酵母乌头酸酶。II. 酵母乌头酸酶的结晶及一般性质

The aconitase of yeast. II. Crystallization and general properties of yeast aconitase.

作者信息

Suzuki T, Yamazaki O, Nara K, Akiyama S, Nakao Y

出版信息

J Biochem. 1975 Feb;77(2):367-72. doi: 10.1093/oxfordjournals.jbchem.a130734.

DOI:10.1093/oxfordjournals.jbchem.a130734

Abstract

Yeast aconitase [citrate (isocitrate) hydro-lyase, ED 4.2.1.3], inductively formed by Candida iipolytica in the presence of fluoroacetate, was purified approximately 100-fold by Sephadex G-100 gel filtration and DEAE-Sephadex column chromatography, yielding dark-brown needle crystals. The crystalline aconitase was homogenious as judged by polyacrylamide gel electrophoresis and sedimentation by ultracentrifugation. The enzyme showed maximal activity at pH 8.0 and at 55 degrees. It has an S20, W of 5.03 S, a molecular weight of 68,500 and an isolectric point of pH 4.2. The presence of 2.10 moles of iron per mole of the enzyme was demonstrated by atomic absorption spectroscopy.

摘要

酵母乌头酸酶[柠檬酸（异柠檬酸）水解酶，EC 4.2.1.3]，由解脂假丝酵母在氟乙酸存在下诱导形成，通过Sephadex G - 100凝胶过滤和DEAE - Sephadex柱色谱法纯化约100倍，得到深棕色针状晶体。通过聚丙烯酰胺凝胶电泳和超速离心沉降判断，结晶乌头酸酶是均一的。该酶在pH 8.0和55℃时表现出最大活性。它的沉降系数S20,W为5.03 S，分子量为68,500，等电点为pH 4.2。通过原子吸收光谱法证明每摩尔酶中存在2.10摩尔铁。