PURIFICATION AND CHARACTERIZATION OF TRANSGLUTAMINASE FROM WALLEYE POLLACK LIVER

Transglutaminase (TGase, EC 2.3.2.13) from walleye pollack Theragra Ch alcogramma liver was purified to electrophoretical homogeneity by Q-Se pharose and S-Sepharose chromatographies. The purified enzyme of 0.34 mg was obtained from 15 g of liver tissue and 591-fold purification wa s achieved from the liver extract. The molecular weight was estimated to be 77 kDa by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for monodansyl cadaverine incorporation to N,N'-dimet hylated casein were 9.0 and 50 degrees C, respectively. The purified e nzyme required Ca2+ above 3 mM for the maximum activity, and Sr2+ also fully activated the enzyme. The activity was inhibited by sulfhydryl reagent, suggesting this enzyme was a thiol enzyme, the same as mammal ian TGases. By this purified TGase, the gelation of myosin B solution was catalyzed, possibly through the polymerization of myosin heavy cha ins.