摘要
目的:建立稳定性好,制备简单的固定化酶微反应器,旨在为多肽和蛋白质的分析研究提供一种稳定、有效及可行的酶解手段.方法:将前期人工合成类天然酶的环糊精模拟酶β-benzyme通过原位聚合反应制备成新型的毛细管固定化环糊精模拟酶反应器,最后通过对N-苯甲酰-L-精氨酸乙酯(BAEE)作底物的水解效率进行评估.结果:酶反应器的米氏常数和最大酶解速率分别为0.70 mmol/L和2.12 mmol/s,经12周的测试后依然保持稳定.结论:模拟酶反应器活性良好,稳定性好,这为解决蛋白质分析与鉴定研究中的问题提供新技术.
Aim ： In order to provide a stable and effective method for enzymatic hydrolysis of protein, immobilized enzyme microreactor （IMEMR） with simple preparation and good reproducibility was devel-oped. Methods A novel monolithic capillary enzymatic microreactor was first prepared with in situ co-polymerizing specific designed β-cyclodextrin-mimic enzymes β-benzyme. Finally, to confirm the possi-bility of applying the monolithic mimic enzyme reactor for effective protein digestion, the digestion reac-tions performed on column and in solution were evaluated through the hydrolysis of N-benzoyl-L-arginine- ethylester （ BAEE） . Results： The Michael was constant and maximum digestion rate of the mimic en-zyme reactor were 0. 70 mmol/L and 2. 12 mmol/s respectively. The mimic enzyme reactor kept stable af-ter a twelve-week successive test. Conclusion ： The mimic enzyme reactor performed good activity and stability, which opens up a new source of technology in solving protein identification and proteomics char-acterization problems.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2016年第6期454-459,共6页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
国家自然科学基金青年科学基金项目(81503031)
广东省自然科学基金博士科研启动项目(2014A030310506)
广东省科技计划项目(2016A020226004)
广东省医学科研基金项目(A2015351)资助项目
关键词
环糊精
模拟酶
酶反应器
整体柱
cyclodextrins
mimetic enzymes
microreactor
monolithic column
