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Membrane bioreactor production of lignin and manganese peroxidase
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The white-rot fungus (WRF), Phanerochaete chrysosporium, is a well known microorganism which produces ligninolytic enzymes. These enzymes can play a major role in the bioremediation of a diverse range of environmental aromatic pollutants present in industrial effluents. Bioremediation of aromatic pollutants using ligninolytic enzymes has been extensively researched by academic, industrial and government institutions, and has been shown to have considerable potential for industrial applications. Previously the production of these enzymes was done using batch cultures. However, this resulted in low yields of enzyme production and therefore an alternative method had to be developed. Little success on scale-up and industrialisation of conventional bioreactor systems has been attained due to problems associated with the continuous production of the pollutant degrading enzymes. It was proposed to construct an effective capillary membrane bioreactor, which would provide an ideal growing environment to continuously culture an immobilised biofilm of P; chrysosporium (Strain BKMF-1767) for the continuous production of the ligninolytic enzymes, Lignin(LiP) and Manganese(MnP) Peroridase. A novel membrane gradostat reactor (MGR) was shown to be superior to more conventional systems of laboratory scale enzyme production (Leukes et.al., 1996 and Leukes, 1999). This concept was based on simulating the native state ofthe WRF, which has evolved on a wood-air interface and involved irnmobilisng the fungus onto an externally skinless ultrafiltration membrane. The MGR however, was not subjected to optimisation on a laboratory scale. The gradostat reactor and concept was used in this work and was operated in the deadend filtration mode. The viability of the polysulphone membrane for cultivation of the fungus was investigated. The suitability of the membrane bioreactor for enzyme production was evaluated. The effect of microbial growth on membrane pressure and permeability was monitored. A possible procedure for scaling up from a single fibre membrane bioreactor to a multi-capillary system was evaluated. Results indicated that the polysulphone membrane was ideal for the cultivation of P chrysosporium, as the micro-organism was successfully immobi1ised in the macrovoids of the membrane resulting in uniform biofilm growth along the outside of the membrane. The production of Lignin and Manganese Peroxidase was demonstrated. The enzyme was secreted and then transported into the permeate without a rapid decline in activity. Growth within the relatively confined macrovoids of the membrane contributed to the loss of membrane permeability. A modified Bruining Model was successfully applied in the prediction of pressure and permeability along the membrane The study also evaluated the effect of potential1y important parameters on the production of the enzymes within the membrane bioreactor. These parameters include air flow (Ch concentration), temperature, nutrient flow, relative redox potential and nutrient concentrations A sensitivity analyses was performed on temperature and Ch concentration. The bioreactor was exposed to normal room temperature and a controlled temperature at 37°C. The reactors were then exposed to different O2 concentration between 21% and 99"10. It was found that the optimum temperature fur enzymes production is 3TJC. When oxygen was used instead of air, there was an increase in enzyme activity. From the results obtained, it was clear that unique culture conditions are required for the production of LiP and MnP from Phanerochaete chrysosporium. These culture conditions are essential fur the optimisation and stability of the bioreactor.