Purification and characterization of a phytase from Klebsiella terrigena.
A cytoplasmatic phytase was purified about 410-fold to apparent homogeneity with a recovery of 28%. The enzyme is induceable under carbon limitation in the presence of phytate. It behaves as a monomeric protein of a molecular mass of about 40 kDa. The phytase is rather specific for phytate and exhibits optimal conditions for phytate degradation at pH 5.0 and 58 degrees C. Kinetic parameters for the hydrolysis of Na phytate are KM 300 microM and kcat 180 s-1 at 35 degrees C and pH 5.0. Phytate is hydrolyzed in a stepwise manner; the penta- and tetrakisphosphate were identified as I(1,2,4,5,6)P5 and I(1,2,5,6)P4. Consequently, this enzyme is a 3-phytase (EC 3.1.3.8).
Greiner
R
R
Centre for Molecular Biology, Federal Research Centre for Nutrition, Karlsruhe, Federal Republic of Germany.
Haller
E
E
Konietzny
U
U
Jany
K D
KD
D010832:6-Phytase / Q000302:isolation & purification* / Q000378:metabolism; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002413:Cations, Divalent / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007709:Klebsiella / Q000201:enzymology*; D008970:Molecular Weight; D010833:Phytic Acid / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D001426:Bacterial Proteins; D002413:Cations, Divalent; D010833:Phytic Acid; D010832:6-Phytase