以卵清蛋白为载体的固定化木瓜蛋白酶的制备及其性质  

Preparation and its properties of the immobilized papain with denatured egg white protein as the support

作　　者：黄宜兵[1] 吴晓霞[1] 吴胜男[1] 徐力[1] 张学忠[1] 

机构地区：[1]吉林大学分子酶学工程教育部重点实验室,长春130023

出　　处：《吉林大学学报（理学版）》2004年第4期616-621,共6页Journal of Jilin University:Science Edition

基　　金："十五"国家科技攻关计划项目基金(批准号:2001BA960C).

摘　　要：以变性的卵清蛋白为载体,用戊二醛为交联剂固定木瓜蛋白酶.通过正交实验考察了固定化pH值、交联剂戊二醛浓度、交联时间和酶用量对固定化木瓜蛋白酶活力的影响,确定了最适固定化条件:5%戊二醛,pH6.0,交联时间20h,每克载体含酶量12mg.对比研究了游离木瓜蛋白酶和固定化木瓜蛋白酶的性质,所得到的固定化木瓜蛋白酶最适温度90℃,最适pH9.0,米氏常数53.6mg/mL,其热稳定性及耐热性均较游离酶显著提高.Papain was immobilized on a denatured egg white protein as the support with glutaraldehyde as the cross-linker. The effects of pH, the concentration of glutaraldehyde, time and the amount of the enzyme on the activity of the immobilized papain were examined through the experiments of orthogonal test. The optimal condition for the immobilization was set up, i.e., glutaraldehyde concentration is 5%; pH 6.0; the quantity of enzyme used 12 mg/(g support) and the reaction time 20 h. At the same time, the comparative study on the properties of free and immobilized papain was made. The optimal temperature and pH are 90 ℃ and 9.0, (respectively.) The (K_m) for the immobilized enzyme is (53.6) mg/mL. The thermostability and thermotolerance of the immobilized enzyme are obviously increased compared to those of the free one.

关 键 词：戊二醛 卵清蛋白 载体 游离 对比研究 木瓜蛋白酶 固定化 交联时间 交联剂 制备 

分 类 号：R392[医药卫生—免疫学] TQ325.12[医药卫生—基础医学]