鳗弧菌胞外金属蛋白酶的化学修饰研究  

Chemical Modification of the Metallopretease Isolated from Pathogenic Vibrio Anguillarum

作　　者：池政豪[1] 陈吉祥[1] 杨慧[1] 刘斌[1] 李筠[1] 

机构地区：[1]中国海洋大学海洋生命学院,青岛266003

出　　处：《生物技术通报》2006年第C00期434-437,444,共5页Biotechnology Bulletin

基　　金：国家863计划项目(2003AA622070);国家自然科学基金项目(30371108)

摘　　要：用DEPC、EDC、DTNB、PMSF等8种化学修饰剂对鳗弧菌胞外金属蛋白酶进行了化学修饰。结果表明化学修饰后酶的活力发生了改变,其中组氨酸、酸性氨基酸、半胱氨酸残基的化学修饰引起酶活性的明显降低,说明组氨酸残基、酸性氨基酸、半胱氨酸残基及其二硫键在维持酶活力中发挥重要作用,是酶活力所必需;而对精氨酸、丝氨酸、ε-氨基等修饰后酶活性影响较小,表明不是酶的活性所必须的基团。The chemical modification of metalloprotease isolated from Vibrio anguillarum was studied by eight protein modification reagents in order to identify the catalytically essential amino acid residues of the enzyme. The enzyme activity of the modified enzyme changed differently. Modification of histidine residues, carboxyl groups of acidic amino acid residues, disulfides groups of cystine residues by diethylpyrocarbonate （DEPC） , carbodiimide （EDC） , 5,5 dithio-bis-（-2-nitrobenzoic acid） （DTNB）, 2-Mecarptoethonal （2-ME） and dithiothreitol （DTT） decreased the enzyme activity greatly, which indicated that these amino acid residues were essential to catalytic activity of the enzyme. Modification of guanidine groups of arginine residues, serine residues, e-amino groups by acetylacetone, phenylmethylsulphonylfluoride （PMSF） , formaldehyde had no effects on the enzyme activity, which suggested that these amino acid residues were not essential to activity of the metalloprotease.

关 键 词：鳗孤菌 胞外蛋白酶 化学修饰 酶活性 

分 类 号：S941[农业科学—水产养殖]