冠县灵芝多糖的分离纯化、结构表征及抗氧化活性研究

刘养山; 张景; 任洪飞; 逄梦玉; 隋昕怡; 杜秀菊

doi:10.13386/j.issn1002-0306.2022080313

冠县灵芝多糖的分离纯化、结构表征及抗氧化活性研究

doi: 10.13386/j.issn1002-0306.2022080313

聊城大学生命科学学院，山东聊城 252000

基金项目: 山东省自然科学基金项目（ZR2015CL002）；聊城大学科研基金项目（318011707）。

详细信息

作者简介:
刘养山（1987−），男，硕士研究生，研究方向：天然产物的结构与功能，E-mail：liuyangshan0212@163.com

通讯作者:
杜秀菊（1967−），女，博士，教授，研究方向：食药用菌活性成分的结构与功能，E-mail：wyxx2008@163.com

中图分类号: TS201.4
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- 文章访问数: 69
- HTML全文浏览量: 25
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- 被引次数: 0
出版历程
- 收稿日期: 2022-08-31
- 网络出版日期: 2023-05-16
- 刊出日期: 2023-06-15

Extraction, Purification, Structural Characterization and Antioxidant Activity of Polysaccharides from the Fruiting Body of Guanxian Ganoderma lucidum

College of Life Sciences, Liaocheng University, Liaocheng 252000, China

摘要

摘要: 以冠县灵芝为研究对象，采用传统浸提法获得水溶性灵芝多糖GLP，经Sevag法去蛋白及活性炭脱色，然后依次采用Capto^TM DEAE离子柱层析和Superdex 6 prep Grad凝胶柱层析进行分离纯化，获得均一多糖GLPS80a，经高效凝胶色谱（HPGPC）法检测，其相对分子量为9024 Da，高效阴离子色谱（HPAEC）、红外光谱（FT-IR）和核磁共振（NMR）图谱分析结果表明，GLPS80a是由岩藻糖（Fuc）、氨基葡萄糖（GlcN）、半乳糖（Gal）、葡萄糖（Glc）、木糖（Xyl）、甘露糖（Man）和葡萄糖醛酸（GluA）等7种单糖组成，摩尔比为0.06:0.23:0.17:1.00:0.08:0.19:0.23，主要以β-D-（1,6）糖苷键连接的吡喃糖为主要组成且不含三股螺旋的酸性多糖。抗氧化结果表明，GLPS80a具有一定的DPPH自由基、OH自由基和ABTS⁺自由基的清除能力，它们的EC₅₀分别为7.40、8.74和0.33 mg/mL，还原力RP_0.5AU为8.33 mg/mL。本研究将为合理开发冠县灵芝资源和精深加工奠定理论基础。
- 灵芝 /
- 多糖 /
- 分离纯化 /
- 结构表征 /
- 抗氧化活性
Abstract: Taking Ganoderma lucidum in Guanxian County as the experimental material, the crude polysaccharide GLP was extracted from the fruiting body of G. lucidum by hot water. After deproteinization by Sevag methods and decolorization by activated carbon methods, and then separation and purification by Capto^TM DEAE ion chromatography and Superdex 6 prep Grad gel chromatography, respectively, the homogeneous polysaccharide GLPS80a was obtained. And its average molecular weight was 9024 Da. Its chemical structure was preliminarly characterized by monosaccharide composition analysis, infrared scanning and nuclear magnetic resonance analysis, respectively. The results showed that GLPS80a was composed of fucose, glucosamine, galactose, glucose, xylose, mannose and gluconal, and the molar percentage was 0.06:0.23:0.17:1.00:0.08:0.19:0.23. It was an acid polysaccharide with the main of pyranoid glucose linked by β-D-(1,6) glycosidic bond and did not have a triple helix structure. The antioxidant results indicated that GLPS80a had scavenging ability of DPPH radical, ·OH radical and ABTS⁺ radical. Their EC₅₀ was 7.40, 8.74 and 0.33 mg/mL, respectively, and the reducing power RP_0.5AU was 8.33 mg/mL. This work would lay a theoretical foundation for development of G. lucidum resources in Guanxian County.
- Ganoderma lucidum /
- polysaccharides /
- separation and purification /
- structural characterization /
- antioxidant activity

HTML全文

图 1 GLP通过Capto^TM DEAE离子层析柱的洗脱曲线

Figure 1. Elution curve of GLP by CaptoTM DEAE ion chromatography column

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图 2 GLPS80经Superdex 6 prep Grad凝胶柱洗脱曲线

Figure 2. Elution curve of GLPS80 on Superdex 6 prep Grad gel column

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图 3 GLPS80a HPGPC 色谱图（a）和葡聚糖标准品的标准曲线（b）

Figure 3. GLPS80a HPGPC chromatogram (a) and the standard curve for dextran (b)

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图 4 11种标准单糖和GLPS80a单糖组成HPAEC-PAD色谱图

Figure 4. HPAEC-PAD chromatograms of eleven standard monosaccharides and GLPS80a

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图 5 GLPS80a的傅里叶变换红外光谱图

Figure 5. Fourier transform infrared spectra of GLPS80a

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图 6 GLPS80a¹H 核磁共振谱图

Figure 6. ¹H NMR spectra of GLPS80a

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图 7 GLPS80a¹³C核磁共振谱图

Figure 7. ¹³C NMR spectrum of GLPS80a

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图 8 GLPS80a与刚果红混合溶液的最大吸收波长在不同NaOH浓度下的变化情况

Figure 8. Maximum absorption wavelength of the mixed solution of GLPS80a and Congo red at different concentrations of NaOH

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图 9 灵芝多糖清除DPPH自由基能力

Figure 9. DPPH free radical-scavenging ability of Ganoderma lucidum polysaccharides

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图 10 灵芝多糖清除·OH自由基能力

Figure 10. ·OH free radical-scavenging ability of Ganoderma lucidum polysaccharides

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图 11 灵芝多糖的还原力

Figure 11. Reducing power of Ganoderma lucidum polysaccharides

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图 12 灵芝多糖GLPS80a及其前体物GLP对ABTS⁺自由基的清除能力

Figure 12. ABTS⁺ free radical-scavenging activity of GLPS80a and GLP

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表 1 离子交换柱层析样品糖含量和得率

Table 1. Sugar content and yield of ion-exchange column chromatography samples

样品	物理性状	总糖含量（%）	还原糖含量（%）	多糖含量（%）	蛋白含量（%）	得率（%）
GLPW	白色絮状	34.40±0.72	4.40±0.08	30.00±0.60	0.15±0.01	6.29
GLPS20	红棕色片状	57.10±0.86	2.60±0.03	54.50±0.53	0.29±0.01	17.22
GLPS40	淡黑色粉末	25.12±0.31	0.70±0.01	24.42±0.36	0.25±0.01	5.40
GLPS60	淡黑色粉末	21.30±0.26	0.70±0.01	20.60±0.33	0.16±0.01	5.17
GLPS80	浅咖啡色片状	19.84±0.39	*	19.84±0.69	0.15±0.01	4.63
GLPS100	咖啡色片状	11.94±0.20	*	11.94±0.37	0.16±0.01	2.08
注：*表示未测出。

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