Purification and characterization of organic solvent and detergent stable protease isolated from marine Saccharopolyspora sp. A9: Application of protease for wound healing

Abstract

An extracellular organic solvent stable protease producing strain was isolated from west coast of India and identified as Saccharopolyspora sp. A9. The protease was purified from the culture supernatant to homogeneity using ammonium sulfate precipitation, DEAE Cellulose and Sephadex G-200 gel filtration with a 23.28-fold increase in specific activity having approximately 32kDa molecular weight. The enzyme stability was investigated over broad range of pH, temperature, salts, different organic solvents, commercial detergent and surfactants. In present study, carbopol 940 gel containing protease enzyme was developed for evaluation of its wound healing activity by excision wound model. The enzyme was stable in pH range of 8.0–12.0 with optimum of pH 10.0, whereas thermostable nature of enzyme was suggested by its stability over 70 and 80°C. In addition, enzyme showed excellent stability with a wide range of organic solvents having logP−2.0 and below 2.0. In excision wound model wounds were found to contract to 80.40% (p<0.001) in first nine days at higher rate with formulation containing 300U/g of protease enzyme. Therefore, topical protease formulation offers promise for its pharmaceutical use in wound healing and also proves its application in protein biosynthesis in non-aqueous media.