β-葡萄苷酶转化松脂醇二葡萄糖苷元的制备工艺  

Optimizing the Preparation Process ofβ-Glucosidase Transforming PDG-A by Response Surface Methodology

作　　者：刘洋[1] 付冬梅 张良[3] 张琳[2] 祖元刚[2] Liu Yang(College of Forestry,Northeast Forestry University,Harbin Heilongjiang 150040)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040 [2]东北林业大学化学化工与资源利用学院森林植物生态学教育部重点实验室林业生物制剂教育部工程研究中心,黑龙江哈尔滨150040 [3]黑龙江中医药大学附属第二医院南院,黑龙江哈尔滨150060

出　　处：《黑龙江医药》2021年第1期5-9,共5页Heilongjiang Medicine journal

基　　金：中央高校基本科研业务费专项资金(编号:2572015CA06);林业公益性行业科研专项(编号:201504701-205)。

摘　　要：目的:利用β-葡萄糖苷酶催化水解杜仲主要降压活性成分松脂醇二葡萄糖苷(PDG),制备其苷元(PDG-A)。方法:对水解反应时间、p H及温度等条件进行优化,并采用液相质谱联用和核磁共振波谱等手段对产物进行结构解析。结果:β-葡萄糖苷酶水解PDG的最佳条件为:水解时间6h,水解p H5,水解温度50℃,三次实验的PDG-A收率分别为98.52%、97.91%、98.69%;PDG的水解产物为PDG(M=682)水解掉2个葡萄糖基团(M=162)而成的苷元结构(M=358),结合1HNMR及13C-NMR确定其为苷元PDG-A。结论:β-葡萄糖苷酶催化水解PDG制备其苷元PDG-A的反应温和、转化率98%以上,此方法可以为大规模制备PDG-A提供理论指导。Objective:Usingβ-glucosidase to prepare pinoresinol diglucoside aglycone(PDG-a)from pinoresinol diglucoside(PDG),an antihypertensive active components of Eucommia ulmoides Oliv.Methods:The reaction time,p H and temperature of the hydrolysis were optimized,and the structure of the product was analyzed by liquid mass spectrometry and NMR.Results:the optimal conditions of PDG hydrolysis byβ-glucosidase were as follows:hydrolysis time 6.17 h,hydrolysis p H 5.03,hydrolysis temperature 49℃.The hydrolysis rates of the three validation experiments were 98.52%,97.91%and 98.69%,respectively,which were consistent with the predicted value 98.72%.PDG(M=682)hydrolyzed two glucose groups(M=162)to form aglycone structure(M=358),which was identified as the aglycone of PDG by1 H-NMR and13 C-NMR.Conclusion:β-glucosidase can catalyze the hydrolysis of PDG into PDG-A with mild reaction and over 98%conversion rate.This method can provide theoretical guidance for large-scale preparation of PDG-A.

关 键 词：松脂醇二葡萄糖苷 Β-葡萄糖苷酶 松脂醇二葡萄糖苷元 响应面法 

分 类 号：R284.2[医药卫生—中药学]