壳聚糖固定化谷胱甘肽硫转移酶的研究  

Study on the Immobilization of Glutathione S-transferases on Chitosan

作　　者：尹登科[1] 丁虹[1] 喻昕[1] 

机构地区：[1]武汉大学药学院生物技术药学系,湖北武汉430072

出　　处：《生物技术》2004年第6期17-19,共3页Biotechnology

摘　　要：目的 :利用壳聚糖固定日本血吸虫谷胱甘肽硫转移酶 ,并对固定化酶性质及体外催化活性进行研究。方法 :利用大肠杆菌BL2 1(DE3)表达日本血吸虫谷胱甘肽硫转移酶 ,并从中粗提谷胱甘肽硫转移酶 ,将其固定在用戊二醛交联的壳聚糖载体上 ,对游离酶和固定酶的最适pH、温度 ,游离酶和固定化酶对底物 1-氯 - 2 ,4 -二硝基苯 (CDNB)和谷胱甘肽 (GSH)的亲和力 ,温度的稳定性进行了研究。结果 :固定化酶活回收率可达 4 1.6 % ,最适pH6 .5～ 7.0 ,最适温度 37℃ ,对底物 (CDNB和GSH)的亲和力略有下降 ,对温度稳定性大大提高。Objective:Using Chitosan to Immobilize the Glutathione S-transferases(GST) of Schistosoma japonicum and studying the character of the immobilized GST and the catalyze ability in vitro.Methods:Using E.coli BL21(DE3) to express Glutathione S-transferase(GST) of Schistosoma japonicum,and the crude extract of GST was immobilized on Chitosan and cross-linked with gluaraldehyde.Optimal pH,temperature,affinity to substrate 1-chloro-2,4-dinnitrobenzen(CDNB) and glutathione(GSH) and thermal stability of immobilized GST were investigated and compared with the free enzyme.Results:The activity recovery of immobilized GST reached 41.6%,optimal pH 6.5～7.0,optimal temperature 37℃ and the thermal stability of immobilized GST was enhanced.Immobilized GST showed high efficacy for detoxification in vitro.

关 键 词：谷胱甘肽硫转移酶 体外 日本血吸虫 外固定 GSH 游离 壳聚糖 最适PH 固定化酶 最适温度 

分 类 号：Q814[生物学—生物工程] R735[医药卫生—肿瘤]