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谷氨酸脱羧酶亲和层析介质制备
范军1, 李培金1, 李 纯1
安徽农业大学 生物技术研究中心
摘要:
将Sepharose’r4B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。
关键词:  谷氨酸  谷氨酸脱羧酶  亲和层析  小麦
DOI:
分类号:Q789
基金项目:安徽省教委自然科学基金资助项目(98JL099)
Affinity chromatography materials for glutamate decarboxylase
CHEN Fu xue 1,WEI Qing xin 2,CAO Jun ping 2,ZHENG Xin min 2,GUO Hui 3,CHEN Shi 3,ZHANG Ying han 4,CHEN Fu xue 1,WEI Qing xin 2,CAO Jun ping 2,ZHENG Xin min 2,GUO Hui 3,CHEN Shi 3,ZHANG Ying han 4,CHEN Fu xue 1,WEI Qing xin 2,CAO Jun ping 2,ZHENG Xin min 2,GUO Hui 3,CHEN Shi 3,ZHANG Ying han 4
Abstract:
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 Sepharose with coupled δ-carboxylic group,partly absorbed by other affinity gels.Glutamate could elute the enzyme but inhibit the enzyme activity.Glutamine and 4-aminobytrate could not elute the enzyme at the same concetration.
Key words:  glu sepharose  glutamate decarboxylase  affinity chromatography  wheat

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