title: Substrate specificity and pH dependence of homogeneous wheat germ acid phosphatase.
abstract: The broad substrate specificity of a homogeneous isoenzyme of wheat germ acid phosphatase (WGAP) was extensively investigated by chromatographic, electrophoretic, NMR, and kinetic procedures. WGAP exhibited no divalent metal ion requirement and was unaffected upon incubation with EDTA or o-phenanthroline. A comparison of two catalytically homogeneous isoenzymes revealed little difference in substrate specificity. The specificity of WGAP was established by determining the Michaelis constants for a wide variety of substrates. p-Nitrophenyl phosphate, pyrophosphate, tripolyphosphate, and ATP were preferred substrates while lesser activities were seen toward sugar phosphates, trimetaphosphate, phosphoproteins, and (much less) phosphodiesters. An extensive table of Km and Vmax values is given. The pathway for the hydrolysis of trimetaphosphate was examined by colorimetric and 31P NMR methods and it was found that linear tripolyphosphate is not a free intermediate in the enzymatic reaction. In contrast to literature reports, homogeneous wheat germ acid phosphatase exhibits no measurable carboxylesterase activity, nor does it hydrolyze phenyl phosphonothioate esters or phytic acid at significant rates.
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, Indiana 47907-1393.
authors.forename: R L
authors.initials: RL
authors.lastname: Van Etten
authors.forename: P P
authors.initials: PP
authors.lastname: Waymack
mesh_terms: D010832:6-Phytase / Q000378:metabolism; D000135:Acid Phosphatase / Q000378:metabolism*; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D013379:Substrate Specificity; D013816:Thermodynamics; D014908:Triticum / Q000201:enzymology*
chemical_list: D007527:Isoenzymes; D002265:Carboxylic Ester Hydrolases; D000135:Acid Phosphatase; D010832:6-Phytase