舟山眼镜蛇毒L-氨基酸氧化酶的分离纯化、理化及酶学性质  

Purification and Properties of the L-amino Acid Oxidase from Chinese Cobra(Naja atra) Venom

作　　者：林丽珊[1] 张志强[2] 陈洲[2] 许云禄[2] 

机构地区：[1]莆田学院药学系,莆田351100 [2]福建医科大学药学院蛇毒研究所,福州350004

出　　处：《福建医科大学学报》2007年第4期308-311,共4页Journal of Fujian Medical University

基　　金：福建省科技厅资助省属高校项目(2006F5049);莆田市科技计划项目(2005S07)

摘　　要：目的从舟山眼镜蛇蛇毒中分离L-氨基酸氧化酶(LAO),测定其理化和酶学性质。方法应用Sephadex G-100凝胶色谱和POROS CM20离子交换色谱分离纯化LAO;Wellner和Lichtenberg法测定LAO活力;SDS-PAGE法测定相对分子质量。结果舟山眼镜蛇蛇毒经Sephadex G-100凝胶色谱和两次POROS 20离子交换色谱后,获得LAO纯品(暂定名NA-LAO)。NA-LAO在非还原和还原条件下相对分子质量均为58 kD左右;其反应最适pH为8.0,最适温度为60℃,对L-苯丙氨酸的米氏常数(Km)为3.08 mmoL/L。结论应用凝胶过滤色谱和离子交换色谱可从舟山眼镜蛇毒中分离纯化LAO。Objective To purify an L-amino acid oxidase（LAO） from the venom of Naja atra. Methods The L-amino acid oxidase was purified from the venom of Naja atra by Sephadex G-100 and ROROS chromatography. Activity of LAO was determined by Wellner and Lichtenberg methods. The molecular weight of this enzyme was tested by SDS-polyacrylamide. Results The LAO designated sa NA-LAO was purified as homogeneity by the chromatography. The molecular weight of NA-LAO was 58 kD under non-reducing or reducing conditions, implied that the enzyme was composed of single chain. The optimum pH for it＇s activity was 8.0 and the optimum temperature was 60 ℃, the Km of L-phenylalanine was 3.08 mmoL/L. Conclusion The NA-LAO can be isolated and purified from the venom of Naja atra. NA-LAO composed with 58 kD molecular weight and characteristic enzyme activity.

关 键 词：眼镜蛇毒液类 氨基酸氧化还原酶 色谱法 凝胶 色谱法 离子交换 浙江 

分 类 号：R99[医药卫生—毒理学]