Statistical medium optimization for the production of collagenolytic protease by Pseudomonas sp SUK using response surface methodology

Pseudomonas sp. SUK producing an extracellular collagenolytic protease was isolated from soil samples from meat and poultry industrial area based in Kolhapur, India. Response surface methodology was employed for the optimization of different nutritional parameters influencing production of collagenolytic protease by newly isolated Pseudomonas sp. SUK in submerged fermentation. Initial screening of production parameters was performed using Plackett-Burman design and the variables with statistically significant effects on collagenolytic protease production were identified as gelatin, peptone, and K2HPO4. Further, optimization by response surface methodology (RSM) using Central Composite Design showed optimum production of collagenolytic protease with 12.05 g L-1 of gelatin, 12.26 g L-1 of peptone and 1.29 g L-1 of K2HPO4. Collagenolytic protease production obtained experimentally has very close agreement with the model prediction value and the model was proven to be adequate. The statistical optimization by response surface methodology upsurges collagenolytic protease yield by 2.9 fold, hence the experimental design is effective towards process optimization. Moreover, ammonium sulphate precipitated, partially purified enzyme has shown to cleave collagen from bovine achilles tendon, which was observed by phase contrast microscopy, and SDS-PAGE. Hence, extracellular collagenolytic protease of Pseudomonas sp. SUK could have considerable potential for industrial as well as medical applications.