Lee Seung Ho, Cho Jaiesoon, Bok Jinduck, Kang Seungha, Choi Yunjaie, Lee Peter C W
a Department of Nano-Bioengineering , Incheon National University , Incheon , Korea.
Prep Biochem Biotechnol. 2015;45(4):336-47. doi: 10.1080/10826068.2014.923446.
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.
从草酸青霉PJ3中纯化得到一种植酸酶PhyA，通过硫酸铵沉淀、凝胶过滤和离子交换色谱法，其比植酸酶活性提高了427倍，纯度接近均一。十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和纯化酶的酶谱分析表明，其估计分子量为65 kD。纯化酶的最适pH和温度分别为pH 4.5和55°C。该酶活性受到Ca(2+)、Cu(2+)、Zn(2+)和苯甲基磺酰氟（PMSF）的强烈抑制。植酸钠的Km值为0.545 mM，Vmax为600 U/mg蛋白质。克隆了phyA基因，它包含一个1383个碱基的开放阅读框，带有一个单一内含子（118 bp），编码一个461个氨基酸的蛋白质。
