黑豆糖蛋白的结构分析及抗氧化和免疫活性  

Structural Characterization and Antioxidant and Immunoregulatory Activity of Glycoproteins from Black Soybeans

作　　者：胡懿化 王星滟 张武霞 岳爱琴[2] 杜维俊[2] 赵晋忠[1] 李鹏[1] HU Yihua;WANG Xingyan;ZHANG Wuxia;YUE Aiqin;DU Weijun;ZHAO Jinzhong;LI Peng(Basic Department,Shanxi Agricultural University,Taigu 030801,China;College of Agriculture,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学基础部,山西太谷030801 [2]山西农业大学农学院,山西太谷030801

出　　处：《食品科学》2021年第17期19-26,共8页Food Science

基　　金：国家自然科学基金青年科学基金项目(31800678,81703945);山西省高校创新基金项目(2019L0365,2019L0360);山西省研究生教育创新项目(2020SY198);山西省重点研发计划项目(201903D211006);山西功能农业共性关键技术研究与示范项目(201703D211001)。

摘　　要：本实验从黑豆中分离得到两种糖蛋白,并对其结构组成、抗氧化活性和免疫活性进行研究,为糖蛋白的高效开发利用提供理论依据。采用碱提醇沉法提取黑豆糖蛋白粗品,命名为HDJ,通过DEAE-52纤维素柱层析和Sephadex G-100柱层析分离纯化获得两个均一组分,分别命名为HDJS2和HDJS5-2。利用物理化学方法和光谱法对其化学组成及结构进行分析,通过测定总还原力、1,1-二苯基-2-三硝基苯肼(2,2-diphenyl-1-picrylhydrazyl,DPPH)清除率及总抗氧化能力研究两种糖蛋白的抗氧化活性,采用噻唑蓝比色法评价两种糖蛋白的免疫活性。结果表明:HDJS2(3.20×104 Da)和HDJS5-2(3.89×104 Da)主要是由中性糖和蛋白质组成的糖蛋白,其糖肽键类型均为O-型糖肽键。HDJS2主要由甘露糖、葡萄糖、半乳糖、阿拉伯糖、岩藻糖和96.1%氨基酸组成;HDJS5-2主要由甘露糖、葡萄糖、半乳糖、阿拉伯糖和71.3%氨基酸组成。抗氧化分析结果表明,HDJS2和HDJS5-2的还原能力、DPPH清除能力和抗氧化能力呈剂量依赖性关系。此外,HDJS2和HDJS5-2均能够促进脾淋巴细胞增殖。结论:HDJS2和HDJS5-2具有潜在的抗氧化作用和免疫调节作用。In this paper,crude glycoprotein from black soybeans(designated HDJ)prepared by alkaline extraction and then alcohol precipitation was fractionated by column chromatography on DEAE-52 cellulose and Sephadex G-100 into two fractions,designated HDJS2 and HDJS5-2.Their chemical compositions were determined and their structures were characterized by spectroscopic methods.Their antioxidant activities were analyzed by reducing power,2,2-diphenyl-1-picrylhydrazyl(DPPH)scavenging rate and total antioxidant capacity assays.Immunoregulatory activities were determined by the methyl thiazolyl tetrazolium(MTT)colorimetric method.The results showed that HDJS2(3.20×104 Da)and HDJS5-2(3.89×104 Da)were mainly composed of neutral carbohydrate and protein.The glycopeptide bond types were O-type.HDJS2 was mainly composed of mannose,glucose,galactose,arabinose and fucose together with 96.1%amino acids.HDJS5-2 was mainly composed of mannose,glucose,galactose and arabinose as well as 71.3%amino acids.HDJS2 and HDJS5-2 had antioxidant activity in a dose-dependent.In addition,both glycoproteins could promote splenocyte proliferation,suggesting that they had immunoregulatory effects.

关 键 词：黑豆 糖蛋白 抗氧化活性 脾细胞增殖 

分 类 号：TS201.1[轻工技术与工程—食品科学]