摘要
目的从海茸(Durvillaea antarctica)中提取、分离和纯化β-葡聚糖,并对其结构和体外免疫活性进行研究。方法将干燥海茸粉碎,经水提取和Q-Sepharose Fast Flow离子交换层析分离得到β-葡聚糖(DAC)。采用硫酸-苯酚法、Folin-酚法、柱前衍生高效液相色谱法和多角度高效凝胶渗透色谱与多角度激光光散射器联用(HPGPC-MALLS)法对DAC的总糖含量、蛋白含量、单糖组成和分子量进行分析,并通过甲基化和气质联用(GC-MS)、傅里叶变换红外光谱(FTIR)和核磁共振波谱(NMR)对其结构进行表征。采用中性红法对DAC的体外免疫活性进行评价。结果从海茸中分离纯化得到1种总糖含量达97.5%的β-葡聚糖,其蛋白含量和分子量分别为2.0%和18.3kDa。经甲基化和NMR分析结果表明,DAC是以→3)Glc(β1→为主链,并含有少量→6)Glc(α/β1→分支的葡聚糖。体外免疫活性结果显示,在浓度为50μg·mL-1时,DAC对巨噬细胞的吞噬能力达到最大值。结论海茸β-葡聚糖能显著增强巨噬细胞的吞噬能力,是1种潜在的免疫增强剂。
Objective To extract and purify t3-glucan from Durvillaea Antarctica, and investigate its struc-ture and immune activity. Methods A water soluble glucan was obtained from Durvillaea Antarctica by water extraction at room temperature, and further purified by CaC12 and strong anion exchange chroma-tography. The contents of total sugar and protein, the monosaccharide composition and the molecular mass of the glucan were analyzed by phenol-sulfuric method, folin-phenol method, precolumnderivatiza- tion reversed phase-high performance liquid chromatography （RP-HPLC） and high performance gelper- meation chromatography-multi-angle laser light scattering （HPGPC-MALLS）, respectively. The struc- ture of the g[ucan was characterized by fourier transform infra-red （FTIR） and nuclear magnetic reso- nance （NMR） spectroscopy, methylation analysis and gas chromatography-mass spectrometry （GC- MS） analysis. The immune activity was measured by neutral red method. Results The glucan from Durvillaea Antarctica （DAC） was composed of a backbone of→3）Glc（β1→ and a little branch of -→6）Glc（α/β1→. The immune activity assay showed that 50μg·mL-1 DAC could obviously increase the phagocytosis ability of mononuclear phagocytes in Raw 264. 7 cell line. Conclusion The β-glucan from Durvillaea Antarctica had a significant immune activity in Raw 264. 7 cell line, and could be used as a potential immunity intensifier.
出处
《中国海洋药物》
CAS
CSCD
2015年第5期1-6,共6页
Chinese Journal of Marine Drugs
基金
国家海洋局公益性行业专项(201005024)
国家科技支撑计划项目(2013BAB10B02)资助
关键词
海茸
Β-葡聚糖
结构表征
免疫活性
.DurvillaeaAntarctica
glucan
structure characterizationg immune activity
