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 NH(4)NO(3,) 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, 8.0; NH(4)NO(3), 2.5; K(2)HPO(4), 2.5; Na(2)HPO(4), 2.5; MgSO(4).7H(2)O, 0.5; Chitin, 17.1; Heparin, 0.6; trace salt solution (NaMoO(4).2H(2)O, CoCl(2).6H(2)O, CuSO(4).5H(2)O, FeSO(4).7H(2)O, CaCl(2)), 10(-4) mol l(-1).

CONCLUSIONS

A 2.37-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.