Isolation and characterization of catalase from Penicillium chrysogenum.
Isolation and characterization of catalase from Penicillium chrysogenum.
复制标题
产黄青霉过氧化氢酶的分离和表征。
DOI：
10.1016/0021-9673(92)85542-2
复制
发表时间：
1992
影响因子：
4.1
通讯作者：
J. Porath
中科院分区：
文献类型：
--
作者：
G. Chaga;A. Medin;S. G. Chaga;J. Porath
用Cu（Ⅱ）-螯合Sepharose Fast Flow固定化金属离子亲和层析（IMAC），从产黄青霉（Penicillium chrysogenum）粗品中分离出过氧化氢酶。以89%的产率获得了色谱和电泳均相的酶。IMAC被认为是上级优于离子交换，疏水相互作用，尺寸排阻和刀豆球蛋白A亲和层析。分析色谱法和制备色谱法的色谱图基本相同。测定了等电点、分子量（超离心）、氨基酸组成、碳水化合物含量和亚基组织。计算表观米氏常数KM和叠氮化物竞争常数Ki，结果分别为59 μ M和6.1 μM。
Catalase from a crude preparation ofPenicillium chrysogenumwas isolated in a single chromatographic step by immobilized metal ion affinity chromatography (IMAC) on Cu(II)-Chelating Sepharose Fast Flow. A chromatographically and electrophoretically homogeneous enzyme was obtained in 89% yield. IMAC was found to be superior to ion-exchange, hydrophobic interaction, size-exclusion and concanavalin A affinity chromatography. Analytical and preparative chromatography essentially the same chromatograms. Isoelectric point, molecular weight (by ultracentrifugation), amino acid composition, carbohydrate content and subunit organization were determined. The apparent Michaelis-Menten constant,KM, and the azide competitor constant,Ki, were calculated and found to be 59 μMand 6.1 μM, respectively.