Statistical media optimization for enhanced production of fibrinolytic enzyme from newly isolated Proteus penneri SP-20

Thrombolytic enzymes are invaluable for combating cardiovascular diseases. In this study a thorough isolation resulted, a potent fibrinolytic enzyme producer which was identified as Proteus penneri SP-20 using 16S rRNA sequencing. It secretes extracellular protease possesing fibrinolytic activity which was confirmed through the analysis of fibrin degradation by SDS-PAGE, phase contrast microscopy and fibrin zymography. Response surface methodology is used for media optimization using newly isolated P. penneri SP-20. Plackett Burman (PB) design was employed for selection of critical media constituents while; further optimization was carried out using Central composite rotatable design (CCRD). Casein, yeast extract, maltose and MnCl2·4H2O were critical medium components selected on the basis of PB design. Optimized media components obtained by CCRD were casein; 12.58 g l−1, yeast extract; 17.27 g l−1, maltose; 1.65 g l−1, MnCl2·4H2O; 0.564 g l−1. As compared with initial enzyme activity of 38.19 Units CCRD yielded 796.82 Unit activity, increasing the overall production of fibrinolytic protease by 20.86 fold. Hence the model is reliable for process optimization.