蝉花多糖酶法辅助双水相提取工艺优化及其抗氧化、降血糖和降血脂活性分析

李金婷; 钱心燚; 雍一丹; 吴萌萌; 孙华锴; 王雅楠; 陈安徽; 邵颖; 尼再中

doi:10.13386/j.issn1002-0306.2023070233

蝉花多糖酶法辅助双水相提取工艺优化及其抗氧化、降血糖和降血脂活性分析

Optimization of Enzymatic-assisted Aqueous Two-phase Extraction Conditions of Polysaccharides from Cordyceps cicadae and Analysis of Its Antioxidant, Hypoglycemic and Hypolipidemic Properties in Vitro

摘要

摘要: 蝉花多糖是蝉花主要成分之一，具有多种生物活性。本研究以蝉花子实体为原料，采用纤维素酶和果胶酶辅助双水相体系提取蝉花多糖，以期为蝉花多糖的进一步开发利用和深入研究提供参考。通过单因素实验分析了纤维素酶添加量、果胶酶添加量、（NH₄）₂SO₄质量分数以及聚乙二醇（polyethylene glycol，PEG）质量分数对蝉花多糖得率的影响。采用响应面法优化了蝉花多糖的提取工艺，并对其体外抗氧化、降血糖和降血脂生物活性进行了研究。结果表明，蝉花多糖的最优提取条件为：纤维素酶添加量2.01%，果胶酶添加量2.07%，（NH₄）₂SO₄质量分数17.72%，PEG质量分数20.87%，此条件下蝉花多糖得率达到25.34%。在1.0~5.0 mg/mL浓度范围内，蝉花多糖的Fe²⁺螯合率和ABTS⁺自由基清除率最大值分别为16.33%±1.68%和17.38%±3.07%。总还原力测定表明蝉花多糖吸光度最大值为0.53±0.005。另外，蝉花多糖对α-葡萄糖苷酶和α-淀粉酶活性的最大抑制率分别为30.96%±4.53%和49.40%±1.30%。对胆酸钠、牛磺胆酸钠和胆固醇的最大吸附率分别为47.24%±1.89%，44.88%±0.82%和70.19%±2.54%。综上，蝉花多糖具有显著的抗氧化、降血糖和降血脂活性。本研究将为蝉花多糖相关产品的开发，特别是临床应用提供一定的理论依据。

Abstract: Polysaccharides are one of the main constituents of Cordyceps cicadae (C. cicadae), and they show various biological activities. In the present study, C. cicadae fruiting bodies were used as raw materials to extract polysaccharides using an aqueous two-phase system, along with cellulase and pectinase, aiming to provide references for the further development, utilization, and in-depth research of C. cicadae polysaccharides. Effects of cellulase and pectinase addition amounts and (NH₄)₂SO₄ and polyethylene glycol (PEG) mass fractions on the polysaccharide-extraction rates were analyzed by single-factor experiments. The conditions for C. cicadae polysaccharide extraction were optimized by response-surface methodology, and the antioxidant, hypoglycemic, and hypolipidemic properties of the polysaccharides were evaluated in vitro. The optimal conditions for C. cicadae polysaccharide extraction were as follows: Cellulase addition amount 2.01%, pectinase addition amount 2.07%, (NH₄)₂SO₄ mass fraction 17.72%, and PEG mass fraction 20.87%, the polysaccharide-extraction rate reached 25.34% under these conditions. When the C. cicadae polysaccharide concentration was 1.0~5.0 mg/mL, the maximum Fe²⁺-chelating and ABTS⁺ radical-clearance abilities were 16.33%±1.68% and 17.38%±3.07%, respectively. The determination of total reducing power showed that the maximum absorbance of C. cicadae polysaccharides was 0.53±0.005. The maximum inhibitory rates of C. cicadae polysaccharides for α-amylase and α-glucosidase activities were 30.96%±4.53% and 49.40%±1.30% respectively, the maximum adsorption rates of C. cicadae polysaccharides for sodium cholate, sodium taurocholate, and cholesterol were 47.24%±1.89%, 44.88%±0.82%, and 70.19%±2.54% respectively. The results indicated that C. cicadae polysaccharides demonstrate effective antioxidant, hypoglycemic, and hypolipidemic abilities in vitro. This study will provide a theoretical basis for developing C. cicadae polysaccharide-related products, especially for clinical applications.