, moisture and supply of oxygen. The decomposition can therefore be affected by large scaled environmental influences, such as acidification and climatic changes. The root litter in the forest is in different ways affected by acidification, liming and manuering. Because of the important role the root system has to the whole forest ecosystem, it can be of importance to gain knowledge about how roots are affected by external environmental influences. </p><p>In the forest ecosystem fertilise the soil has become a common practice in forest management to optimise tree production. Experiments with nitrogen fertilisation have shown that the volume growths of the tree and litter supply have increased after fertilisation. There are also reports about the negative effects nitrogen fertilisation has on decomposition, which results in a decreased decomposition of organic matter. </p><p>The aim of this study is to investigate how the decomposition of organic matter, in this case roots, is affected by a large addition of ammonium sulphate. The study concentrates on to statistically evaluate important aspects on how addition of ammonium sulphate affects the decomposition of organic matter below ground in different soil layers and root diameters, and investigate the possibilities that addition of ammonium sulphate could lead to a decreasing potential of carbon mineralisation. </p><p>The study was conducted is in Skogaby, which is located in southwest Sweden in the community of Halland. Samplings of roots were made in the experimental area from the humus and mineral layer. Roots used for this study varied from less than 2mm up to 2-5mm. Decomposition of root litters were made with litterbags, which were placed in the soil in the humus and mineral layer in the original place of were the roots were collected. The results from this study showed that there appear significant differences in some of the cases between the control and ammonium sulphate treatments. The conclusion that can be drawn by this study is that the addition of ammonium sulphate, under certain conditions depending on root diameter and soil layer, comes to affect the decomposition of root litter. The addition of ammonium sulphate seems to have a positive effect on the decomposition in the initial phase, for then come to decrease in the later phases and be similar to the control areas. It can also be determined that decomposition does not seem to vary within treatments with regards to root diameter and soil layer. Regarding the question about how carbon mineralisation is affected by addition of ammonium sulphate it is probable that the addition would come to increase the mineralisation in the initial phases of the decomposition, compared with the control plots.</p>
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Imagerie spectroscopique et spectroscopie localisée par RMN du proton in vivo.<br />Application à l'étude de la physiopathologie tumorale par cartographie des métabolites et du pH extracellulaire dans un modèle experimental de gliomes C6Provent, Peggy 13 December 2006 (has links) (PDF)
L'objectif de ce travail était d'optimiser les conditions ainsi que les techniques d'acquisition par spectroscopie et par imagerie spectroscopique par RMN du proton in vivo, chez le petit animal pour les appliquer à des études biologiques. Les conditions d'acquisitions ont été améliorées par la mise en place d'une technique de correction automatique des inhomogénéités de champ magnétique. Les séquences d'acquisition ont été optimisées par l'utilisation de la technique de suppression d'eau VAPOR et par l'utilisation de l'imagerie spectroscopique par encodage spiral à temps d'écho court. <br />Le pH extracellulaire (pHe) dans les tumeurs est plus acide que dans le tissu sain. Cette acidité semble être un facteur favorisant la prolifération. A l'aide d'une nouvelle molécule sonde, l'ISUCA, nous avons utilisé l'imagerie spectroscopique pour cartographier la distribution de pH extracellulaire dans les tumeurs et la comparer avec la distribution du lactate. Celles-ci ne sont pas corrélées spatialement, de plus à la suite d'une hyperglycémie, les quantités de lactate et de protons augmentent de manière globale mais avec une corrélation spatiale négative. Ces résultats suggèrent une redistribution des protons des sites de la glycolyse vers des pompes membranaires.<br />Le second projet biologique portait sur l'étude du rôle de l'ammonium dans la régulation de la glycolyse dans le cerveau. L'hyperammoniémie augmente la concentration de lactate cérébral: j'ai montré que cette augmentation peut être transitoire, ce qui est une condition incontournable pour un éventuel rôle physiologique de l'ammonium dans le couplage métabolique entre neurones et astrocytes.
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Reactive replacement and addition of cations in bioclastic silica and calciteAllan, Shawn Michael. January 2005 (has links)
Thesis (M. S.)--Materials Science and Engineering, Georgia Institute of Technology, 2005. / Committee Chair: Kenneth Sandhage; Committee Members: Joe Cochran, Robert Snyder and Tom Sanders. Part of the SMARTech Electronic Thesis and Dissertation Collection.
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Investigation of the Optimal Dissolved CO2 Concentration and pH Combination for the Growth of Nitrifying BacteriaMorris, Raymond Anthony 01 January 2011 (has links)
Ammonium (NH4+) is a biological nutrient that is transformed in a wastewater treatment plant (WWTP) in a process called activated sludge. This is accomplished in an aerobic environment using microorganisms and inorganic carbon that convert the ammonium to nitrate (NO3-). This process is termed nitrification. Removal of ammonium is necessary due to its oxygen demand and toxicity to the environment.
Nitrification is considered a slow process due to the slow growth rate of the nitrifying bacteria. Ammonia oxidizing bacteria (AOB) first covert the ammonium (NH4+) to nitrite (NO2-) followed by conversion to nitrate (NO3-) by nitrite oxidizing bacteria (NOB). These slow rates limit the treatment capacity of the WWTP.
The initial hypothesis suggested that these slow rates were due to limited carbon in the aeration basin of a WWTP. A series of designed experiments and observational studies revealed substantial dissolved CO2 exists throughout a WWTP. Based on these findings, the central research focused on determining if an optimum dissolved CO2 concentration/ pH combination exists that maximizes nitrification.
Experimentation conducted at a pH of 7.0 and varying concentrations of dissolved CO2 concentration revealed inhibition at low (<5 mg/l) and high (>30 mg/l) dissolved CO2 concentration levels. Further research found that optimum nitrification can be attained in a dissolved CO2 concentration range of 10 - 15 mg/l and a pH range of 7.5 - 8.0. A maximum specific growth rate of 1.05 - 1.15 days-1 was achieved. A partitioning of the sums of squares from these designed experiments found that pH accounts for approximately 83 percent of the sums of squares due to treatment with the dissolved CO2 concentration accounting for 17 percent. This suggests that pH is the dominant factor affecting nitrification when dissolved CO2 concentration is optimized.
Analysis of the growth kinetics for two of the designed experiments was conducted. However, a set of parameters could not be found that described growth conditions for all operating conditions. Evaluating the results from these two experiments may suggest that a microbial population shift occurred between 16 and 19 mg/l of dissolved CO2 concentration. These dissolved CO2 concentrations represent pH values of 7.1 and 7.0, respectively, and were compared to experimentation conducted at a pH of 7.0. Though the pH difference is minor, in combination with the elevated dissolved CO2 concentration, a microbial shift was hypothesized.
Microbial samples were collected from the designed experiment that optimized dissolved CO2 concentration (5, 10 and 15 mg/l) and pH (6.5, 7.0, 7.5 and 8.0). These samples were evaluated using Fluorescence in situ hybridizations (FISH) to determine the population density of common ammonium oxidizing bacteria (AOB) (Nitrosomonas and Nitrosospira) and nitrite oxidizing bacteria (NOB) Nitrobacter and Nitrospirae). The dominant AOB and NOB microbes were found to be Nitrosomonas and Nitrospirae.
These results suggest that increased nitrification rates can be achieved by incorporating appropriate controls in a wastewater treatment plant (WWTP). With higher nitrification rates, lower nitrogen values can be obtained which will reduce the WWTP effluent nitrogen concentration. Conversely, these increased nitrification rates can also reduce the volume of an aeration basin given similar effluent nitrogen concentrations.
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