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Red yeast epoxide hydrolases : growth, activity and selectivity / J. MaritzMaritz, Jana January 2007 (has links)
Enantiopure epoxides are versatile compounds in the production of single enantiomer drugs, and are of high value as building blocks and intermediates in the preparation of more complex single enantiomer pharmaceuticals and agrochemicals.
Epoxide hydrolases, ubiquitous enzymes in nature, can be versatile tools in the biocatalytic production of these single enantiomer epoxides due to their capability of selectively hydrolysing one enantiomer of a wide range of these compounds, and thus rendering an enantiopure epoxide and diol. The value of epoxide hydrolases for the kinetic resolution of epoxide compounds are dependant on factors such as availability, ease of production, long term stability, activity and the displayed enantioselectivity.
The first objective of this study was to investigate and optimise the growth media and time for the production of two red yeasts, Rhodotorula glutinis and Rhodospondium toruloides, and their epoxide hydrolysing enzymes. Maximum and minimum epoxide hydrolase (EH) activity for R. glutinis was respectively observed with the YMvit (0,26 mM.min"1) and malt (0,17 mM.min"1) media, while peak biomass production was observed from the YM medium (64,9 mg.mL"1). For R. toruloides, the highest biomass was produced in the YM (130,8 mg.mL"1) medium, with similar epoxide hydrolase activities (average c = 0,75 ± 0,01) displayed for the YM, YMvit and malt grown biocatalysts.
With varying the YM medium glucose concentration (0,5 - 2,0 %) the most biomass was produced for R. glutinis with the addition of 1,5 % glucose (60,0 + 0,9 mg.mL"1), with a slight drop in the biomass observed with the addition of 2,0% glucose (56,0 + 1,7 mg.mL"1). No significant differences in epoxide hydrolase activity was observed for the lower glucose additive concentrations (0,5 - 1,5 %), while 2,0 % (m/v) rendered a biocatalyst with almost 20 % higher activity (0,29 mM.min"1). For R. toruloides an increase in the glucose concentration lead to a significantly higher biomass production while the time needed to attain the stationary phase increased progressively from 40 to 96 hours. Almost equal activity was observed for the top three glucose concentrations (average c = 0,82 ± 0,01) at 36 hours growth time, but in all cases a decrease in the EH activity was observed during the stationary phase, with the most pronounced decrease for the 2,0 % (m/v) glucose concentration, that showed a drop in conversion of almost 62 % at 144 hours growth time.
The second objective was to synthesise meta and para nitro-, methyl- and methoxystyrene oxides and the successive production of their single enantiopure epoxides through R. glutinis
EH mediated kinetic resolution, and the determination of the absolute configuration of the pure residual enantiomers through VCD analysis. R. glutinis selectively hydrolysed the whole range of styrene oxide derivatives, with the highest activity displayed towards the meta substituted derivatives in the order of methyl > methoxy > nitro. m-Methylstyrene oxide reached a % e.e. of >98 within 60 minutes, with an exceptionally high yield of 42,5 %. The absolute configuration of the residual epoxide enantiomers of /n-nitro, m-methyl and m-methoxystyrene oxides were determined to be of the (S)-configuration, indicating that R. glutinis EH preferentially hydrolyses the (R)-epoxides.
Thirdly, we attempted to increase the R. glutinis EH activity through the addition of hydroxypropyl-p-cyclodextrin (HPB) and to correlate the rate of chemical and R. glutinis EH mediated enzymatic hydrolysis, and the enzyme's enantioselectivity to the electronic properties of their substituents and the spatial arrangement of the substrates in relation to the EH catalytic triad of the EH active site.
An increase in the HPB concentration (0 - 20 % w/v) lead to a substantial increase in both the solubility as well as enzyme activity for p-N02 (para-nitrostyrene oxide) with a significant increase in the solubility of between 2,89 and 6,28 times for the substrate range with the addition of 5 % HPB in comparison to the buffer solution.
The acid induced chemical and R. glutinis EH mediated enzymatic reaction rate was correlated to both the Hammett constant as well as the Mulliken charge distributions. The Mulliken charge distribution over the protonated epoxides was correlated to the acid induced chemical hydrolysis rates, while the Mulliken charge distribution over the neutral epoxides could be correlated to the enzymatic reaction rates. An increase in the electron-donating properties of the styrene oxide substituent groups was correlated to an increase in both the chemical as well as the R. glutinis EH mediated hydrolysis reaction rates of the styrene oxide derivatives.
Docking of the possible conformers of the (R)- and (S)-enantiomers of these meta and para substituted styrene oxides into the EH binding site of the closely related Aspergillus niger displayed a closer and more preferential fit of the (R)-epoxides which is the faster reacting enantiomerfor both A. niger and R. glutinis EHs.
The proven relationship between R. glutinis EH activity and selectivity and the electronic properties of substituent groups, as well as the relationship between spatial arrangement of the epoxide hydrolase binding site and the enantioselectivity of the enzyme, could open up the possibility to correctly predict both the enantioselectivity as well as the activity of R. glutinis EH, and possibly other red yeasts, towards more complex epoxide substrates without the need of time consuming screenings. / Thesis (Ph.D. (Pharmaceutical Chemistry))--North-West University, Potchefstroom Campus, 2008.
