微波辅助酶解辣木粕清蛋白制备抗氧化肽及工艺优化

    Preparation of Antioxidant Peptides by Microwave-assisted Enzymatic Hydrolysis of Moringa Meal Albumin and Process Optimization

    • 摘要:
      目的 筛选酶解辣木粕清蛋白的最佳蛋白酶,采用微波辅助酶解制备辣木粕抗氧化肽。
      方法 以辣木粕清蛋白水解度作为考察指标,采用碱性蛋白酶、胰蛋白酶、风味蛋白酶、复合蛋白酶、菠萝蛋白酶以及木瓜蛋白酶进行初筛和复筛,筛选最佳的蛋白酶。通过单因素试验以及响应面试验,筛选最佳酶解条件,采用微波辅助进一步优化酶解工艺。通过测定DPPH·清除率、O2-·清除率、ABTS+·清除率和还原能力评价辣木粕清蛋白抗氧化活性。
      结果 辣木粕清蛋白为酶解的最佳蛋白质,风味蛋白酶为最佳蛋白酶。通过单因素试验以及响应面试验得到最优酶解工艺:加酶量7 491.44 U/g、底物浓度9.36%、pH 7.50、酶解温度45 ℃,水解时间3.88 h,在此酶解条件下,抗氧化肽得率为19.09%,水解度为69.80%。采用低功率微波仪器(0~100 W)进一步将酶解时间缩短到1 h,同时可提高酶解物的抗氧化活性。检测出辣木粕酶解物具有较好的抗氧化活性,清除DPPH·的最佳质量浓度为10 mg/mL;清除ABTS+·、O2-·的最佳质量浓度为50 mg/mL;还原Fe3+的最佳质量浓度为50 mg/mL,与阳性对照Vc相当。
      结论 低功率微波能够提高辣木粕清蛋白酶解效率和抗氧化活性,辣木粕具有制备天然抗氧化肽的潜力。

       

      Abstract:
      Objective The optimal protease for enzymatic hydrolysis of Moringa meal albumin was screened, and the antioxidant peptides of Moringa meal were prepared by microwave-assisted enzymatic hydrolysis.
      Method With the degree of hydrolysis as an index, alkaline protease, trypsin, flavour protease, complex protease, bromelain and papain was used to carry out primary screening and re-screening, and the best protease was screened out. The optimal enzymatic hydrolysis conditions were screened out by single factor experiments and response surface methodology, and the enzymatic hydrolysis process was further optimized by microwave. The antioxidant activity of Moringa meal albumin was evaluated by measuring DPPH· clearance rate, O2-· clearance rate, ABTS+· clearance rate and reducing capacity.
      Result Moringa meal albumin was the best protein for enzymatic hydrolysis, and flavor protease was the optimal protease. By single factor experiments and response surface methodology, the optimal enzymatic hydrolysis process was obtained as: enzyme volume 7 491.44 U/g, substrate concentration 9.36%, pH 7.50, enzymolysis temperature 45℃, and hydrolysis for 3.88 h. Under this enzymatic hydrolysis condition, the yield of antioxidant peptide was 19.09%, and the degree of hydrolysis was 69.80%. The enzymolysis time was further shortened to 1 h by low-power microwave equipment (0-100W), which could also improve the antioxidant activity of the enzymatic hydrolysates. Furthermore, Moringa oleifera meal hydrolysates had good antioxidant activity, the best mass concentrations for scavenging DPPH·, ABTS+· and O2-· were 10 mg/mL, 50 mg/mL and 50 mg/mL respectively, and the best mass concentration for reducing Fe3+ was 50 mg/mL, which was comparable to the positive control Vc.
      Conclusion Moringa meal has the potential to prepare natural antioxidant peptides, while low-power microwave can improve enzymatic hydrolysis efficiency and antioxidant activity.

       

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