This research was aimed at producing a crude intracellular phytase characterized from recombinant
bacteria. The recombinant bacteria (pEAS1AMP) was produced by way of transforming pET-22b(+)
+pEAS1 into competent E. coli BL21 and E. coli BL21(DE3) cells. Crude intracellular phytase
production was induced using 1,5 mM Isopropyl-β-D-thiogalactopyranosid (IPTG). Recombinant
bacteria product and enzyme activity test followed the Sajidan method. E. coli BL21(+)pEAS1 and E.
coli BL21 (DE3)(+)pEAS1 recombinant bacteria showed growth after 20 hours and 10 hours of
transformation. Phytase activity of E. coli BL21 (DE3)(+)+pEAS1 showed higher than those of E. coli
BL21(+)+pEAS1. Crude intracellular phytase of pEAS1AMP recombinant bacteria has an optimum
activity at pH 5, 40o
C, incubation period of 60 minutes, substrate concentration of 2%, molecular weight
(MW) of 47.3 kDa, Km = 15.91 υM and Vm = 2.41 υM/second. Mg2+
acts as a cofactor but Fe3+
(10-4
M) acts as an inhibitor.
Keywords: bacteria recombinant pEAS1AMP, competent cells, crude intracellular phytas
