谷氨酸脱羧酶亲和层析介质制备

将 Sepharose 4B用环氧氯丙烷活化 ,接丁二胺臂 ,再用环氧氯丙烷活化 ,与谷氨酸的 N- 2氨基偶联。在 N ,N-二环己碳化二亚胺催化下 ,以接臂的胶与谷氨酸的 C- 5和 C- 1羧基基团偶联。结果表明 ,谷氨酸 N- 2氨基偶联和 C- 1羧基偶联的亲和介质对小麦谷氨酸脱羧酶有部分亲和作用 ,洗脱下的酶活低 ,而谷氨酸 C- 5羧基基团偶联的层析介质对小麦谷氨酸脱羧酶有完全亲和作用 ,洗脱的酶活高。谷氨酸能从亲和材料上洗脱掉脱羧酶 ,但它抑制酶活 ,而同等浓度的 4-氨基丁酸和谷氨酰胺梯度洗脱没有作用

Affinity chromatography materials for glutamate decarboxylase

Sepharose gels were activated with epichlorohydrin,spaced with putrescine and reactivated with epichlorohydrin.They were coupled with α-amino group of L-glutamate.Spaced gels were coupled with α-carboxylic and δ- carboxylic group of L-glutamate by the aid of N,N-dicyclohexlcarbodiimide respectively.The glutamate decarboxylase from wheat was completely absorbed by Glu Sepha rose with coupled δ-carboxylic group,partly absorbed by other affinity gels.Gl utamate could elute the enzyme but inhibit the enzyme activity.Glutamine and 4- aminobytrate could not elute the enzyme at the same concentration.