PURIFICATION AND PARTIAL CHARACTERIZATION OF ALKALINE-PHOSPHATASE FROM CALF INTESTINE MUCOSE

The present study describes a simple method for obtaining relative hig hly purified enzyme alkaline phosphatase (AP) from fresh mucose of cal f small intestine. The purification procedure involved homogenization in 0.015 mol.l-1 Tris-HCl buffer pH 7.9, followed ammonium sulphate pr ecipitation, precipitation with acetone, solubilisation thus obtained material with Triton X-100, gel filtration on Sephadex G-100 column an d anion-exchange chromatography and DEAE cellulose column. Native PAGE electrophoresis as wel as FPLC system revealed considerable purity of thus obtained AP. The purified AP exhibited an apparent molecular wei ght of 105,000 as judged by SDS-PAGE electrophoresis. The yield of the method described here was about 23 %. The AP obtained by us is sensit ive to exposing temperature aver 37-degrees-C. Our results confirmed c onsiderable stimulation of AP activity by Mg2+ and Mn2+ ions.