锦灯笼宿萼多糖的提取纯化及对糖代谢酶的抑制活性

刘玉双; 刘伟; 高佳; 洪瑞; 袁琪; 滕少菲; 孙晨晨; 滕杨

doi:10.13386/j.issn1002-0306.2024120406

锦灯笼宿萼多糖的提取纯化及对糖代谢酶的抑制活性

Extraction and Purification of Polysaccharides from Calyx of Physalis alkekengi L. var. franchetii (Mast.) Makino and Its Effect on the Inhibition of Glucose Metabolic Enzymes

摘要

摘要: 本研究提取纯化锦灯笼宿萼中的多糖（polysaccharides from calyx of Physalis alkekengi L. var. franchetii (Mast.) Makino，PACP）成分，探究其理化性质及降糖活性。在单因素实验的基础上，通过响应面试验优化PACP的最佳提取工艺，经DEAE-52和Sephadex G-100柱层析进行纯化。采用体外抗氧化实验测定纯化多糖对DPPH自由基和ABTS⁺自由基的清除能力，并将清除能力最好的多糖进行定性和定量分析，利用紫外光谱（Ultraviolet Spectroscopy，UV）、高效凝胶渗透色谱（High Performance Gel Permeation Chromatography，HPGPC）、高效液相色谱（High Performance Liquid Chromatography，HPLC）、傅里叶红外光谱（Fourier Transform Infrared Spectroscopy，FT-IR）、三螺旋结构分析和扫描电镜（Scanning Electron Microscope，SEM）对多糖结构进行初步表征，并考察其体外降血糖活性。结果表明，PACP的最佳提取工艺为提取温度90 ℃，液料比36:1 mL/g，提取时间2.2 h，在此条件下的得率为4.47%±0.13%。体外抗氧化实验表明，纯化多糖PACP-1a对DPPH和ABTS⁺自由基清除能力较强，其半数抑制浓度（IC₅₀）分别为0.26和0.31 mg/mL。PACP-1a是一种分子量为109443 Da含有三螺旋结构的β型吡喃糖，其糖含量为88.82%，蛋白质含量为1.29%，糖醛酸含量为28.77%。单糖组成及摩尔比为甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖和木糖组成=0.112:0.025:0.054:0.122:0.054:0.102。PACP-1a对α-淀粉酶和α-葡萄糖苷酶IC₅₀值分别为3.42和3.11 mg/mL，表现出良好的降血糖活性，为锦灯笼宿萼多糖的工业制备及其在功能性食品领域的开发与应用提供理论依据。

Abstract: In this study, exploring the physicochemical properties and hypoglycemic activities of polysaccharides from calyx of Physalis alkekengi L. var. franchetii (Mast.) Makino. Based on the single factor experiment, the best extraction process of PACP was optimized by response surface. The polysaccharide was purified by DEAE-52 and Sephadex G-100 column chromatography. The free radical scavenging ability of the purified polysaccharides was determined in vitro. The polysaccharides with the best scavenging ability were qualitatively and quantitatively analyzed. The structure of the polysaccharide was preliminarily characterized by ultraviolet spectroscopy (UV), high-performance gel permeation chromatography (HPGPC), high performance liquid chromatography (HPLC), fourier transform infrared spectroscopy (FT-IR), triple helix structure analysis and scanning electron microscopy (SEM), and to evaluate its hypoglycemic activity in vitro. Results showed that the optimum extraction conditions of PACP were extraction temperature 90 ℃, liquid-solid ratio 36:1 mL/g and extraction time 2.2 h. Under these conditions, the yield was 4.47%±0.13%. In vitro antioxidant experiments revealed that PACP-1a had a strong effect on DPPH and ABTS⁺, with the half maximal inhibitory concentration (IC₅₀) were 0.26 mg/mL and 0.31 mg/mL, respectively. PACP-1a was a β-pyranose with a molecular weight of 109443 Da and a triple helix structure. The contents of sugar, protein and uronic acid in PACP-1a were 88.82%, 1.29% and 28.77%, respectively. PACP-1a consisted of mannose, rhamnose, galacturonic acid, glucose, galactose and xylose with the molar ratio of 0.112:0.025:0.054:0.122:0.054:0.102. PACP-1a had a good hypoglycemic activity, with the IC₅₀ values of PACP-1a on α-amylase and α-glucosidase were 3.42 mg/mL and 3.11 mg/mL, respectively. This study provides a theoretical basis for the industrial preparation of polysaccharides from calyx of Physalis alkekengi L. var. franchetii (Mast.) Makino and its development and application in the field of functional food.

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