| 孟乐,金丹青,龚宣伊,等.厚壳贻贝抗氧化肽的分离、鉴定及活性评价[J].中国海洋药物,2024,43(3):59-66. |
| 厚壳贻贝抗氧化肽的分离、鉴定及活性评价 |
| Isolation, identification and activity evaluation of Mytilus coruscus antioxidant peptides |
| 投稿时间：2023-03-10 修订日期：2023-04-26 |
| DOI：10.13400/j.cnki.cjmd.2024.03.005 |
| 中文关键词: 厚壳贻贝 酶解肽 抗氧化活性 分离纯化 |
| English Keywords:Mytilus coruscus hydrolyzed peptides antioxidant activity isolation and purification |
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| 中文摘要: |
| 目的：以厚壳贻贝(Mytilus coruscus)为原材料,以抗氧化活性为筛选指标,运用酶解技术和现代分离纯化技术,制备抗氧化多肽。方法：以1,1-二苯基-2-三硝基苯肼自由基(DPPH?)、羟自由基(HO?)及2,2"-联氮-双-3-乙基苯并噻唑啉-6-磺酸自由基(ABTS?)清除率为指标,优化酶解条件。通过超滤、凝胶过滤层析和反向高效液相色谱分离抗氧化肽,并评价其抗氧化活性。结果：最佳酶解条件为：木瓜蛋白酶、酶解时间3 h、加酶量3%、料液比1：50。通过反相高效液相色谱及测序,鉴定4条多肽分别为HK1：Gln-Thr-Asp(362.34 Da)；HK2：His-Gln-Glu-Glu(541.52 Da)；HK3：Leu-Glu-Gly-Asp-Thr(533.54 Da)；HK4：Thr-Gln-Glu(376.37 Da)。其对DPPH?自由基清除率的EC50分别为0.523±0.002 mg/mL,0.419±0.002 mg/mL,0.467±0.007 mg/mL,0.626±0.011 mg/ mL；对HO?自由基清除率的EC50分别为0.587±0.027 mg/mL,0.866±0.034 mg/mL,1.009±0.018 mg/mL,1.014±0.036 mg/mL)、对ABTS?自由基清除率的EC50分别为0.218±0.001 mg/mL,0.117±0.002 mg/mL,0.121±0.001 mg/mL,0.415±0.003 mg/mL。结论：木瓜蛋白酶酶解厚壳贻贝蛋白,可以产生具有显著自由基清除活性的酶解肽,可为厚壳贻贝抗氧化肽的开发利用提供理论依据和参考。 |
| English Summary: |
| Objective: using antioxidant activities as screening index, to prepare antioxidant peptides from Mytilus coruscus by enzymatic hydrolysis and modern separation and purification technology. Methods: the scavenging rates of DPPH radical (DPPH?) , hydroxyl radical (HO?) and 2,2"-biazo-bis-3-ethylbenzothiazoline-6-sulfonic acid free radical (ABTS?) were measured, to optimize the conditions of enzymatic hydrolysis. Antioxidant peptides were isolated by ultrafiltration, gel filtration chromatography and reverse High-performance liquid chromatography, and their antioxidant activities were evaluated. Results: the optimal conditions for enzymatic hydrolysis were as follows: papain, enzymatic hydrolysis time of 3 h, enzyme dosage of 3% , feed-liquid ratio of 1:50. Four peptides were identified as HK1: Gln-Thr-Asp (362.34 Da); HK2: His-Gln-Glu-Glu (541.52 Da); HK3: Leu-Glu-Gly-Asp-Thr (533.54 Da); HK4: Thr-Gln-Glu (376.37 Da) by reverse high-performance liquid chromatography and amino acid N-terminal sequencing. Their EC50 for DPPH? scavenging were 0.523±0.066 mg/mL, 0.419±0.042 mg/mL, 0.467±0.078 mg/mL, 0.682±0.134 mg/mL, respectively; and their EC50 for HO? scavenging were 0.587±0.150 mg/mL, 0.866±0.123 mg/mL, 1.009±0.060 mg/mL, 1.014±0.122 mg/mL, respectively; the EC50 of O2-? scavenging were 0.239±0.021 mg/mL, 0.245±0.019 mg/mL, 0.611±0.012 mg/mL, 0.716±0.024 mg/mL, respectively; the EC50 of ABTS. scavenging were 0.218±0.109 mg/mL, 0.117±0.013 mg/mL, 0.121±0.019 mg/mL, 0.415±0.054 mg/mL, respectively. Conclusion: enzymatic hydrolysis of Mytilus coruscus protein by papain can produce peptides with significant free radical scavenging activities, which can provide theoretical basis and reference for the development and utilization of Mytilus coruscus. |
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