PRODUCTION AND CHARACT RIZATION OF CRUD INTRAC LLUL R PHYTAS FROM R COMBINANT BACT RIA pEAS1AMP

Abstract

This research was aimed a producing a crude intracellular phy ase charac erized from recombinan bac eria.The recombinan bac eria (pEAS1AMP )was produced by way of ransforming pET-22b(+) +pEAS1 in o compe ent E.coli BL21 and E.coli BL21(DE3)cells.Crude in racellular phy ase produc ion was induced using 1,5 mM Isopropyl-β-D-thiogalactopyranosid (IPTG).Recombinant bac eria produc and enzyme ac ivi y es followed he Sajidan me hod.E.coli BL21(+)pEAS1 and E. coli BL21 (DE3)(+)pEAS1 recombinan bac eria showed grow h af er 20 hours and 10 hours of ransforma ion.Phy ase ac ivi y of E.coli BL21 (DE3)(+)+pEAS1 showed higher han hose of E.coli BL21(+)+pEAS1 .Crude intracellular phy ase of pEAS1AMP recombinan bac eria has an op imum ac ivi y a pH 5,40 o C,incuba ion period of 60 minutes,subs ra e concen ra ion of 2%,molecular weigh (MW)of 47.3 kDa,Km =15.91 u and Vm =2.41 u second.Mg 2+ ac s as a cofac or bu Fe 3+ (10 -4 M)ac s as an inhibi or. K ywords:bact ria r combinant pEAS1AMP,compet nt c lls,crud intrac llular phytase