Purification and properties of endopolygalacturonase i from stereum purpureum, a factor inducing silver-leaf symptoms on apple trees
Purification and properties of endopolygalacturonase i from stereum purpureum, a factor inducing silver-leaf symptoms on apple trees
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苹果树银叶症状诱导因子紫紫内切多半乳糖醛酸酶 i 的纯化及其特性
DOI：
10.1271/bbb1961.49.1111
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
K. Sawai
中科院分区：
文献类型：
--
作者：
K. Miyairi;T. Okuno;K. Sawai
采用CM-52和Sephadex G-100的柱层析技术，从紫体菌培养液中纯化出引起银叶症状的内聚半乳糖醛酸酶I [EC 3.2.1.15]。纯化后的酶经聚丙烯酰胺凝胶电泳和超离心均相。沉淀系数（S20, w）确定为3.21 S，凝胶过滤估计分子量为40000，sds -聚丙烯酰胺凝胶电泳估计分子量为41000，沉淀平衡估计分子量为44000。酶的等电点pH为8.5。酶的最佳pH为：三半乳糖醛酸3.5，四半乳糖醛酸4.0，果胶酸4.5。该酶在pH 4.0 ~ 9.0范围内稳定，温度高达70°C 30min。诱导苹果树银叶症状所需酶的用量为20μg/棵。
Endopolygalacturonase I [EC 3.2.1.15], the major component of endopolygalacturonases causing silver-leaf symptoms, was purified from culture liquids of Stereum purpureum by column chromatographies on CM-52 and Sephadex G-100. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis and ultracentrifugation. The sedimentation coefficient (S20, w) was determined to be 3.21 S, and the molecular weight was estimated to be 40, 000 by gel filtration, 41, 000 by SDS-polyacrylamide gel electrophoresis and 44, 000 by sedimentation equilibrium. The enzyme had an isoelectric point of pH 8.5. The optimal pH of the enzyme was 3.5 for trigalacturonic acid, 4.0 for tetragalacturonic acid, and 4.5 for pectic acid. The enzyme was stable in the range of pH 4.0 to 9.0 and up to 70°C for 30min. The amount of the enzyme which was required to induce silver-leaf symptoms on apple trees was 20μg/tree.