茯苓药渣酶解工艺及其产物初步分析  

Optimization of Enzymatic Preparation Process from the Residue of Poria cocos(Schw.) Wolf and its Products Analysis

作　　者：张年 匡创富[3] 段彦 曾弦 戴甲木 李娟 李顺祥 ZHANG Nian;KUANG Chuang-fu;DUAN Yan;ZENG Xian;DAI Jia-mu;LI Juan;LI Shun-xiang(First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan Province Engineering Research Center of Bioactive Substance Discovery of Chinese Medicine,School of Pharmacy,Hunan University of Chinese Medicine,Changsha 410208,China;Second Hospital of Hunan University of Chinese Medicine,Changsha 410005,China;Hunan Academy of Fuling Research,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖长沙410007 [2]湖南中医药大学药学院,湖南省中药活性物质筛选工程技术研究中心,湖南长沙410208 [3]湖南中医药大学第二附属医院,湖南长沙410005 [4]湖南补天茯苓研究院,湖南长沙410208

出　　处：《时珍国医国药》2022年第10期2501-2507,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家科技部“十二五”国家科技支撑计划项目(2011BAI01B09);湖南省长沙市科学技术局国际合作重点项目(kq1907135);湖南中医药大学中药学一流建设学科(校行科字[2018]3号)。

摘　　要：目的优化茯苓药渣酶解工艺,制备水溶性酶解产物,并初步分析酶解产物的结构。方法通过Box-Behnken设计-响应面法优化制备工艺,酶解产物经Sevage法脱蛋白,Sephadex G-75、DEAE Sepharose F F以及Sephadex G-50纯化,采用PMP柱前衍生-高效液相色谱法(HPLC)和高效凝胶渗透色谱-多角度激光光散射-示差折光联用(HPGPC-MALLS-RI)测定其单糖组成和分子量,刚果红实验检测三股螺旋结构的有无。结果使用蜗牛酶,反应温度49℃,pH 6.2,反应时间34 h,酶用量2.6%,酶解得率达到36.97%左右,纯化后产物主要是由葡萄糖组成的低聚糖,由3个组分构成,重均分子量分别为1.739×10^(5)、811.9、435.4,其中重均分子量为811.9的低聚糖为中性糖,占92.1%,是主要组成部分,纯化酶解产物不具有三股螺旋结构。结论蜗牛酶适用于降解茯苓药渣,工艺稳定可行,所得产物得率较高,水溶性好,为进一步开发利用茯苓资源奠定了基础。Objective To optimize the preparation process of water-soluble polysaccharides or oligosaccharide from the residue of Poria cocos(Schw.)Wolf and to preliminarily analyze the structure of enzymolysis products.Methods Box-Behnken design-response surface method was used to study the optimum preparation conditions.The protein was deproteinized by Sevage method,and the enzymolysis products was purified by Sephadex G-75,DEAE Sepharose F F and Sephadex G-50 column chromatography.The monosaccharide composition and molecular weight was determined by HPLC with PMP precolumn derivatization and HPGPC-MALLS-RI.The Congo red assay was used to detect the existence of three-stranded helical structures.Results Using snailase,with the optimum hydrolysis condition for Poria cocos enzymatic hydrolysis of 49℃,pH 6.2,34 h,the enzyme dosage was 2.6%,the extraction rate of pachyman was about 36.97%.After purified,polysaccharide were main composed of glucose was made up of three components,which Mw was 1.739×10^(5),811.9,and 435.4,and the oligosaccharide of the main component having Mw of 811.9 accounted for 92.1%.There was no triple helix structure existed in this fraction.Conclusion The processing technology of selecting Snailase and hydrolyzing polysaccharides optimized by Box-Behnken design-response surface method has the advantages of stable and a new processing technology for water solubility enzymolysis products.which provides some ideas for the further development and utilization of poria cocos resources.

关 键 词：茯苓 药渣 酶水解 产物分析 

分 类 号：S567[农业科学—中草药栽培]