3种固定化尖吻蝮蛇毒金属蛋白酶的比较  

Preparation and Characterization of Three Immobilized Metalloproteinase from the Venom of Deinagkistrodon acutus

作　　者：聂学奎 和七一[1] 黄达春[1] 陈峻波 余晓东[1] NIE Xuekui;HE Qiyi;HUANG Dachun;CHEN Junbo;YU Xiaodong(Chongqing Engineering Research Center of Bioactive Substance,Ministry of Education Engineering Research Center of Active Substance and Biotechnology,College of Life Science,Chongqing Normal University,Chongqing 401331,China)

机构地区：[1]重庆师范大学生命科学学院教育部活性物质生物技术工程研究中心重庆市生物活性物质工程研究中心,重庆401331

出　　处：《重庆师范大学学报（自然科学版）》2021年第3期107-113,共7页Journal of Chongqing Normal University:Natural Science

基　　金：国家自然科学基金(No.31470570);重庆自然科学基金(No.cstc2017chmsxdny0246);重庆师范大学基金项目(No.17XLB01311,No.19XLB006);中央林业改革发展资金科技推广项目(渝林科推2019-2)。

摘　　要：【目的】筛选出一种最优的固定化尖吻蝮(Deinagkistrodon acutus)蛇毒金属蛋白酶(Snake venom metalloproteinase,SVMP)的方法。【方法】采用D380大孔树脂、海藻酸钠-壳聚糖、溶胶-凝胶等3种载体固定化SVMP,用分光光度法检测固定化金属蛋白酶和游离金属蛋白酶的酶活性,计算它们的酶搭载率、酶活回收率,并检验它们的温度耐受性、pH耐受性和重复使用性,并由此筛选出最优固定化方法。【结果】溶胶-凝胶固定化酶、海藻酸钠-壳聚糖固定化酶和D380大孔树脂固定化酶的酶搭载率分别为91.47%,78.14%和49.44%;D380大孔树脂固定化酶、海藻酸钠-壳聚糖固定化酶和溶胶-凝胶固定化酶的酶活回收率分别为39.00%,39.53%和41.17%;重复使用4次后,溶胶-凝胶固定化酶的酶活回收率最高;溶胶-凝胶固定化酶pH耐受性最优,对温度的耐受性也最高。【结论】溶胶-凝胶固载体固定尖吻蝮蛇SVMP的效果最优。[Purposes]To screen an optimal method for immobilization of snake venom metalloproteinase(SVMP)from Deinagkistrodon acutus.[Methods]The immobilization of SVMP was carried out by using three carriers,including D380 macroporous resin,sodium alginate-chitosan and sol-gel.The enzymatic activities of immobilized and free metalloproteinases were measured spectrophotometrically,their enzyme loading rates and enzyme activity recoveries were calculated,and their temperature tolerance,pH tolerance and reusability were examined,and the optimal immobilization method was screened.[Findings]The enzyme loading rates of sol-gel immobilized enzyme,sodium alginate-chitosan immobilized enzyme and D380 macroporous resin immobilized enzyme were 91.47%,78.14%and 49.44%,respectively;the enzyme activity recovery rates of D380 macroporous resin immobilized enzyme,sodium alginate-chitosan immobilized enzyme and sol-gel immobilized enzyme were 39.00%,39.53%and the enzyme activity recovery of lyso-gel immobilized enzyme was the highest after four times of repeated use;the pH tolerance of lysogel immobilized enzyme was the best and the temperature tolerance was also the highest.[Conclusions]The sol-gel immobilized enzyme had the best effect on the immobilization of SVMP from the sharp-nosed pit viper.

关 键 词：固定化酶 D380大孔树脂载体 海藻酸钠-壳聚糖载体 溶胶-凝胶载体 蛇毒金属蛋白酶(SVMP) 尖吻蝮 

分 类 号：O221.6[理学—运筹学与控制论]