Abstract Currently, most of the polysaccharide fractions are purified using the methods usually limited to the laboratory scale or easily raising question of residual reagent, which may obstruct the future application of polysaccharide. In our study, we employed sequential ultrafiltration with different membranes combined with HPGPC to purify polysaccharide fractions from the water extract of Lentinus edodes directly. The chemical composition, morphology and structure characteristic were analyzed by chemical methods and instrumental analysis including automatic amino acid analyzer, Elemental Analyzer, SEM, HPGPC, HPAEC-PAD, FT-IR and NMR. Results indicated that three polysaccharide fractions named as LE-UF-1, LE-UF-2 and LE-UF-3 were successfully purified using ultrafiltration membranes with MWCO of 2.5, 5 and 10 kDa in sequence. Mw and polysaccharide content of LE-UF-1 (~136 kDa, 86.11%) were the highest, followed by LE-UF-2 (14–61 kDa, 49.82%) and LE-UF-3 (14–35 kDa, 22.44%). Surprisingly, LE-UF-2 had 11.73% amino acid, and LE-UF-3 had 19.64% amino acid. The polysaccharide in all the three fractions was heteropolysaccharide that mainly consisted of Glc, Gal and Man. There might be more β-sugar residues in LE-UF-1 and LE-UF-2, while more α-sugar residues in LE-UF-3. Our study provides a greener and more efficient potential procedure for large-scale polysaccharide purification, and is able to prepare the polysaccharide fractions from Lentinus edodes that may be used for food or drug development.

中文翻译：

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香菇水提物中多糖的膜分离纯化及其化学成分和结构表征

摘要 目前，大部分多糖馏分的纯化方法通常仅限于实验室规模或容易产生残留试剂的问题，这可能会阻碍多糖的未来应用。在我们的研究中，我们采用连续超滤与不同的膜结合 HPGPC 直接从香菇的水提取物中纯化多糖组分。采用氨基酸自动分析仪、元素分析仪、扫描电镜、HPGPC、HPAEC-PAD、FT-IR和NMR等化学方法和仪器分析方法对其化学成分、形貌和结构特征进行分析。结果表明，分别使用 MWCO 为 2.5、5 和 10 kDa 的超滤膜成功纯化了 LE-UF-1、LE-UF-2 和 LE-UF-3 三种多糖组分。LE-UF-1 (~136 kDa, 86.11%) 的 Mw 和多糖含量最高，其次是 LE-UF-2 (14-61 kDa, 49.82%) 和 LE-UF-3 (14-35 kDa, 22.44%)。令人惊讶的是，LE-UF-2 具有 11.73% 的氨基酸，而 LE-UF-3 具有 19.64% 的氨基酸。三个组分中的多糖均为杂多糖，主要由Glc、Gal和Man组成。LE-UF-1 和 LE-UF-2 中的 β-糖残基可能更多，而 LE-UF-3 中的 α-糖残基可能更多。我们的研究为大规模多糖纯化提供了一种更环保、更有效的潜在程序，并且能够从香菇中制备可用于食品或药物开发的多糖组分。LE-UF-3的氨基酸含量为19.64%。三个部分的多糖均为杂多糖，主要由Glc、Gal和Man组成。LE-UF-1 和 LE-UF-2 中的 β-糖残基可能更多，而 LE-UF-3 中的 α-糖残基可能更多。我们的研究为大规模多糖纯化提供了一种更环保、更有效的潜在程序，并且能够从香菇中制备可用于食品或药物开发的多糖组分。LE-UF-3的氨基酸含量为19.64%。三个组分中的多糖均为杂多糖，主要由Glc、Gal和Man组成。LE-UF-1 和 LE-UF-2 中的 β-糖残基可能更多，而 LE-UF-3 中的 α-糖残基可能更多。我们的研究为大规模多糖纯化提供了一种更环保、更有效的潜在程序，并且能够从香菇中制备可用于食品或药物开发的多糖组分。