Study on Isolation, Purification, Structure Characterization and Antioxidant Activity of Polysaccharide from Zizyphus jujuba cv.
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摘要: 本实验以骏枣为原料，对骏枣多糖的分离纯化、结构表征及抗氧化活性进行研究。通过水提醇沉得骏枣粗多糖HZPC，再经DEAE-52阴离子层析柱和Sephadex G-100葡聚糖凝胶柱分离纯化获得精制骏枣多糖组分HZPC-2。利用高效凝胶色谱法（HPGPC）和离子色谱法（IC）测定HZPC-2的分子量及单糖构成，紫外光谱、红外光谱和扫描电镜研究HZPC-2的结构特征，最后对HZPC-2的抗氧化活性进行评估。结果表明：HZPC-2为α-吡喃葡萄苷骨架的中性多糖（JDP-N），分子量为3.25×104 Da，是由鼠李糖（Rha）、阿拉伯糖（Ara）、半乳糖（Gal）、葡萄糖（Glc）、木糖（Xyl）、甘露糖（Man）和半乳糖醛酸（GalA）构成，其摩尔比为0.05:0.34:0.29:0.15:0.08:0.02:0.06。扫描电子显微镜观察结果显示，HZPC-2表面呈沟壑状且朝四面延伸，四周嵌着孔状结构。体外抗氧化试验表明，HZPC-2对三种自由基清除活性的最强的是DPPH自由基，其次是羟自由基，最弱的是ABTS自由基，这可作为潜在抗氧化剂以及为开发具有骏枣多糖功能的食品提供研究基础。Abstract: The isolation and purification, structure characterization and antioxidant activity of Zizyphus jujuba cv. Junzao polysaccharides were studied. The crude polysaccharide HZPC was extracted from Zizyphus jujuba cv. Junzao by water extraction and ethanol precipitation, and then separated and purified by DEAE-52 anionic chromatography column and Sephadex G-100 dexan gel column. The molecular weight and monosaccharide composition of HZPC-2 were determined by high performance gel chromatography (HPGPC) and ion chromatography (IC), and the structural characteristics of HZPC-2 were studied by ULTRAVIOLET spectrum, infrared spectrum and scanning electron microscope. Finally, the antioxidant activities of HZPC-2 were evaluated. The results showed that HZPC-2 polysaccharide was a neutral polysaccharide with α-glucopyranoside framework (JDP-N), its molecular weight was 3.25×104 Da. It was composed of rhamnose (Rha), arabinose (Ara), galactose (Gal), glucose (Glc), xylose (Xyl), mannose (Man) and galacturonic acid (GalA). The mole ratio was 0.05:0.34:0.29:0.15:0.08:0.02:0.06. Scanning electron microscopy showed that the surface of HZPC-2 was furry-shaped and extended in all directions, and the interior of HZPC-2 was embedded with porous structures. In vitro antioxidant tests showed that HZPC-2 had the strongest scavenging activity against DPPH radical, followed by hydroxyl radical, and the weakest scavenging activity against ABTS radical. It can be used as a potential antioxidant and provide a basis for the development of food with the function of Zizyphus jujuba cv. Junzao polysaccharides.
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图 3 HZPC-2的紫外吸收光谱（a）和傅里叶红外吸收光谱图（b）
Figure 3. UV (a) and FTIR (b) absorption spectra of HZPC-2
图 4 HZPC-2单糖构成分析
Figure 4. Analysis of monosaccharide composition of HZPC-2
注：Rha：鼠李糖；Ara：阿拉伯糖；Gal：半乳糖；Glc：葡萄糖；Xyl：木糖；Man：甘露糖；GalA：半乳糖醛酸。
表 1 HZPC-2单糖构成分析
Table 1. Analysis of monosaccharide composition of HZPC-2
组别 标准品 HZPC-2样品 摩尔比 保留时间
（min）峰面积
（%）保留时间
（min）峰面积
（%）岩藻糖 Fuc 6.05 7.97 6.05 0.00 − 盐酸氨基半乳糖 GalN 11.27 10.76 11.27 0.00 − 鼠李糖 Rha 11.76 8.18 11.76 1.19 0.05 阿拉伯糖 Ara 12.63 6.55 12.63 7.49 0.34 盐酸氨基葡萄糖 GlcN 14.09 21.16 14.09 0.00 − 半乳糖 Gal 15.93 8.20 15.93 7.23 0.29 葡萄糖 Glc 18.03 10.02 18.03 4.48 0.15 N-乙酰-D氨基葡萄糖 GlcNAc 19.66 3.21 19.66 0.00 − 木糖 Xyl 21.03 5.79 21.03 1.17 0.08 甘露糖 Man 21.62 6.27 21.62 0.44 0.02 果糖 Fru 24.86 5.46 24.86 0.00 − 核糖 Rib 27.76 18.86 27.76 0.00 − 半乳糖醛酸 GalA 45.42 8.42 45.42 1.65 0.06 古罗糖醛酸 GulA 46.15 13.25 46.15 0.00 − 葡萄糖醛酸 GlcA 48.63 10.82 48.63 0.00 − 甘露糖醛酸 ManA 50.98 25.71 50.98 0.00 − 
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