Characterization, gene cloning, and sequencing of a fungal phytase, PhyA, from Penicillium oxalicum PJ3.
A phytase from Penicillium oxalicum PJ3, PhyA, was purified near to homogeneity with 427-fold increase in specific phytase activity by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatographies. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis of the purified enzyme indicated an estimated molecular mass of 65 kD. The optimal pH and temperature of the purified enzyme were pH 4.5 and 55°C, respectively. The enzyme activity was strongly inhibited by Ca(2+), Cu(2+), Zn(2+), and phenylmethylsulfonyl fluoride (PMSF). The Km value for sodium phytate was 0.545 mM with a Vmax of 600 U/mg of protein. The phyA gene was cloned, and it contains an open reading frame of 1,383 with a single intron (118 bp), and encodes a protein of 461 amino acids.
a Department of Nano-Bioengineering , Incheon National University , Incheon , Korea.
Seung Ho
SH
Lee
Jaiesoon
J
Cho
Jinduck
J
Bok
Seungha
S
Kang
Yunjaie
Y
Choi
Peter C W
PC
Lee
D010832:6-Phytase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002118:Calcium / Q000494:pharmacology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003300:Copper / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D010833:Phytic Acid / Q000378:metabolism; D013696:Temperature; D015032:Zinc / Q000494:pharmacology
D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D010664:Phenylmethylsulfonyl Fluoride; D003300:Copper; D010833:Phytic Acid; D010832:6-Phytase; D015032:Zinc; D002118:Calcium
Penicillium oxalicum; phytase; purification; sodium phytate