羊肚菌多糖的提取优化及抗氧化活性研究

Study on the extraction optimization and antioxidant activity of Morchella esculenta polysaccharides

中文关键词:  羊肚菌多糖  热水浸提法  柠檬酸提取法  碱提法  低共熔溶剂提取法  响应面法  乙醇分级  抗氧化活性

英文关键词:Morchella esculenta polysaccharides  hot water extraction method  citric acid extraction method  alkaline solution extraction method  deep eutectic solvents extraction method  response surface methodology  ethanol fractionation  antioxidant activity

中文摘要:

      目的 对羊肚菌多糖(Morchella esculenta polysaccharides, MEP)的提取工艺进行筛选优化, 获得最佳工艺和最佳活性组分。方法 采用热水浸提(hot water extraction, HWE)法、柠檬酸提取(citric acid extraction, CAE)法、碱提(alkaline solution extraction, ASE)法、低共熔溶剂(deep eutectic solvents, DESs)提取法获得4种多糖(MEP-W、MEP-S、MEP-A和MEP-D), 筛选最佳提取方法, 在单因素试验基础上通过响应面设计优化最佳工艺, 采用分级醇沉法分离纯化MEP-W获得3个组分MEP30、MEP60和MEP80, 通过自由基清除试验和还原力测定评估其抗氧化活性。结果 HWE法为最佳提取方法, 最佳提取工艺为: 提取时间3.0 h、液料比44:1 (mL/g)、温度83.6℃, 此时的提取得率为(8.13±0.38)%。MEP80在3个组分中清除·OH能力和还原力最强, 半数效应浓度(median effective concentration, EC50)和RP0.5AU分别为0.44和1.52 mg/mL, 高于其前体物MEP-W。结论 热水浸提为最佳方法, MEP80为抗氧化活性的最佳组分, 在开发功能性食品和医药行业应用中值得进一步研究。

英文摘要:

      Objective To screen and optimize the extraction process of Morchella esculenta polysaccharides (MEP), and obtain the best process and the best active components. Methods Four kinds of corresponding polysaccharides (MEP-W, MEP-S, MEP-A and MEP-D) were obtained by hot water extraction (HWE) method, citric acid extraction (CAE) method, alkaline solution extraction (ASE) method, and deep eutectic solvents (DESs) extraction method, the optimal method was selected. On the basis of single factor test, the optimal process was optimized by response surface design. MEP-W fractions MEP30, MEP60 and MEP80 were isolated and purified by fractional alcohol precipitation method, and their antioxidant activity was evaluated by free radical scavenging assay and reducing power assay. Results HWE method was the best method, and the best extraction process was as follows: Extraction time 3.0 h, liquid-to-material ratio 44:1 (mL/g), temperature 83.6℃. Under this condition, extraction yield was (8.13±0.38)%. MEP80 exhibited the strongest antioxidant activity based on hydroxyl radical-scavenging ability and reducing force assay among the 3 components, and its median effective concentration (EC50) and RP0.5AU were 0.44 and 1.52 mg/mL, respectively. Conclusion HWE method is the optimal method, and MEP80 is the strongest in terms of antioxidant activity, which is worth further study in functional food development and pharmaceutical industry.