基于高分辨质谱技术的黄芪多糖APS-Ⅱ酶解活性寡糖特征分支位点的研究
Study on the characteristic branch sites of oligosaccharides of Astragalus polysaccharide APS-Ⅱ enzymolysis based on high resolution mass spectrometry
刘宇冲 1李虎峰 1李科 2秦雪梅 1杜昱光 3李震宇1
作者信息
- 1. 山西大学中医药现代研究中心,山西太原 030006;山西大学化学,生物学与分子工程教育部重点实验室,山西太原 030006;地产中药功效物质研究与利用山西省重点实验室,山西太原 030006
- 2. 山西大学中医药现代研究中心,山西太原 030006;山西大学化学,生物学与分子工程教育部重点实验室,山西太原 030006;地产中药功效物质研究与利用山西省重点实验室,山西太原 030006;中国科学院过程工程研究所,北京 100190
- 3. 中国科学院过程工程研究所,北京 100190
- 折叠
摘要
黄芪多糖是黄芪中免疫调节活性最强,含量最为丰富的物质,具有抗肿瘤、抗病毒、免疫促进等多种生物活性,临床应用广泛.前期研究发现黄芪多糖主要由2种不同相对分子质量多糖APS-I(＞2 000 kDa)、APS-Ⅱ(10 kDa)组成,APS-Ⅱ(10 kDa)为黄芪多糖中活性最强组分,并采用α-1,4-葡聚糖内切酶将APS-Ⅱ降解为寡糖,通过体外免疫活性筛选发现2～9糖整体免疫活性较低,而10～14糖具有较强的免疫活性,且活性优于未降解的APS-Ⅱ.为了探究APS-Ⅱ降解寡糖发挥免疫活性的关键结构,本文采用MALDI-TOF-MS生物质谱以及高分辨质谱仪器ESI-Q Exactive-MS对APS酶解寡糖进行解析,通过对比发现10～14糖中存在1→4和1→6两种连接方式共存的支链结构,推测1→4和1→6两种连接方式共存的支链结构为APS-Ⅱ发挥免疫活性的关键结构,为黄芪多糖和寡糖的构效关系研究奠定理论基础.
Abstract
Astragalus polysaccharides are the most immunoregulatory active and abundant substances in Astragalus,with anti-tumor,anti-viral,and immune-promoting biological activities.They have been widely used in clinical practice.Previous studies have found that Astragalus polysaccharides are mainly composed of two different polysaccharides,APS-Ⅰ(＞2 000 kDa)and APS-Ⅱ(10 kDa),with APS-Ⅱ(10 kDa)being the most active component of Astragalus polysaccharides.We used α-1,4-glucan endo-hydrolysis enzyme to degrade APS-Ⅱ into oligosaccharides,and screened the immune activity of oligosaccharides in vitro.We found that the overall immune activity of 2-9 oligosaccharides was low,while the immune activity of 10-14 oligosaccharides was strong,and the activity was better than that of untreated APS-Ⅱ.To investigate the key structural features of APS-Ⅱ oligosac-charides that play a role in immune activity,we used MALDI-TOF-MS biochemical mass spectrometry and high-resolution mass spectrometry instrument ESI-Q Exactive-MS to analyze the APS enzymatic oligosaccharides.By comparing,we found that 10-14 oligosaccharides contain 1→4 and 1→6 branched structures with coexisting linkages,suggesting that 1→4 and 1→6 linkages in branched structures are key structural features that play a role in the immune activity of APS-Ⅱ,laying a theoretical foundation for the structure-activity relationship of Astragalus polysaccharides and oligosaccharides.
关键词
APS-Ⅱ/酶解/连接方式/结构解析/高分辨质谱Key words
APS-Ⅱ/enzymolysis/connection mode/structural analysis/high resolution mass spectrometry引用本文复制引用
基金项目
国家自然科学基金资助项目(81872962)
国家博士后科学基金资助项目(2019M650851)
国家重点研发计划(2019YFC1710800)
山西省重点研发计划重点项目(201603D311101)
山西省优秀人才科技创新项目(201605D211030)
山西省优秀人才科技创新项目(201705D211020)
山西省科技创新人才团队专项基金()
出版年
2024
NETL
NSTL
万方数据