AFFINITY THERMOPRECIPITATION OF LACTATE-DEHYDROGENASE AND PYRUVATE-KINASE FROM PORCINE MUSCLE USING EUDRAGIT BOUND CIBACRON BLUE

Cibacron blue coupled to Eudragit S 100 was used for isolation of lactate dehydrogenase and pyruvate kinase from crude porcine muscle extract by the technique of affinity thermoprecipitation. The polymer preparation with an immobilized dye content of about 195 mg g(-1) dry weight was precipitated at 40 degrees C in the presence of 50 mM Ca2+ ions, and was made soluble at room temperature in the absence of the calcium ions. Isolation of the enzymes from the crude extract by co-precipitation with Eudragit-Cibacron blue was studied at two loadings of the extract. At a low load, both the enzymes were precipitated and were desorbed sequentially by treatment with increasing concentrations of salt. Of the two enzymes, mainly lactate dehydrogenase was precipitated from the high load of the crude extract and was recovered with a higher purity.