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Preparation of Immobilized Thrombin and Its Application in Screening and Analyzing Bioactive Compounds from Flos Sophorae Immaturus
Authors
侯梅芳
廖循
+3 more
张波
白小
胡瑾洁
Publication date
1 January 2018
Publisher
'Korean Institute of Science and Technology Information (KISTI)'
Doi
Cite
Abstract
槐米是药食两用植物,有清肝明目和凉血止血之功效;其主要化学成分为黄酮类物质,具有抗氧化、抗凝血、降血压等多种生物活性。本文将凝血酶(THR)固定于氨基修饰的磁珠表面,以其为亲和固相萃取吸附剂,开发出一种快速筛选中药中凝血酶抑制剂的方法。首先合成了表面修饰氨基的磁珠,然后以戊二醛作为交联剂将凝血酶共价固定于磁珠表面得到磁珠固定化的凝血酶(MNP@THR)。利用TEM,VSM,FT-IR等对合成的固定化酶进行了表征,同时获得了优化的合成条件为: THR浓度200 mug/mg, pH值7.0,时间60 min,温度37 ℃为最佳合成条件,此时酶的固定化量为65 mug/mg。利用MNP@THR从槐米提取物中特异性地吸附得到两个活性化合物,并通过磁分离进行前处理后,由HPLC-ESI-MS鉴定其分别为槲皮素和芦丁。通过体外酶活抑制实验测出,槲皮素具有较好的抑制活性,其半数抑制浓度IC_(50)为16.6 0.68 muM;芦丁的凝血酶抑制活性较弱,IC_(50)值为1.5 0.14 mM
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Last time updated on 28/10/2019