以金华火腿为研究对象,借助凝胶层析色谱对火腿粗肽进行初步分离,测定各组分的抗氧化性,将样品经由配备在线钠喷离子源的液相色谱串联质谱分析,采用串联EASY-nanoLC 1200的Orbitrap Q-Exactive Plus质谱仪鉴定金华火腿中所含活性肽的序列,发现了31条丰度高且未被鉴定的短肽。并利用PeptideRanker进行预测分析,得到丰度和活性都比较高的3个肽段,氨基酸序列分别为:DHDGPDHW、FPPDVGD、PFGDTH。用DPPH自由基清除能力、ABTS阳离子自由基清除能力和羟自由基清除能力检测抗氧化能力,证明金华火腿粗肽液具有良好的抗氧化性。红外光谱发现粗肽在1 600~1 700 cm-1有吸收带。通过合成短肽,验证肽段的抗氧化能力,分子对接实验表明超氧化物歧化酶与DHDGPDHW、FPPDVGD、PFGDTH均有结合能力,FPPDVGD的结合能力最强为-6.6 kcal/mol。综上,FPPDVGD具有较好的抗氧化能力,可作为天然抗氧化剂应用于食品、药品或化妆品,应用前景非常广泛,为干腌制品的进一步开发利用提供理论支持。
Taking Jinhua ham as the research object, the crude peptide of ham was preliminary separation was carried out by means of gel chromatography to determine the antioxidant capacity of each component.The samples were analyzed by liquid chromatography tandem mass spectrometry equipped with an online sodium jet ion source.The whole system was the Orbitrap Q-Exactive Plus mass spectrometer with EASY-nanoLC 1200 in series to identify the sequence of active peptides in Jinhua ham, and 31 short peptides with high abundance and unidentified were found.PeptideRanker was used for prediction analysis, and three peptides with high abundance and activity were obtained.The amino acid sequence was DHDGPDHW, FPPDVGD, and PFGDTH.Using DPPH free radical scavenging capacity, ABTS cationic scavenging capacity, and · OH free radical scavenging capacity to test antioxidant capacity, it was proved that the Jinhua ham crude peptide solution had good antioxidant capacity.The infrared spectrum showed that the crude peptide had absorption bands from 1 600 cm-1 to 1 700 cm-1.Through the synthesis of short peptides, the antioxidant capacity of the peptide was verified.Molecular docking experiments showed that the superoxide dismutase enzyme had binding capacity with DHDGPDHW, FPPDVGD, and PFGDTH.The FPPDVGD had the strongest binding capacity of -6.6 kcal/mol.The experiment showed that FPPDVGD had a good antioxidant capacity and could be used as a natural antioxidant in food, medicine, or cosmetics.It has a broad application prospect and provides theoretical support for further developing and utilizing dry-cured products.
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