Extraction, Purification and Characterization of Transglutaminase From Some Plants

Abstract

The present study aimed to Isolate trans-glutaminase EC:2.3.2.13 from some plants sources and Purified it and Studied it’s Charctarestics as well as it’s practical applications in the production of sausage.The enzyme was extracted from four types of plants (rosemary, chard, radish, arugula) using nine extraction solutions that included distilled water,Sodium chloride 3% solution, sodium chloride 5% solution, sodium phosphate solution 0.1 M and an pH 6.5, sodium phosphate 0.1 M and an pH of 7.5, Tris - Hcl solution 0.2 M and pH 7, Tris - Hcl solution 0.2 M and an pH 8, Tris - Hcl solution 0.1 M and an pH 7 and Tris - Hcl solution 0.1 M and an pH 8 in order to find out the best source of enzyme and the best extraction solution. chard was the best source of enzyme compared with other sources, Tirs -HCl 0.1M, pH 8 solution was the best extraction solution which gave the highest specific activity 8.104 unit/mg.Protein content for the crude enzyme extracts were concentrated using saturated ammonium sulfate in arrange 20-60%, Dialysis was done using distilled water. Then, the purification steps of the enzyme were completed using the gel filtration in the Sephadex G-100 Purification Folds 13.91 time and the yield was 20.04% %. Electrophoresise process using poly acryl amid gel in the absence of SDS observe the presence of one protein band which indicates the complete purification of transglutaminase. transglutaminase molecular weight was 42,660 Dalton when it was evaluated using poly acryl amide electrophoresis in the presence of SDS. the optimum pH for enzyme activity and enzyme stability was 7, while the optimum temperature for enzyme activity was 55°C, and the optimum temperature for enzyme stability was between 25-45°C.