小分子封闭提高固定化嗜热菌蛋白酶的热稳定性  

Enhancement of thermal stability of immobilized thermolysin using end-group blocking

作　　者：王芬[1,2] 陈飞飞[1,2] 高为芳[2] 杜方川[1,3] 王安明[1] 谢恬[2] 

机构地区：[1]杭州师范大学材料与化学化工学院,杭州311121 [2]杭州师范大学生物医药与健康研究中心,杭州311121 [3]杭州师范大学生命与环境科学学院,杭州311121

出　　处：《生物加工过程》2013年第6期47-52,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金(20906016;21076053);教育部长江学者和创新团队发展计划(IRT1231);浙江省自然科学基金(Y13B060058);杭州市农业科研攻关专项(20120232B13)

摘　　要：为了提高固定化嗜热菌蛋白酶的热稳定性,在制备共价固定化嗜热菌蛋白酶的基础上,通过选择氨基酸和醇类小分子来封闭载体表面未反应的活化基团,并考察了固定化酶的催化活性及热稳定性。结果发现:L-Trp和L-Val封闭修饰固定化酶时,在80℃的水浴中加热150 min后其剩余活力仍为93.4%和98.6%,其效果约为未经小分子封闭的固定化嗜热菌蛋白酶的2倍。所筛选的几种小分子物质中,叔戊醇、L-Trp、L-Val及L-Ala不仅能提高固定化嗜热菌蛋白酶的热稳定性,而且也可以提高固定化酶的相对活力,从而更有利于其在工业生产中的应用。In order to enhance the thermal stability of the immobilized thermolysin, unreacted active end groups on the carrier surface were blocked with reagents such as amino acids and alcohols, after the thermolysin was covalently immobilized on the carrier. Both catalytic activity and thermal stability of the immobilized enzyme were investigated. The results showed that after heated at 80 ℃ in water for 150 min, the residual activities of the immobilized enzymes blocked and modified by L-Trp and L-Val were about 93.4% and 98.6%, respectively, which was approximately 2 times higher than that of the immobilized thermolysin. Among the blocking reagents, tert-amyl alcohol, L-Trp, L-Val and L-Ala not only improved the thermal stability of immobilized therrnolysin, but also enhanced the relative activity of the immobilized enzyme, thus it was more useful for its industrial application.

关 键 词：封闭 末端基团 固定化 热稳定性 嗜热菌蛋白酶 

分 类 号：Q814.2[生物学—生物工程]