Extraction and Purification of Ficin from Leaves of Ficus and Study it's Proteolytic Efficiency

Abstract

The present study aimed to extraction and purification ficin enzymewhich extracted from leaves of Ficus and study it's proteolytic efficiency. Manysolutions were used for extraction such as distilled water, sodium chloride (2 and 7%)solution ,Sodium Phosphate with citrate solution 0.1M and pH 7, Ethanol, coldacetone with citrate and phosphate solution and Tris-Hcl 0.1 M, pH 9 in order to findout the best extraction solution. Tris-HCl 0.1 M pH 9 solution was the best extractionsolution which gave the highest specific activity being 93.48unit / mg Protein . Proteincontent for the crude enzyme extracts was precipitated by gradual addition of solidammonium sulphate to final saturation of 20 -90% the activity being 688.95 unit /mg Purification Folds 6 time and 26.12% as a yield . Gel filtration followed bySephadex G-100 and ion exchange chromatography through DEAE Sephadex A-50columin were used to complete purification of ficin and tested proteolytic activity ineach step of purification which exhibit 2733.36, Purification Folds 23.90 time and15.43% as a yield. The purity of ficin was examined by poly acryl amide gelelectrophoresis.