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目的:从人参根中分离纯化人参糖肽组分P-Ⅱ并研究其理化性质。方法:首先采用50%乙醇提取,高浓度乙醇沉淀,活性炭脱色得到人参糖肽,经DEAE-Sepharose CL-6B柱分离,依次用0.2,0.5,1.0 mol·L-1的Na Cl水溶液洗脱,得到组分Ⅱ,然后利用Sephadex G-150柱分离,以0.2 mol·L-1的Na Cl水溶液洗脱得到P-Ⅱ;以葡萄糖、半乳糖醛酸和牛血清白蛋白为对照品测定中性糖、糖醛酸及肽的含量,以醋酸纤维薄膜层析、Sephadex G-100柱色谱、气相色谱分析仪、氨基酸分析仪对P-Ⅱ性质进行研究。结果:P-Ⅱ含中性多糖50.2%,糖醛酸38.5%,肽1.9%,P-Ⅱ为均一糖肽,糖部分的单糖组成为Rha,Ara,Glc和Gal,其摩尔比为0.45∶1.15∶1.00∶2.36,肽部分由Asp,Fhr,Ser等16种氨基酸组成。结论:分离得到的P-Ⅱ为均一糖肽,而不是糖和肽的混合物,为人参糖肽构效学研究奠定基础。
OBJECTIVE: To isolate and purify P-Ⅱ from ginseng root and study its physicochemical properties. Methods: Ginseng glycopeptide was obtained by 50% ethanol extraction, high concentration ethanol precipitation and activated carbon decolorization. The glycopeptide was separated by DEAE-Sepharose CL-6B column and eluted with 0.2, 0.5 and 1.0 mol·L-1 NaCl aqueous solution, The fraction Ⅱ was obtained and then separated by Sephadex G-150 column and eluted with 0.2 mol·L-1 NaCl aqueous solution to obtain P-Ⅱ. Glucose, galacturonic acid and bovine serum albumin , Uronic acid and peptide content, the properties of P-Ⅱ were studied by cellulose acetate membrane chromatography, Sephadex G-100 column chromatography, gas chromatograph and amino acid analyzer. RESULTS: P-Ⅱ contained 50.2% of neutral polysaccharides, 38.5% of uronic acids and 1.9% of peptides, and P-Ⅱ was a homogeneous glycopeptide. The monosaccharide composition of the sugar moiety was Rha, Ara, Glc and Gal with a molar ratio of 0.45 : 1.15: 1.00: 2.36, the peptide part consists of 16 kinds of amino acids such as Asp, Fhr, Ser. Conclusion: The isolated P-Ⅱ is a homogeneous glycopeptide, not a mixture of sugar and peptide, which lays the foundation for the study of the ganoderma lucidum pharmacophore.