Faculty of Applied Sciences
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Item Synthesis, spectroscopic properties and cytotoxicity of boron- dipyrromethene fluorescent dyes(2015-01-14) Bipath, Nirvashini; Gengan, Robert Moonsamy; Chuturgoon, Anil A.In this study, we report the synthesis of three quinolone bearing imidazole derivatives 2, 3 and 4 and two quinolone bearing BODIPY dyes 5 and 7. In the synthesis of 2, 3 and 4, the first step was the preparation of the starting compound 2-chloro-3-formyl quinoline (1); the Vilsmeier-Haack cyclisation protocol was used. Compound 1 was used with the appropriate diamine, together with POCl3 to produce 2, 3 and 4. These compounds were characterized by IR, 1H-NMR and 13C-NMR. In the synthesis of 5, compound 1 was used whilst 6 was used for the synthesis of 7. This was via. a one-pot synthesis using conventional reflux apparatus and Schlenk technique. These compounds were characterized by IR, 1H-NMR and 13C-NMR. Four other BODIPY dyes were also synthesized but their purification by column chromatography were unsuccessful. However a HPLC method was developed using 2 as a model; the best eluting solvent was 65 % methanol. After synthesis, 2, 3, 4, 5 and 7 were used for spectroscopic studies by UV-visible and fluorescence spectroscopy. In the UV-visible studies, 2, 3 and 4 were dissolved, separately, in five solvent viz. ethanol, methanol, dichloromethane, chloroform and acetonitrile. The UV profile of each compound was obtained and the maximum absorbance was then used for fluorescence studies. In the fluorescence studies, all the compounds displayed a fluorescence nature when excited with the various wavelengths. The fluorescence properties, namely Stoke shift, quantum yield, life time, molar absorptivity and brightness, were investigated to establish the properties of each compound in all five solvent systems. The Stoke shift was evident in all compounds and the quantum yields were below one which indicates no other electron transfer mechanisms occurring. The results displayed a favorable response and this further lead to analysis of the synthesized compounds for it potential application as a chemosensor. Eight metal ions were used to investigate this property. All eight metal ions, when reacted with the synthesized compounds, as ligands, showed chemosensor properties, viz. photon induced electron transfer, inter-molecular charge transfer and fluorescence resonance electron transfer, as a quenching and enhancement of emission and excitation peaks were observed. The compounds were further investigated for its potential for its use as a photovoltaic cells. The energies of the compounds were obtained from the analyses of the reflectance and transmission spectra. It was found that the synthesized compounds displayed properties which were positive for its use as a photovoltaic cell. Biological analyses using molecular docking analyses and MTT assays were conducted to determine the use of these as an anti-cancer drug. Compounds 2 and 3 formed hydrogen bonds with GLU 25 and LEU 27, respectively with MDM2-p53 proteins. Following the molecular docking studies, the MTT assay was performed on all five synthesized compounds. The BODIPYs with the quinoline moieties demonstrated a reduction in the rate of A549 cell proliferation when compared to the imidazole and benzimidazoles; this was observed for compounds 5 and 7. Further, a comparison between imidazoles clearly shows that compounds 3 and 4 also decreased cell proliferation. In contrast compound 2 exhibited an increased rate of cell proliferation. The optical density of the control cell, is much higher that the plates for concentration 31.25 µg/ mL to 500 µg/ mL. However 2 cannot be discarded; this compound clearly shows that it possesses anti-hyperglycaemic properties and further studies are recommended.Item A comparative analysis of stability and structure-functional relationships of different xylanases(2013-07-30) Tabosa-Vaz, Sacha; Singh, Suren; Permaul, Kugen; Kumar, AjitA comparative thermostability analysis of different partially purified xylanases from Rhodothermus marinus, Bacillus halodurans, Thermomyces lanuginosus and Pulpzyme HC was studied using differential scanning fluorometry (DSF), fluorescence spectroscopy and circular dichroism (CD). The R. marinus xylanase was found to have an optimum temperature and pH of 90oC and 6 respectively while the B. halodurans xylanase was optimally active at 70oC and a broad range of alkaline pH of 8 - 10. The commercially available xylanase from T. lanuginosus showed optimal activity at 50oC and pH 7 while the Novozyme xylanase Pulpzyme HC showed optimal activity at 60oC and pH 7. Fluorescence spectroscopy monitored the microenvironment and fluorescence emission of Trp residues. In their native folded state, Trp are generally located in the core of the protein but during unfolding they become exposed. The fluorescence changes as the enzyme undergoes denaturation due to conformational changes and exposure of Trp residues. Differential scanning fluorometry (DSF) monitors thermal unfolding of proteins in the presence of a fluorescent dye such as Spyro Orange. A wide range of buffers were tested for their ability to increase the xylanase stability. T. lanuginosus had the greatest increase in melting temperature with 0.73M Bis Tris pH 6.5 and peaked highest at 78°C. The B. halodurans xylanase exhibited high pH stability (pH 4-10) and exhibited very little change in melting temperature, from 74°C-77°C over the twenty four different conditions. The R. marinus xylanase had no increase in melting temperature showing a maximum melting temperature of 90oC. Circular dichroism (CD) measures unequal absorption of right- and left-handed circularly polarized light by the molecule. The xylanase from R. marinus exhibited the lowest ΔG of 34.71kJ at 90°C as was expected. The B. halodurans xylanase showed a much higher ΔG of -52.71 at its optimum temperature of 70°C when compared with the xylanases from R. marinus and T. lanuginosus. When comparing the three xylanases activities at 70°C, it can be seen that the B. halodurans xylanase exhibited a lower relative activity then both R. marinus and T. lanuginosus xylanases. All three techniques offered different information on the structure and function relationship. Fluorescence spectroscopy, the change in conformation due to fluorescence emission as a result of increased temperature and salt concentrations. DSF, optimal conditions for increased stability and activity at higher temperatures and CD, conformational changes, the fraction of folded protein and change in Gibbs free energy over a range of temperature.