Isolation and physicochemical characterization of polysaccharide fractions isolated from Schisandra chinensis
Isolation and physicochemical characterization of polysaccharide fractions isolated from Schisandra chinensis
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DOI：
10.1007/s10600-012-0116-5
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发表时间：
2012-01
影响因子：
0.8
通讯作者：
Haibin Tong;Bing Zhao;Fengguo Du;D. Tian;K. Feng;Xin Sun
中科院分区：
文献类型：
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作者：
Haibin Tong;Bing Zhao;Fengguo Du;D. Tian;K. Feng;Xin Sun
在过去的几十年里，人们认为植物中的多糖及其结合物不仅是能源，而且在许多生命过程中发挥着至关重要的生物学作用[1]。生物活性多糖对多种疾病的药效结构和机制已被广泛研究，许多具有不同疗效的天然多糖已被研究甚至应用于治疗中[2]。五味子广泛分布于中国东北地区，作为中药使用已有数千年的历史[3]。已正式收载于《中国药典》，索引为滋补镇静药。 1596年（公元前2697年）《神农本草经》中也将其列为止咳、平喘的上品[4, 5]。令人意外的是，目前还没有针对五味子多糖的具体研究。因此，本研究对五味子多糖组分进行分离纯化，并进一步研究其基本理化性质，以期最有效地获得高性能多糖产品，挖掘五味子的应用潜力。本研究采用AKTA纯化系统，采用Hiload 16/30纯化五味子多糖组分。 DEAE-纤维素柱和 Hiload 26/100 Sephacryl S-200 柱。从五味子中提取的粗水溶性多糖的得率为干燥物的7.8％。经过冻融过程以及蛋白酶和 Sevag 方法组合的脱蛋白后，将粗多糖样品 (cSCPS) 加载到 DEAE-纤维素柱上，并用去离子水和 0o1 M 梯度的 NaCl 溶液以 1 mL/min 的流速进行洗脱。收集去离子水洗脱的主要部分，冻干，并进一步在Sephacryl S-200柱上分级，并用0.15M NaCl溶液以0.5mL/min的流速洗脱。分离三个主要组分（SCPS-a、SCPS-b 和 SCPS-c）以进一步分析物理化学性质。多糖级分的总糖、蛋白质、糖醛酸含量、分子量和单糖组成总结于表1中。多糖级分SCPS-b和SCPS-c的总碳水化合物含量（分别为93.6%和95.8%）高于SCPS-a（79.3%）。根据Bradford法测得SCPS-a的蛋白质含量为19.6%；在 SCPS-b 和 SCPS-c 中未检测到它。
In the past decades, it was believed that polysaccharides and their conjugates in plant are not only energy resources but also play crucial biological roles in many life processes [1]. The structure and mechanisms of pharmaceutical effects of bioactive polysaccharides on various diseases have been extensively studied, and many natural polysaccharides with different curative effects have been examined and even applied in therapies [2]. Schisandra chinensis, distributed abundantly in the northeast region of China, has been used in traditional Chinese medicine for thousands of years [3]. It is officially listed in the Chinese Pharmacopoeia and indexed as a tonic and sedative. It is also listed in the “Shen Nong Ben Cao Jing” book, year 1596 (2697 BC) as a superior drug that helps in coughs and prevents asthma [4, 5]. Unexpectedly, at present there are no specific studies on polysaccharides from S. chinensis. Therefore, the present studies were carried out to isolate and purify the polysaccharide fractions from S. chinensis and to further investigate their basic physicochemical properties in order to most effectively acquire high-performance polysaccharide products and exploit the applied potential of S. chinensis.In this study, the AKTA purification system was employed to purify S. chinensis polysaccharide fractions with Hiload 16/30 DEAE-cellulose column and Hiload 26/100 Sephacryl S-200 column. The yield of the crude water-soluble polysaccharide extracted from S. chinensis was 7.8% of dried material. After the freeze–thaw process and deproteination by a combination of proteinase and the Sevag method, the crude polysaccharide sample (cSCPS) was loaded onto the DEAE-cellulose column and eluted with de-ionized water and 0o1 M gradient of NaCl solution at a flow rate of 1 mL/min. The main fraction eluted by de-ionized water was collected, lyophilized, and further fractionated onto a Sephacryl S-200 column and eluted with 0.15 M NaCl solution at a flow rate of 0.5 mL/min. Three main fractions (SCPS-a, SCPS-b, and SCPS-c) were separated for further analysis of physicochemical properties. The total sugar, protein, uronic acid contents, molecular weight, and monosaccharide compositions of the polysaccharide fractions are summarized in Table 1. The polysaccharide fractions SCPS-b and SCPS-c had a higher total carbohydrate content (93.6% and 95.8%, respectively) than SCPS-a (79.3%). According to the Bradford method, the protein content of SCPS-a was 19.6%; it was not detected in SCPS-b and SCPS-c.