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Problems for a Kantian account of disputes over judgments of tasteMeadows, Toby Scougall, History & Philosophy, Faculty of Arts & Social Sciences, UNSW January 2007 (has links)
In this thesis, we will be concerned with Kant's philosophy of beauty and with a particular area of interest which is of great importance to any practical theory of aesthetics: dispute. If one judges something to be beautiful - if one makes a judgment of taste, and we seek to persuade others to our conviction, then how can we go about doing this? We shall discover that Kant's theory provides an interesting account of how disputes over taste can take place: an account which is informative for the way in which we conduct aesthetics today.
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Multi-resolution indexing method for time seriesMa, Mei January 2010 (has links)
Time series datasets are useful in a wide range of diverse real world applications. Retrieving or querying from a collection of time series is a fundamental task, with a key example being the similarity query. A similarity query returns all time series from the collection that are similar to a given reference time series. This type of query is particularly useful in prediction and forecasting applications. / A key challenge for similarity queries is efficiency and for large datasets, it is important to develop efficient indexing techniques. Existing approaches in this area are mainly based on the Generic Multimedia Indexing Method (GEMINI), which is a framework that uses spatial indexes such as the R-tree to index reduced time series. For processing a similarity query, the index is first used to prune candidate time series using a lower bounding distance. Then, all remaining time series are compared using the original similarity measure, to derive the query result. Performance within this framework depends on the tightness of the lower bounding distance with respect to the similarity measure. Indeed much work has been focused on representation and dimensionality reduction, in order to provide a tighter lower bounding distance. / Existing work, however, has not used employed dimensionality reduction in a flexible way, requiring all time series to be reduced to have the same dimension. In contrast, in this thesis, we investigate the possibility of allowing a variable dimension reduction. To this end, we develop a new and more flexible tree based indexing structure called the Multi-Resolution Index (MR-Index), which allows dimensionality to vary across different levels of the tree. We provide efficient algorithms for querying, building and maintaining this structure. Through an experimental analysis, we show that the MR-Index can deliver improved query efficiency compared to the traditional R-tree index, using both the Euclidean and dynamic time warping similarity measures.
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Multi-resolution indexing method for time seriesMa, Mei January 2010 (has links)
Time series datasets are useful in a wide range of diverse real world applications. Retrieving or querying from a collection of time series is a fundamental task, with a key example being the similarity query. A similarity query returns all time series from the collection that are similar to a given reference time series. This type of query is particularly useful in prediction and forecasting applications. / A key challenge for similarity queries is efficiency and for large datasets, it is important to develop efficient indexing techniques. Existing approaches in this area are mainly based on the Generic Multimedia Indexing Method (GEMINI), which is a framework that uses spatial indexes such as the R-tree to index reduced time series. For processing a similarity query, the index is first used to prune candidate time series using a lower bounding distance. Then, all remaining time series are compared using the original similarity measure, to derive the query result. Performance within this framework depends on the tightness of the lower bounding distance with respect to the similarity measure. Indeed much work has been focused on representation and dimensionality reduction, in order to provide a tighter lower bounding distance. / Existing work, however, has not used employed dimensionality reduction in a flexible way, requiring all time series to be reduced to have the same dimension. In contrast, in this thesis, we investigate the possibility of allowing a variable dimension reduction. To this end, we develop a new and more flexible tree based indexing structure called the Multi-Resolution Index (MR-Index), which allows dimensionality to vary across different levels of the tree. We provide efficient algorithms for querying, building and maintaining this structure. Through an experimental analysis, we show that the MR-Index can deliver improved query efficiency compared to the traditional R-tree index, using both the Euclidean and dynamic time warping similarity measures.
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Court-connected alternative dispute resolution : the Ontario mandatory mediation program and the German obligatory settlement procedure. Approaches to achieving early case settlement: a comparative analysis.Gilbert, Martin. January 2004 (has links)
Thesis (LL. M.)--University of Toronto, 2004. / Adviser: Alan Stitt.
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The adoption of dispute resolution advisor system for preventing and resolving dispute in Hong Kong construction industryLeung, Nga Yee Ruth. January 2008 (has links) (PDF)
Thesis (M.A.)--City University of Hong Kong, 2008. / "Master of Arts in arbitration and dispute resolution." Title from PDF t.p. (viewed on June 1, 2009) Includes bibliographical references.
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Super resolution of fingerprintsDeshpande, Kaustubh R. January 1900 (has links)
Thesis (M.S.)--West Virginia University, 2004. / Title from document title page. Document formatted into pages; contains vii, 104 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 95-104).
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Sensitive absorption-based laser wave-mixing spectroscopic methods in microsystems for biomedical applications /Lopez, Mirna Maritza, January 2004 (has links)
Thesis (Ph. D.)--University of California, San Diego, and San Diego State University, 2004. / Vita. Includes bibliographical references (leaves 215-217).
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Dispute resolution under tax treaties /Altman, Zvi Daniel. January 1900 (has links)
Thesis (doctoral) - Harvard Law School, Cambridge, Mass., 2005. / Includes bibliographical references.
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ADR in the age of contemporaneity : complexity, chaos and pedagogy /Martins, Nadia Bevilaqua. January 2003 (has links) (PDF)
Thesis (Ph.D.) - University of Queensland, 2004. / Includes bibliography.
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