益生菌发酵制备金针菇抗氧化肽的研究

Study on preparation of antioxidant peptides of Flammulina velutipes by probiotics fermentation

试验旨在获得高活性的金针菇抗氧化肽,优化益生菌发酵制备金针菇抗氧化肽的条件。试验以金针菇为发酵基质,以抗氧肽对2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)自由基清除率为指标,对枯草芽孢杆菌、酿酒酵母和黑曲霉3种益生菌进行了筛选,使用pH值沉淀法分离抗氧化肽的最适pH值,采用单因素和响应面试验对发酵条件(液料比、发酵时间、发酵温度、金针菇用量)进行考察。结果显示,枯草芽孢杆菌适合发酵金针菇制备抗氧化肽;沉淀抗氧肽的最适pH值为6;最佳发酵条件为液料比4.50 mL/g、发酵时间30 h、发酵温度35℃、金针菇用量40.00 g,此条件下金针菇抗氧化肽对ABTS自由基清除率预测值为95.60%,实测值为96.75%,实测值与预测值相差很小,表明此响应面法优化的工艺可靠,截留分子量小于3 kD的抗氧肽活性最强。研究表明,通过此法制备的金针菇抗氧肽具有很强的抗氧化性,为金针菇产品的开发利用提供了参考。

The study was to obtain highly active Flammulina velutipes antioxidant peptides,optimized the conditions for preparation of Flammulina velutipes antioxidant peptides by fermentation of probiotics.In the test,Flammulina velutipes was used as the fermentation substrate,and the free radical scavenging rate of ABTS was used as an indicator.Three probiotics,Bacillus subtilis,Saccharomyces cerevisiae and Aspergillus niger were screened,and the optimum pH value for precipitation method for isolating antioxidant peptides was optimized.The fermentation conditions(liquid to material ratio,fermentation time,fermentation temperature,amount of Flammulina velutipes)were investigated by response surface methodology.The results showed that Bacillus subtilis was the most suitable for fermenting Flammulina velutipes to prepare antioxidant peptides,the optimum pH value for precipitating antioxidant peptides was 6,the optimal fermentation conditions were liquid to material ratio of 4.50 mL/g,and the fermentation time of 30 h,the fermentation temperature of 35℃,and the amount of Flammulina velutipes of 40.00 g.Under these conditions,the predicted value of Flammulina velutipes antioxidant peptide to free radical scavenging rate of ABTS was 95.60%,and the measured value was 96.75%.The deviation between the measured value and the predicted value was very small,indicating that the optimized process by the response surface method was reliable.The antioxidant peptide with molecular weight cut-off less than 3 kD had the strongest activity.The study indicates that the antioxidant peptide of Flammulina velutipes prepared by the method had strong antioxidant activity,which provided a reference for the development and utilization of flammulina velutipes products.