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Abstract This research dealt with optimization of the cyclosporine A (CyA) production by two methods; the first one was an optimization strategy based on statistical designs using Fusarium oxysporum in a submerged culture. A 2-level Plackett-Burman design was used to screen the most effective environmental and nutritional parameters on CyA production. Among the ten tested parameters, glycine concentration and incubation period were selected, owing to their significant positive effect on CyA production. A central composite rotatable design was adopted to acquire the best process parameters on both buffered & unbuffered conditions. Optimal combination of the major media constituents for CyA production was evaluated using SAS JMP 8 NULL program, resulting in a maximum CyA yield of 654.76 mg/L. The second one was the immobilization of Fusarium oxysporum spores through the use of Calcium alginate as an immobilization matrix. Different weights of spore inoculated beads (1, 2, 3, 4 g/50ml medium) and repeated utilization of the immobilized fungus were tested. Using of 3 g beads/flask resulted in maximum production of CyA (605.952 mg/L) in the first cycle. The immobilized spore beads were found to be unstable for the second cycle. . |