以伴刀豆球蛋白为介质定向固定化脲酶的研究.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2008, April 25; 24(4): 617-621 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn © 2008 Institute of Microbiology, CAS CSM, All rights reserved 研究报告 周建芹, 陈韶华, 王剑文 , 215123 : 将戊二醛将伴刀豆球蛋白(ConA)和壳聚糖载体交联, 然后利用ConA 与脲酶糖链的特异性结合作用, 实现脲酶 的定向固定化。定向固定化的最适条件为戊二醛浓度3.5%、ConA 浓度1 mg/mL、ConA 溶液pH 值7.0 、脲酶浓度 0.4 mg/mL 。 定向固定化脲酶的最适pH 5.0~6.0 、最适温度77°C 、米氏常数Km11.76 mmol/L, 与游离酶及非定向固定化脲酶比较, 定 向固定化脲酶的最适pH 向酸性范围发生了偏移并有更宽的pH 适用范围, 最适温度提高, 与底物的亲和力较大, 且有较 好的操作稳定性。 : 脲酶, 伴刀豆球蛋白, 定向固定化 Research on the Orientedly Immobilized Urease via Concanavalin A Jianqin Zhou, Shaohua Chen, and Jianwen Wang School of Pharmacy, Soochow University, Suzhou, 215123, China Abstract: Concanavalin A (ConA) is immobilized on a pre-activated chitosan microspheres, and then oriented immobilization of urease is carried out based on the strong interaction between ConA and glycoprotein. The optimum immobilization conditions are as follows: glutaraldehyde concentration is 3.5%, ConA concentration 1mg/mL, ConA pH 7.0 and urease concentration 0.4 mg/mL. For orientedly immobilized urease, the highest activity was allowed at pH 5.0~6.0 and temperature 77°C, and the Michaelis constant (Km) was disclosed to be 11.76 mmol/L by Lineweaver-Burk plot. Compared with the free urease and the randomly immobilized urease, the optimum pH of the orientedly immobilized urease becomes smaller and the pH domain wider. Orientedly immobilized urease presents higher temperature resistance, higher affinity to the substrate, and higher stability of operation. Keywords: urease, concanavalin A, oriented immobilization ,