鲍鱼脏器碱提多糖的分离纯化、结构表征及抗氧化活性研究

林志超; 潘晓明; 吴启赐; 薛钰; 黄家福; 潘裕添

doi:10.13386/j.issn1002-0306.2023030266

鲍鱼脏器碱提多糖的分离纯化、结构表征及抗氧化活性研究

Isolation, Purification, Structure Characterization and Antioxidant Activity of Alkali-extracted Polysaccharide from Abalone Viscera

摘要

摘要: 目的：分离纯化鲍鱼脏器碱提多糖（alkali-extracted abalone vscera polysaccharides，Aavp），并研究其结构和抗氧化活性，以期为鲍鱼脏器碱提多糖的开发及应用提供参考。方法：通过热碱提醇沉获得鲍鱼脏器碱提粗多糖，经DEAE Sepharose Fast Flow和Sephacryl S-400 HR纯化得到高纯度多糖，并利用气相色谱、高效液相色谱、红外光谱和热重分析仪等工具分析其结构及抗氧化活性。结果：粗多糖纯化后得到四个组分，包括Aavp Ia、Aavp Ib、Aavp IIa和Aavp IIb，其中Aavp IIa得率较高，选择其进一步进行结构分析。分析发现，Aavp IIa由木糖和半乳糖组成，分子量为166513 Da；糖苷键构型为α型，组成如下：半乳糖1→4糖苷键摩尔百分比为11.81%，半乳糖1→3糖苷键为34.14%，半乳糖1→2糖苷键为10.14%；木糖1→3糖苷键摩尔百分比为33.85%，1→2和1→4糖苷键共占10.06%。热重分析显示，Aavp IIa在226.4-332.6 ℃下断裂分解，热失重率为43.65%。抗氧化实验显示Aavp IIa对O₂⁻·的清除率为85.89%，对DPPH·的清除率为62.17%，具有一定的抗氧化活性。结论：提取的鲍鱼脏器碱提多糖是一类杂多糖，并具有一定的抗氧化活性。

Abstract: Objective: Alkali-extracted polysaccharides from Abalone viscera (Aavp) were isolated and purified. The structure and antioxidant activity of Aavp were studied, which would provide a reference for developing and applying polysaccharides. Methods: The crude alkali-extracted polysaccharide was prepared by hot alkali extraction and alcohol precipitation. After the purification of DEAE Sepharose Fast Flow and Sephacryl S-400 HR, the purified polysaccharide was obtained and subjected to the structural analysis and antioxidant activity assay, such as Gas chromatography (GC), high-performance liquid chromatography (HPLC), infrared spectroscopy (IR), and thermogravimetric analyzer (TGA), etc. Result: Four kinds of components (Aavp Ia, Aavp Ib, Aavp IIa, and Aavp IIb) were obtained from the crude polysaccharide. Because of the highest yield, Aavp IIa was selected for further structural analysis. Aavp IIa was composed of xylose and galactose, with a relative molecular weight of 166513 Da. IR characterized the α-glycosidic bond configuration. The composition of Aavp IIa was possible as follows: The molar percentage of the galactose 1→4 glycosidic bond was 11.81%, the galactose 1→3 glycosidic bond was 34.14%, and the galactose 1→2 glycosidic bond was 10.14%. The molar percentage of the xylose 1→3 glycosidic bond was 33.85%, and the 1→2 and 1→4 glycosidic bond was 10.06%. Thermogravimetric analysis showed that Aavp IIa was broken and decomposed at 226.4~332.6 °C, and the thermal weight loss rate was 43.65%. Antioxidant experiments showed that Aavp IIa had a scavenging rate of 85.89% for superoxide anion radicals and 62.17% for DPPH radicals, respectively, presenting a certain antioxidant activity. Conclusion: Aavp is a heteropolysaccharide with specific antioxidant activity.