芝麻抗氧化肽分离纯化与结构鉴定的研究

Separation, Purification and Structure Identification of Antioxidant Peptides Derived from Sesame Protein

通过纳滤、超滤、制备液相对胰蛋白酶水解芝麻蛋白的酶解液中抗氧化肽进行分离纯化,随后采用高效液相质谱联用法进行结构鉴定,并合成相应多肽验证抗氧化活性。结果表明,经分离纯化与结构鉴定,共获得5个芝麻抗氧化肽,Glu-Leu-Phe-Phe-Gly-Ala-Gly-Gly-Glu-Asn-Pro-Glu-Ser-Phe-Phe-Lys(ELFFGAGGENPESFFK)、Phe-Glu-Ser-Glu-Ala-Gly-Leu-Thr-Glu-Phe-Trp-Asp-Arg(FESEAGLTEFWDR)、Asp-Val-Ala-Asn-Glu-Ala-Asn-Gln-Leu-Asp-Leu-Lys(DVANEANQLDLK)、Glu-Asn-IIe-Glu-His-Thr-Ala-Ala-Thr-His-Ser-Tyr-Asn-Pro-Arg(ENIEHTAATHSYNPR)、Gln-Asp-Asn-Ala-Asn-Asn-Ala-Asn-Gln-Leu-Asp-Pro-Asn-Pro-Arg(QDNANNANQLDPNPR)。胰蛋白酶酶解产生的芝麻抗氧化肽N端以酸性氨基酸残基为主,C末端为碱性氨基酸残基;除抗氧化肽ELFFGAGGENPESFFK是芝麻7S酶解产物以外,其余多肽均来自芝麻11S蛋白酶解。

Antioxidant peptides in hydrolysate of sesame protein hydrolyzed by trypsin were isolated by nanofiltration,ultrafiltration and preparative high-performance liquid chromatography successively,then the structure of antioxidant peptide was identified by liquid chromatography-mass spectrometry,finally the antioxidant activity of peptides synthesized based on the result of structure identification was tested.Results showed that 5 antioxidant peptides of sesame were found by means of separation,purification and structure identification,and their sequence were as follows,Glu-Leu-Phe-Phe-Gly-Ala-Gly-Gly-Glu-Asn-Pro-Glu-Ser-Phe-Phe-Lys(ELFFGAGGE-NPESFFK),Phe-Glu-Ser-Glu-Ala-Gly-Leu-Thr-Glu-Phe-Trp-Asp-Arg(FESEAGLTEFWDR),Asp-Val-Ala-Asn-Glu-Ala-Asn-Gln-Leu-Asp-Leu-Lys(DVANEANQLDLK),Glu-Asn-IIe-Glu-His-Thr-Ala-Ala-Thr-His-Ser-Tyr-Asn-Pro-Arg(ENIEHTAATHSYNPR),Gln-Asp-Asn-Ala-Asn-Asn-Ala-Asn-Gln-Leu-Asp-Pro-Asn-Pro-Arg(QDNANNANQLDPNPR).N-terminal amino acids of antioxidant peptides derived from sesame protein hydrolyzed by trypsin were mainly acidic amino acid residuals,while the C terminal amino acids were basic amino acids.Except the antioxidant peptide ELFFGAGGENPESFFK was generated from sesame 7 S globulin,the rest originated from sesame 11 S globulin.