首页|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

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

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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。
Study on the characteristic branch sites of oligosaccharides of Astragalus polysaccharide APS-Ⅱ enzymolysis based on high resolution mass spectrometry
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.

APS-Ⅱenzymolysisconnection modestructural analysishigh resolution mass spectrometry

刘宇冲、李虎峰、李科、秦雪梅、杜昱光、李震宇

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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

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APS-Ⅱ 酶解 连接方式 结构解析 高分辨质谱

国家自然科学基金资助项目国家博士后科学基金资助项目国家重点研发计划山西省重点研发计划重点项目山西省优秀人才科技创新项目山西省优秀人才科技创新项目山西省科技创新人才团队专项基金

818729622019M6508512019YFC1710800201603D311101201605D211030201705D211020

2024

10.16438/j.0513-4870.2023-1379

药学学报
中国药学会 中国医学科学院药物研究所

药学学报

CSTPCD北大核心
影响因子：1.274
ISSN：0513-4870
年,卷(期)：2024.59(7)
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