Browsing by Author "Salawudeen, Morufat Tolani"

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    Production and purification of xylanase from a fungus isolated from a paper industry solid waste dumpsite in Osogbo,Osun State, Nigeria
    (Department of Microbiology, Faculty of Science, Obafemi Awolowo Universty, 2023) Salawudeen, Morufat Tolani
    This study isolated and screened for xylanolytic fungi from a paper industry solid waste dumpsite. It also identified the selected fungus and then optimized the process parameters for xylanase production from selected fungus and to produced, purified as well as characterized xylanase produced from fungi isolated from a paper industry solid waste dumpsite. These were with a view to providing useful information on the potential utilization of agricultural waste as a substrate for the production of xylanase to fulfill various industrial needs. Soil samples were collected aseptically from four different location at a paper industry waste dumpsite in Osogbo, Osun State. The samples collected was processed for isolation using spread plate method and standard methods were used for characterizing and identifying the fungal isolates, which were subsequently subjected to screening using both rapid plate assay method and submerged fermentation for the process of producing xylanase. The fungus with the best xylanase activity was selected for enzyme production, purification and characterization. The best optimal conditions for xylanase synthesis by the best producing fungus were determined by varying the nitrogen sources, pH, carbon sources, temperature, inoculum volumes and different concentrations of the carbon and nitrogen sources for media composition. The best optimal condition were used for bulk synthesis of xylanase to produce the crude enzyme. The crude enzyme was partially purified using a one step purifications method of ion exchange chromatography using CM-Sephadex C-50. The effect of pH, temperature, metal ions, heat stability were studied and the kinetic parameters (Km and Vmax) of the partially purify xylanase were determined. The selected fungus, SD5, has a zones of hydrolysis and highest enzyme activity of about 12 mm and 212.30 Units/ml/min respectively from the 32 isolates obtained from the samples collected from the paper waste dumpsites after been screened. The fungus SD5 was molecularly characterized and identified as Aspergillus oryzae strain KhA0707 with 98% identity. With an inoculum amount of 1.0 ml and 4 days of fungal growth, the enzyme production reached its peak. The best optimal temperature and pH of production was 30°C and pH 6.0 respectively, while the best nitrogen and carbon sources are peptone and xylan respectively. The partially purified enzyme showed that specific activity was 5437.35 units/mg with purification fold of 12.33. Characterization of the partially purified xylanase showed that the optimum activity occurred at temperature and pH of 55 °C and 6.0 respectively. The enzyme kinetic Vmax value is 161.290 units/ml/min and Km value of 0.6129 mg/ml. EDTA totally inhibited the activity of the partially purified xylanase. The enzyme activity was slightly inhibited by Ca2+, Na+, even at 1 mM while K+, Cu2+, Al3+, urea and Mecarptopyruvate promote the xylanase activity. The findings of this research showed that a paper waste dumpsite is also a potential source for xylanolytic fungus. The isolated Aspergillus oryzae employed for xylanase production in this study was able to generate a significant quantity of xylanase, which will be useful in protein hydrolysis and industries.