AFFINITY-CHROMATOGRAPHY PURIFICATION OF ALKALINE-PHOSPHATASE FROM CALF INTESTINE
AFFINITY-CHROMATOGRAPHY PURIFICATION OF ALKALINE-PHOSPHATASE FROM CALF INTESTINE
复制标题
DOI：
10.1042/bj1510291
复制
发表时间：
1975-01-01
影响因子：
4.1
通讯作者：
PIETTA, PG
中科院分区：
文献类型：
--
作者：
BRENNA, O;PERRELLA, M;PIETTA, PG
用琼脂糖亲和层析法从小牛肠粘膜中纯化出碱性磷酸酶粗品（EC 3.1.3.1）。制备了三种生物特异性吸附剂用于层析，并且用与来自4-（对氨基苯偶氮）苯胂酸的重氮盐偶联的酪胺基-琼脂糖衍生物获得了最佳结果。在不存在Pi的情况下，碱性磷酸酶是亲和柱保留的唯一酶。在pH10.0的磷酸缓冲液中，以5.5mM-对硝基苯磷酸为底物，酶的比活力为1200单位/mg蛋白。
A crude preparation of alkaline phosphatase (EC 3.1.3.1) from calf intestinal mucosa was purified by affinity chromatography on Sepharose-bound derivatives of arsanilic acid, which was found to be a competitive inhibitor of the enzyme. Three biospecific adsorbents were prepared for the chromatography, and the best results were obtained with a tyraminyl-Sepharose derivative coupled with the diazonium salt derived from 4-(p-aminophenylazo)phenylarsonic acid. Alkaline phosphatase was the only enzyme retained by the affinity column in the absence of Pi. The enzyme eluted by phosphate buffer had a specific activity of about 1200 units per mg of protein at pH 10.0, with 5.5mM-p-nitrophenyl phosphate as the substrate.