Optimization of microbial collagenolytic enzyme production by Bacillus subtilis subsp. Subtilis S13 using Plackett-Burman and response surface methodology

Bacillus subtilis subsp. subtilis S13, a new isolate from soil is found to be the collagenase producing bacteria. The results showed that gelatin concentration, initial pH, and incubation time were accounted for significant factors from PB design and then applied for a central composite design (CCD) under response surface methodology (RSM) for optimization of significant factors were performed. The optimum parameters for the enhancing gelatinase production through CCD and response surface methodology were 19 g/l of pork gelatin, initial pH 6.16 for culture medium, and 59 h of incubation time, which provided the predicted maximum collatinolytic activity of 65.36 U/ml. This condition allowed approximately increasing 4-folds as compared to un-optimized condition (15 U/ml). The obtained optimal activity of this enzyme might be expressed the potential collagenase for several applications including meat tenderizing enzyme.