ISOLATION AND PURIFICATION OF TRANSGLUTAMINASE 1 USING BIOCHEMICAL TECHNIQUES.

Transglutaminase 1 catalyzes the creation of covalent bonds between proteins, play an essential role in various biological processes and industrial applications. The study aims to isolate and purify transglutaminase 1 from the blood serum of healthy individuals using numerous biochemical techniques. TGMs 1 are isolated and purified from the blood serum of healthy volunteers samples who were not smokers and had not taken any medications at the time of the sample collection. The results show that these techniques included precipitation with 65% ammonium sulfate, dialysis, and negative ion exchange chromatography, successfully separating a single prominent band with high activity using DEAE-cellulose. The enzyme activity recovery was estimated at approximately 33.01%. Subsequently, gel filtration using Sephadex G-100 revealed a single fraction with high TGM 1 activity. This fraction exhibited a purification factor of 9.09, with an estimated recovery of enzyme activity of around 29.6%. The isolated and purified TGM 1's approximate molecular weight was around 73,115 Daltons, as assessed through gel filtration chromatography with Sephadex G-100. The study indicated that the optimal conditions for the isolated and partially purified TGM 1 enzyme were a pH of 6.4 and a temperature of 37°C, using a concentration of 0.5 mmol/L of the substrate tetramethylbenzidine. The results indicated that purified TGM1 may be an alternative to other sources.