金针菇乙酰胆碱酯酶抑制剂筛选条件的优化与应用  

Optimization and application of acetylcholinesterase inhibitor screening criteria from the fruiting body of Flammulina velutipes

作　　者：余淑英[1] 卢艳[1] 王星丽 瞿亮 孙培龙[1] 张安强[1] Yu Shuying Lu Yan Wang Xingli Qu Liang Sun Peilong Zhang Anqiang(Department of Food Science and Technology, Zhejiang University of Technology, Hangzhou 310014, China Hangzhou Baishanzu Biological Science and Technology Co. Ltd, Hangzhou 310052, China)

机构地区：[1]浙江工业大学食品科学技术系,浙江杭州310014 [2]杭州百山祖生物科技有限公司,浙江杭州310052

出　　处：《食药用菌》2017年第1期34-39,共6页Edible and Medicinal Mushrooms

基　　金：国家十二五科技支撑项目"食用菌深加工产品开发与产业化"(2013BAD16B07)

摘　　要：经优化乙酰胆碱酯酶抑制剂的筛选条件,结合体外乙酰胆碱酯酶的活性,筛选出具有抗阿尔茨海默的高抑制活性的均一金针菇子实体多糖组分;采用改良的Ellman法,建立乙酰胆碱酯酶抑制剂的筛选系统,考察其最大吸收波长、ACh E的浓度、ATCI的浓度、显色剂DTNB的浓度,以及反应体系的时间对系统的影响,最终确定反应体系的最佳条件为:酶反应体系的反应温度37℃,反应时间30 min,最佳检测波长412 nm,ACh E的浓度0.05 u/m L,ATCI的浓度0.10 mmol/L,p H=8.0的100 mmol/L PBS缓冲溶液,显色剂DTNB终浓度0.066 7mmol/L,通过乙酰胆碱酯酶抑制剂的筛选发现了金针菇多糖抑制剂。In order to optimize the screening conditions for acetylcholinesterase inhibitors and combine with in vitro activity of acetylcholinesterase and screen with high inhibiting activity of anti-Alzheimer uniform Flammulina polysaccharide.The improved Ellman method was used to establish an acetylcholinesterase inhibitor screening system and investigate influences on the system by the maximum absorption wavelength,the concentration of ACh E,ATCI concentration,the concentration of reagent DTNB,time of system,and ultimately determine the optimum conditions of the reaction system.A screening model for ACh EIs was optimized with temperature 37 ℃,reaction time 30 min,wavelength 412 nm,the concentration of ACh E 0.05 u/m L,a concentration of ATCI 0.10 mmol/L,100 mmol/L PBS buffer solution p H=8.0,color agent DTNB final concentration 0.066 7 mmol/L;Through the screening of acetylcholinesterase inhibitors found FVP inhibitors.

关 键 词：金针菇多糖 乙酰胆碱酯酶抑制剂 筛选条件优化 

分 类 号：R91[医药卫生—药学]