杂色鲍组织蛋白酶L的分离纯化与性质研究  

Purification and characterization of cathepsin L from abalone(Haliotis Diversicolor)

作　　者：董训江[1] 钟婵[1] 蔡秋凤[1] 曹敏杰[1] 

机构地区：[1]集美大学生物工程学院,福建厦门361021

出　　处：《食品工业科技》2011年第12期247-250,309,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金(31071519);福建省自然科学基金(2011J01227)

摘　　要：通过硫酸铵分级沉淀、SP-Sepharose阳离子交换层析、Sephacryl S-200HR凝胶过滤和Hydroxyapatite羟基磷灰石层析,从杂色鲍(Haliotis diversicolor)内脏中分离纯化了组织蛋白酶L。SDS-PAGE结果显示,纯化蛋白的分子量约为28ku。该酶最适温度37℃,最适pH5.5。当温度低于40℃,pH在5.0～6.5范围内该酶较稳定。底物特异性实验与抑制剂实验结果表明,该酶属于半胱氨酸蛋白酶。金属离子Mn2+、Ba2+和Mg2+对该酶有不同程度的激活作用,而Co2+、Cu2+、Zn2+、Fe2+和Ca2+等对该酶起抑制作用。Cathepsin L was purified from the hepatopancreas of abalone（Haliotis diversicolor）by ammonium sulfate fractionation,and chromatographies including SP-Sepharose,Sephacryl S-200 gel filtration and hydroxyapatite prepacked column. The molecular mass of the purified enzyme was estimated to be 28ku by SDS-PAGE. Optimum temperature and pH of the purified enzyme were 37℃ and 5. 5,respectively. The enzyme was stable in the range of pH from 5. 0 to 6. 5 and at temperature lower than 40℃. Substrate specificity and inhibitor sensitivity analysis revealed that the enzyme was a cysteine proteinase. A survey of effects of metal ions on the enzyme showed that metal ions including Mn2＋,Ba2＋ and Mg2＋ activated the enzyme to some degree while Co2＋,Cu2＋,Zn2＋,Fe2＋and Ca2＋ revealed inhibitory effect.

关 键 词：杂色鲍 组织蛋白酶L 分离纯化 性质研究 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]