[巨大内孢霉中多种形式的多聚磷酸磷酸水解酶的检测]
[Detection of multiple forms of polyphosphate phosphohydrolase from Endomyces magnusii].
作者信息
Afanas'eva T P, Uryson S O, Kulaev I S
出版信息
Biokhimiia. 1976 Jul;41(6):1078-86.
Labile polyphosphate phosphohydrolase from Endomyces magnusii is 27-fold purified by means of fractionation with ammonium sulphate, gel filtration on Sephadex G-75 and Biogel P-60 and chromatography on DEAE cellulose. Chromatography on DEAE Sephadex A-50, isoelelctric focusing and polyacrylamide gel electrophoresis of the enzyme preparation revealed 3 different fractions with polyphosphate phosphohydrolase activity (PPPH1, PPPH2 and PPPH3). Relative content of these fractions in E. magnusii cells is 30%, 55% and 15% respectively. Isoelectric points are: PPPH1--pH 5.1--5.2; PPPH2--pH 6.0--6.1; PPPH3--pH 6.3--6.4. PPPH1 and PPPH2 are found to be the most labile. PPPH3 is more stable under isolation procedure and storage. The fractions have similar molecular weight (48 000 +/- 3000).
来自大孢内孢霉的不稳定多聚磷酸磷酸水解酶通过硫酸铵分级分离、在葡聚糖凝胶G - 75和Biogel P - 60上的凝胶过滤以及在DEAE纤维素上的色谱法进行了27倍的纯化。对酶制剂进行DEAE - 葡聚糖A - 50色谱、等电聚焦和聚丙烯酰胺凝胶电泳，结果显示有3种具有多聚磷酸磷酸水解酶活性的不同组分（PPPH1、PPPH2和PPPH3）。这些组分在大孢内孢霉细胞中的相对含量分别为30%、55%和15%。等电点分别为：PPPH1——pH 5.1 - 5.2；PPPH2——pH 6.0 - 6.1；PPPH3——pH 6.3 - 6.4。发现PPPH1和PPPH2最不稳定。PPPH3在分离过程和储存中更稳定。这些组分具有相似的分子量（48000 ± 3000）。
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