刘欢,戚怡,唐晓宁,等.方格星虫酶解水溶性多肽的抗氧化活性及多肽结构鉴定[J].中国海洋药物,2021,(6):-.
方格星虫酶解水溶性多肽的抗氧化活性及多肽结构鉴定
Antioxidant activity and structural identification of enzymatic hydrolysis peptides from Sipunculus nudus L
投稿时间:2021-05-26  修订日期:2021-07-13
DOI:
中文关键词:  方格星虫  酶解  水溶性多肽  抗氧化活性  构-效关系
English Keywords:Sipunculus nudus L  enzymolysis  peptides  antioxidant activity  structure-activity relationship
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作者单位邮编
刘欢 广东医科大学 药学院 524023
戚怡 广东湛江海洋医药研究院 
唐晓宁 广东医科大学 药学院 
林芊杏 广东医科大学 药学院 
吕应年 广东医科大学 药学院 524023
罗辉 广东医科大学 药学院 
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中文摘要:
      目的 以方格星虫为原料,酶解制备水溶性多肽并考察其抗氧化活性,探究其结构-活性关系。方法 利用5种蛋白酶酶解新鲜方格星虫,并结合超滤方法得到高分子量(>50kDa)、中分子量(5~50 kDa)和低分子量(<5kDa)的酶解肽段;利用氨基酸自动分析仪测定方格星虫酶解肽的氨基酸组成。综合考察多肽对1,1-二苯基-2-苦基肼(DPPH.)自由基、羟基自由(.OH)、超氧阴离子自由基( O-2.)的清除能力以及还原力效果,评价方格星虫多肽抗氧化活性;利用高效液相色谱-串联质谱方法鉴定小分子肽的氨基酸组成和序列结构。结果 胰蛋白酶水解方格星虫4h得到的酶解液蛋白含量最高,并在酶解液中检测确定了17种氨基酸,其中精氨酸含量最高13.65%。高中低三组肽段中,低分子量肽段的抗氧化活性优于其他:对DPPH 自由基和羟自由基清除能力大小顺序为低分子量>高分子量>中分子量、对超氧阴离子清除能力大小顺序为低分子量>高分子量≈ 中分子量、还原能力强弱顺序为高分子量>低分子量 > 中分子量。经高效液相色谱-串联质谱联鉴定了GFAGDDAPR、GLGGLSPEK 、LDLAGR和VTKEIPR 4个肽段。结论 低分子量的方格星虫酶解多肽具有较好的抗氧化活性,有潜力用作生物医药或化妆品工业的原料。
English Summary:
      Objective To prepare water-soluble peptides by enzymatic hydrolysis from Sipunculus nudus , and investigate these peptides antioxidant activity-chemical structure relationship. Methods Five kinds of proteases were used to enzymolysis fresh Sipunculus nudus and combined with ultrafiltration method to obtain larger molecular weight (>50kDa), medium molecular weight (5-50 kDa) and smaller molecular weight (<5kDa) peptides; automatic amino acid analyzer was used to determine amino acid composition of the enzymatic peptides; According to 1,1-diphenyl-2-picrylhydrazine (DPPH.) free radicals, hydroxyl free (.OH) , Superoxide anion free radical (O-2.) scavenging ability and reducing power, the results of the anti-oxidative activity of Sipunculus nudus peptides were evaluated ; high performance liquid chromatography- tandem mass spectrometry was used to identify the amino acid sequences of smaller molecule peptides. Results The protein content of the Sipunculus nudus hydrolysate obtained by trypsin hydrolysis of for 4 hours was the highest, and 17 kinds of amino acids were determined the hydrolysate,the highest arginine content is 13.65%. Among the three groups of larger, medium and smaller molecular weight peptides, the antioxidant activity of smaller molecular weight peptides is better than others: the order of scavenging capacity for DPPH and hydroxyl free radical is smaller molecular weight>larger molecular weight>medium molecular weight, and the order of superoxide anion free radical scavenging ability is smaller molecular weight>larger molecular weight medium molecular weight, and the order of reducing ability is larger molecular weight>smaller molecular weight>medium molecular weight, Four peptides were identified as GFAGDDAPR, GLGGLSPEK, LDLAGR and VTKEIPR by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS). Conclusion The smaller molecular weight Sipunculus nudus peptide has good antioxidant activity and has the potential to be used as a raw material for biomedicine or cosmetics industry.
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