摘要
本文通过羊肚菌发酵豆渣生产多糖,采用DEAE-50纤维素柱层析和G-100葡聚糖凝胶层析柱对粗多糖进行分离纯化,得到3种多糖组分,命名为MP-1,MP-3和MP-4,其提取率分别为26.2%,29.1%和18.7%。围绕纯化后的三种多糖进行研究,紫外扫描结果显示,260-280nm之间并没有吸收峰,表明MP-1,MP-3,MP-4中不舍核酸和蛋白质。红外光谱结果表明,三种多糖的结构中均存在β-糖苷键。通过GC-MS对MP-1,MP-3,MP-4的单糖组成进行检测,结果显示,组成的三种多糖的单糖主要有葡萄糖(glucose)、甘露糖(mannose)以及半乳糖(galactose),还含有少量的阿拉伯糖(araribose),鼠李糖(rhamnose)和木糖(xylose)。通过MTT实验可以得知,MP-3对小鼠巨噬细胞有明显的增殖作用,且与浓度呈量效关系,在25μg/mL时为313.57%。浓度为50μg/mL的MP-1对人体肝癌细胞增殖抑制率为68.01%。这一研究发现对多糖的抗肿瘤的机制指明了方向,并为功能性食品因子或药物的开发提供了理论依据。
Crude polysaccharides(MPS) from soybean residue fermented with Morchella esculenta were extracted and purified by DEAE Sephadex A-50 chromatography and Sephadex G-100 size-exclusion chromatography in sequence.Three main fractions MP-1,MP-3 and MP-4 were obtained during the purification steps.The recovery rates based on MPS used were 26.2%,29.1%and 18.7%for MP-1,MP-3 and MP-4 respectively.The monosaccharide composition,ultraviolet spectrum,infrared spectrum and NMR of the three fractions were analyzed in this study.Furthermore,the influence of polysaccharides fractions upon activation of macrophage cells(RAW 264.7) and antitumor activities in the human hepatocarcinoma cell line(HepG-2)in vitro were evaluated.The results indicated that the proliferation of MP-3 on RAW 264.7 was 313.57%at 25μg/mL,which is high while MP-1 had a higher growth inhibition effect on HepG-2 cells of 68.01%at concentration of 50μg/mL.The purified polysaccharides of MP-1 and MP-3 can be used for functions related to the development of food and medicine.
引文