Purification of galactose oxidase from Dactylium dendroides by affinity chromatography on melibiose-polyacrylamide.
Purification of galactose oxidase from Dactylium dendroides by affinity chromatography on melibiose-polyacrylamide.
复制标题
通过蜜二糖-聚丙烯酰胺亲和层析纯化来自 Dactylium dendroides 的半乳糖氧化酶。
DOI：
10.1016/0003-9861(88)90645-5
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发表时间：
1988
影响因子：
3.9
通讯作者：
Bhavanandan,VP
中科院分区：
文献类型：
--
作者：
Kelleher,FM;Dubbs,SB;Bhavanandan,VP
半乳糖氧化酶是已知将半乳糖和半乳糖胺的C-6羟甲基氧化成醛基的真菌酶。它已被广泛用于糖缀合物的研究，例如在去唾液酸糖蛋白的标记。我们已经开发了一种简单的亲和纯化半乳糖氧化酶使用蜜二糖-聚丙烯酰胺。这种亲和过程被用来纯化酶的硫酸铵沉淀的培养物dactylandendroides。含有蛋白酶和其他污染物的材料在缓冲液洗涤中洗脱。然后用含有0.1m d-岩藻糖或半乳糖的缓冲液从柱中特异性洗脱半乳糖氧化酶。使用这种方法，酶也被纯化，从商业样品的半乳糖氧化酶含有高蛋白水解活性。
Galactose oxidase is a fungal enzyme which is known to oxidize the C-6 hydroxymethyl of galactose and galactosamine to an aldehyde group. It has been widely used in glycoconjugate research, for example in the labeling of asialoglycoproteins. We have developed a simple affinity purification for galactose oxidase using melibiose-poly-acrylamide. This affinity procedure was used to purify the enzyme from ammonium sulfate precipitates of culture filtrates ofDactylium dendroides. The material containing proteases and other contaminants is eluted in the buffer wash. The galactose oxidase is then specifically eluted from the column with buffer containing 0.1m d-fucose ord-galactose. Using this procedure, the enzyme was also purified from commercial samples of galactose oxidase which contain high proteolytic activity.