Purification and characterization of a phytase from Pseudomonas syringae MOK1

• Published in: Current microbiology Vol. 47; no. 4; pp. 290 - 294
• Main Authors: Cho, Jaie Soon, Lee, Chang Whan, Kang, Seung Ha, Lee, Jae Cheon, Bok, Jin Duck, Moon, Yang Soo, Lee, Hong Gu, Kim, Sung Chan, Choi, Yun Jaie
• Format: Journal Article
• Language: English
• Published: New York, NY Springer 01-10-2003; Springer Nature B.V
• Subjects: 6-Phytase - isolation & purification; 6-Phytase - metabolism; Animal Feed; Anion exchange; Bacteria; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Cadmium - metabolism; Chromatography, Ion Exchange - methods; Copper - metabolism; Dietary Fiber - metabolism; Edetic Acid - metabolism; Electrophoresis, Polyacrylamide Gel; Enzyme Inhibitors - analysis; Enzyme Stability; Enzymes; Fundamental and applied biological sciences. Psychology; Glycine max - metabolism; Hydrogen-Ion Concentration; Manganese - metabolism; Metabolism. Enzymes; Microbiology; Molecular Weight; Phosphates - metabolism; Phytic Acid - metabolism; Pseudomonas syringae; Pseudomonas syringae - enzymology; Sodium; Soybeans; Substrate Specificity; Temperature; Wheat bran; Pseudomonadales; Phosphates; Purification; Enzyme; 6-Phytase; Phosphoric monoester hydrolases; Esterases; Ion exchange chromatography; Characterization; Enzymatic activity; Pseudomonas syringae; Substrate specificity; Bacteria; Hydrolases; Pseudomonadaceae
• DOI: 10.1007/s00284-002-3966-4
• EISSN: 1432-0991
• ISSN: 0343-8651

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.