Response surface methodology for optimization of medium components in submerged culture of Aspergillus flavus for enhanced heparinase production

Aims: Aim of the study was to develop a medium for optimal heparinase production with a strain of Aspergillus flavus (MTCC-8654) by using a multidimensional statistical approach. Methods and Results: Statistical optimization of intracellular heparinase production by A. flavus, a new isolate, was investigated. Plackett-Burman design was used to evaluate the affect of medium constituents on heparinase yield. The experimental results showed that the production of heparinase was dependent upon heparin, the inducer; chitin, structurally similar to heparin and NH4NO3, the nitrogen source. A central composite design was applied to derive a statistical model for optimizing the composition of the fermentation medium for the production of heparinase enzyme. The optimum fermentation medium consisted of (g l(-1)) Mannitol, 8 center dot 0; NH4NO3, 2 center dot 5; K2HPO4, 2 center dot 5; Na2HPO4, 2 center dot 5; MgSO4.7H(2)O, 0 center dot 5; Chitin, 17 center dot 1; Heparin, 0 center dot 6; trace salt solution (NaMoO4.2H(2)O, CoCl2.6H(2)O, CuSO4.5H(2)O, FeSO4.7H(2)O, CaCl2), 10(-4) mol l(-1). Conclusions: A 2 center dot 37-fold increase in heparinase production was achieved in economic and effective manner by the application of statistical designs in medium optimization. Significance and Impact of the Study: Heparinase production was doubled by statistical optimization in a cost-effective manner. This heparinase can find application in pharmaceutical industry and for the generation of low-molecular-weight heparins, active as antithrombotic and antitumour agents.