PURIFICATION AND CHARACTERIZATION OF PHYTASE INDUCED IN TOMATO ROOTS UNDER PHOSPHORUS-DEFICIENT CONDITIONS

Phytase (myo-inositol hexakisphosphate phosphohydrolase; EC 3.1.3.8) w as purified from roots of tomato plants grown under phosphorus-deficie nt conditions using five purification schemes, The phytase was success fully separated from the major acid phosphatase to an electrophoretic homogeneity, The native molecular weight of this enzyme was estimated to be about 164 kD by Bio-Gel P-200 gel filtration, The molecular weig ht of the subunit on SDS-PAGE was approximately 82 kD, indicating that the native form of the enzyme was a homodimer, The isoelectric point of tomato phytase was about 5.5, The enzyme exhibited a high affinity for phytic acid (K-m=38 mu M), and was strongly inhibited by phosphate , molybdate and fluoride, Among other characteristics of tomato phytas e, the pH and temperature optima were 4.3 and 45 degrees C, respective ly, Tomato phytase contained a fairly high concentration of aspartic, glutamic acid and glycine residues.