摘要
采用金属鳌合亲和层析技术,以EIP-ARG-IDA-Co^2＋为载体,从可溶性总蛋白中一步分离纯化具有His-tag标签的GL-7ACA酰化酶.在此基础上进行了固定化方面的研究,其固定化最适条件是150-200U/g给酶量与载体比,固定化时间16 h,固定化介质pH=7.0.固定化酶的最适反应温度为50℃,最适反应pH为8.0.同时还考察了固定化酶的热稳定性、重复利用性及载体的重复使用等特性.
Glutaryl-7-aminocephalosporanic acid acylase（GL-7ACA acylase） is one of important industrial enzymes for the production of 7-aminocephalosporanic acid （7-ACA）, in this paper, a new metal chelating carrier（EIP-ARG-lDA-Co^2＋ ） was prepared to immobilize GL-7ACA acylase. This support can efficiently purify and immobilize GL-7-ACA acylase with His-tag from evaluated proteins in one-step. On the process of immobilization, the optimum immobilization conditions of enzyme were as follows ： enzyme load was 150 - 200 U/g carriers, time was 16 h, pH was 7.0. The optimum temperature and pH of immobilized enzyme was 50 ℃ and 8.0 respectively, it showed that immobilized enzyme was superior to free enzyme at pH and temperature. In addition, Michaelis constant, stabilities to heat, operational stability and carrier reuse were compared and studied. These results indicated that immobilized GL-7-ACA acylase in this method can be applicable for industral production.
出处
《江西师范大学学报（自然科学版）》
CAS
北大核心
2008年第5期549-554,共6页
Journal of Jiangxi Normal University(Natural Science Edition)
基金
江西省科技厅工业攻关项目(00526)
