Purification and characterization of a phytase from Pseudomonas syringae MOK1.
A phytase (EC 3.1.3.8) from Pseudomonas syringae MOK1 was purified to apparent homogeneity in two steps employing cation and an anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The optimal activity occurred at pH 5.5 and 40 degrees C. The Michaelis constant (Km) and maximum reaction rate (Vmax) for sodium phytate were 0.38 mM and 769 U/mg of protein, respectively. The enzyme was strongly inhibited by Cu2+, Cd2+, Mn2+, and ethylenediaminetetraacetic acid (EDTA). It showed a high substrate specificity for sodium phytate with little or no activity on other phosphate conjugates. The enzyme efficiently released orthophosphate from wheat bran and soybean meal.
Cho
Jaie Soon
JS
School of Agricultural Biotechnology, College of Agriculture and Life Science, Seoul National University, 441-744, Suweon, Korea.
Lee
Chang Whan
CW
Kang
Seung Ha
SH
Lee
Jae Cheon
JC
Bok
Jin Duck
JD
Moon
Yang Soo
YS
Lee
Hong Gu
HG
Kim
Sung Chan
SC
Choi
Yun Jaie
YJ
D010832:6-Phytase / Q000302:isolation & purification* / Q000378:metabolism*; D000821:Animal Feed; D002104:Cadmium / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000379:methods; D003300:Copper / Q000378:metabolism; D004043:Dietary Fiber / Q000378:metabolism; D004492:Edetic Acid / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000032:analysis; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000378:metabolism; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D010833:Phytic Acid / Q000378:metabolism; D044224:Pseudomonas syringae / Q000201:enzymology*; D013025:Glycine max / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D004043:Dietary Fiber; D004791:Enzyme Inhibitors; D010710:Phosphates; D002104:Cadmium; D008345:Manganese; D003300:Copper; D010833:Phytic Acid; D004492:Edetic Acid; D010832:6-Phytase