Optimization of ultrasonic pretreatment assisted enzymatic hydrolysis for preparation of antioxidant peptides from Flammulina velutipes by response surface methodology
In order to obtain stronger antioxidant peptides from Flammulina velutipes,the paper optimized the optimal process conditions for the preparation of Flammulina velutipes antioxidant peptides using ultrasonic pretreatment assisted enzymatic hydrolysis.In the experiment,the scavenging rate of Flammulina velutipes peptide against the free radical of 2,2-diazobio bis(3-ethyl-benzothiazole-6-sulfonic acid)diamine salt(ABTS)was taken as the index,papain,trypsin,neutral protease and alkaline protease were screened,and the optimal pH for the separation of antioxidant peptides by the isoelectric precipitation method was optimized.Then a single factor experiment was conducted on the basis of determining trypsin as the optimal protease and 6 as the optimal pH for the precipitation of antioxidant peptides.A four factor and three level response surface experiment was conducted under the premise of a fixed liquid to material ratio of 40∶1(mL/g),enzymatic hydrolysis temperature of 37 ℃,and enzymatic hydrolysis pH of 8 to explore the effects of ultrasound power,ultrasound time,enzyme addition amount,and enzymatic hydrolysis time on the preparation process.Finally,the classification and activity comparison of antioxidant peptides were conducted through ultrafiltration separation method.The results showed that the optimal preparation conditions for the antioxidant peptide of Flammulina velutipes were:ultrasound time of 6 minutes,ultrasound power of 296 W,enzyme addition amount of 1.1％(g/g)of the sample amount of Flammulina velutipes,enzymatic hydrolysis time of 75 minutes.Under these conditions,the clearance rate of Flammulina velutipes antioxidant peptide on ABTS was(76.63±1.24)％;Peptides with a molecular weight of＜3 kDa had the highest clearance rate for ABTS·.This indicated that the peptide from Flammulina velutipes had certain antioxidant properties,providing a basis for the development of antioxidant peptides from Flammulina velutipes.
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