Aradi J, Zsindely A, Kiss A, Szabolcs M, Schablik M
Prep Biochem. 1982;12(2):137-51. doi: 10.1080/00327488208065558.
The purification of Neurospora crassa myo-inositol-1-phosphate synthase (EC 5.5.1.4) was studied by affinity chromatography using the substrate (glucose-6-phosphate), the inhibitor (pyrophosphate), the coenzyme (NAD+) and the coenzyme analogues (5'AMP and Cibacron Blue F3G-A) of the enzyme as adsorbents attached to agarose gel. Myo-inositol-1-phosphate synthase could be separated completely from the contaminating substance, glucose-6-phosphate dehydrogenase (EC 1.1.1.49), on Blue Sepharose CL-6B and on pyrophosphate-Sepharose. The purified enzyme had a specific activity of 16 400 U/mg. The sodium dodecyl sulfate/polyacrylamide gel electrophoresis of the 60 micrograms of this purified enzyme gave a homogenous band. The enzyme was found to be composed of four identical subunits having a molecular weight of 65 000.
利用酶的底物（6-磷酸葡萄糖）、抑制剂（焦磷酸）、辅酶（NAD⁺）以及辅酶类似物（5'-AMP和Cibacron Blue F3G-A）作为附着于琼脂糖凝胶的吸附剂，通过亲和色谱法研究了粗糙脉孢菌肌醇-1-磷酸合酶（EC 5.5.1.4）的纯化过程。肌醇-1-磷酸合酶能够在Blue Sepharose CL-6B和焦磷酸-琼脂糖上与污染物6-磷酸葡萄糖脱氢酶（EC 1.1.1.49）完全分离。纯化后的酶比活性为16400 U/mg。对60微克该纯化酶进行十二烷基硫酸钠/聚丙烯酰胺凝胶电泳得到一条均一的条带。发现该酶由四个分子量为65000的相同亚基组成。
