In situ recovery of secondary metabolites using adsorption resins : a thesis presented in partial fulfilment of the requirements for the degree of Master of Philosophy at Massey University, Palmerston North, New Zealand

Almost without exception a two to three fold increase in microbial secondary metabolite concentration was measured when adsorption resins were added in-situ during a submerged liquid fermentation. Anguidine was produced at a final concentration of 440 mg/L after five days in a shake flask that contained adsorption resin, compared to 300 mg/L without resin. Rapamcyin was produced at a final concentration of 87 mg/L after six days in a shake flask that had resin present, compared to 28 mg/L without resin. Ansamitocin P3 was produced at a final concentration of 24 mg/L after six days in a shake flask with resin, compared to 9.75 mg/L without resin. The increase in secondary metabolite concentration confirmed that the resins used provided a positive influence on secondary metabolite production. Adsorption resins for shake flask studies were selected based on their ability to achieve maximum adsorption of specific secondary metabolites in various fermentation systems. A library of adsorbed concentrations was collected for the three secondary metabolites studied. The lipophilicty of the metabolite, calculated by several software packages, was compared to the polarity of the adsorption resin to generate a relationship. By using the preceding set of data it is possible to select adsorption resins that improved the produced concentrations of the target organic secondary metabolites. The fermentation media compositions tested appeared to have no effect on the final product concentration when adsorption resins were added in situ during the fermentations. Based on the lipohilictiy of the secondary metabolite and the polarity of the resins, it is possible to select a resin that achieves a high adsorption concentration of the target organic secondary metabolite

On the Complexity of Random Quantum Computations and the Jones Polynomial

There is a natural relationship between Jones polynomials and quantum computation. We use this relationship to show that the complexity of evaluating relative-error approximations of Jones polynomials can be used to bound the classical complexity of approximately simulating random quantum computations. We prove that random quantum computations cannot be classically simulated up to a constant total variation distance, under the assumption that (1) the Polynomial Hierarchy does not collapse and (2) the average-case complexity of relative-error approximations of the Jones polynomial matches the worst-case complexity over a constant fraction of random links. Our results provide a straightforward relationship between the approximation of Jones polynomials and the complexity of random quantum computations.Comment: 8 pages, 4 figure

Spectroelectrochemical Elucidation of the Kinetics of Two Closely Spaced Electron Transfers

The use of spectroelectrochemistry to facilitate the analysis of an EE mechanism was reported in this work. Using a set of spectra as a function of potential, the spectra of all three oxidation states were determined using evolving window factor analysis. From these spectra, the concentration of each species in solution was determined for each potential. Using these data, the current was calculated. Unlike the direct measurement of current, the current due to each redox process was determined, allowing one to analyze each redox process separate from the other. With the use of the Butler–Volmer equation, the redox potential and the heterogeneous electron transfer parameters were measured. The spectrally determined current has the advantage of determining the current due to each redox process which is not generally possible with voltammetric data when the redox potentials are close together. This method was applied to the spectroelectrochemical reduction of Escherichia coli sulfite reductase hemoprotein (SiR-HP) in a phosphate buffer and in the presence of cyanide. The electrochemical parameters (E°’s, k°’s and α’s) for each electron transfer were calculated for both the uncoordinated and cyanide coordinated species. The rates of electron transfer for the siroheme and iron–sulfur cluster were slower than the rates observed for other heme proteins. This is probably due to the fact that this protein is significantly larger than most of the heme protein previously studied. This approach is a powerful tool for two-electron transfers when the E° values are close together

Molecular and Biological Characterization of a Cryptosporidium molnari-Like Isolate from a Guppy (Poecilia reticulata)

Histological, morphological, genetic, and phylogenetic analyses of a Cryptosporidium molnari-like isolate from a guppy (Poecilia reticulata) identified stages consistent with those of C. molnari and revealed that C. molnari is genetically very distinct from all other species of Cryptosporidium. This study represents the first genetic characterization of C. molnari

Use of Evolutionary Factor Analysis in the Spectroelectrochemistry of Escherichia coli Sulfite Reductase Hemoprotein and a Mo/Fe/S Cluster

The deconvolution of spectroelectrochemical data is often quite difficult if the spectra of intermediates are not known. Factor analysis, however, has been shown to be a powerful technique which can make it possible to deconvolute overlapping spectra. In this work, evolving factor analysis will be used to determine the number of intermediates and the spectra of those species for two typical spectroelectrochemical experiments:  linear scan voltammetry and chronoabsorptometry in a thin-layer cell. The first system was the reduction of E. coli sulfite reductase hemoprotein (SiR-HP). Principal factor analysis indicated that three species were present. By using evolving factor analysis, the potential regions where each of the species were present were identified, and their concentrations and spectra were determined by the use of the mass balance equation. The spectra of the one-electron (SiR-HP1-) and two-electron (SiR-HP2-) reduced product were compared with previous work. The second experiment was the chronoabsorptometry of Cl2FeS2MoS2FeCl22- in methylene chloride. This experiment indicated that five species were present during the experiment. The entire set of 61 spectra were fit by assuming that there were 4 species present during the electrolysis. The rate constant for the appearance of subsequent species fit quite well with the rate constant for the disappearance of previous species. The spectra of the intermediates and final product were obtained using evolving factor analysis and a mass balance equation. Identification of the fifth species, which was probably the initial reduction product, Cl2FeS2MoS2FeCl23-, was difficult due to its low concentration and the fact that it was present in the same time region as the starting material