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.
[1] 李轻舟, 王红育.发酵肉制品研究现状及展望[J].食品科学, 2011, 32(3):247-251.
LI Q Z, WANG H Y.Present status and prospects of research on fermented meat products[J].Food Science, 2011, 32(3):247-251.
[2] VIRGILI R, SACCANI G, GABBA L, et al.Changes of free amino acids and biogenic amines during extended ageing of Italian dry-cured ham[J].LWT - Food Science and Technology, 2007, 40(5):871-878.
[3] MORA L, GALLEGO M, ARISTOY M C, et al.Peptides naturally generated from ubiquitin-60S ribosomal protein as potential biomarkers of dry-cured ham processing time[J].Food Control, 2015, 48:102-107.
[4] JIMÉNEZ-COLMENERO F, VENTANAS J, TOLDRÁ F.Nutritional composition of dry-cured ham and its role in a healthy diet[J].Meat Science, 2010, 84(4):585-593.
[5] MORA L, CALVO L, ESCUDERO E, et al.Differences in pig genotypes influence the generation of peptides in dry-cured ham processing[J].Food Research International, 2016, 86:74-82.
[6] 张俊艳, 贺阳.超氧化物歧化酶研究与应用[J].食品工业, 2012, 33(3):119-122.
ZHANG J Y, HE Y.Literature review of research and application on superoxide dismutase[J].The Food Industry, 2012, 33(3):119-122.
[7] 郑锦晓, 邢路娟, 周光宏, 等.三种中国传统干腌火腿中粗多肽的抗氧化与抑菌活性的比较[J].食品工业科技, 2018, 39(16):75-79;86.
ZHENG J X, XING L J, ZHOU G H, et al.Antioxidant and antibacterial abilities of crude peptides extracted from the three Chinese traditional dry-cured hams[J].Science and Technology of Food Industry, 2018, 39(16):75-79;86.
[8] BRAND-WILLIAMS W, CUVELIER M E, BERSET C.Use of a free radical method to evaluate antioxidant activity[J].LWT - Food Science and Technology, 1995, 28(1):25-30.
[9] BANGAR S P, SANDHU K S, TRIF M, et al.The effect of mild and strong heat treatments on in vitro antioxidant properties of barley (Hordeum vulgare) cultivars[J].Food Analytical Methods, 2022, 15(8):2193-2201.
[10] 那吉, 马娇, 张海芬, 等.三种云南产鲜花醇提液对ABTS自由基的清除作用[J].化学与生物工程, 2019, 36(2):47-50.
NA J, MA J, ZHANG H F, et al.Scavenging effect of ethanol extract from three kinds of Yunnan flowers towards ABTS free radical[J].Chemistry & Bioengineering, 2019, 36(2):47-50.
[11] JING X L, YANG C, ZHANG L P. Characterization and analysis of protein structures in oat bran[J]. Journal of Food Science, 2016, 81(10): C2337-C2343.
[12] 冯雅蓉, 马俪珍, 李黎.SephadexG-25对羊骨胶原降血压肽的分离纯化效果研究[J].肉类研究, 2011, 25(6):30-33.
FENG Y R, MA L Z, LI L.Sephadexg-25 column chromatographic separation, MALDI-TOF-MS analysis and amino acid composition of sheep bone collagen-derived ACE inhibitory peptides[J].Meat Research, 2011, 25(6):30-33.
[13] 沈同. 生物化学(第二版)[M].北京:高等教育出版社, 1991:163-175.
SHEN T.Biochemistry (2nd Edition)[M].Beijing:Higher Education Press, 1991:163-175.
[14] 陶蔚孙. 蛋白质分子基础(第二版)[M].北京:高等教育出版社, 1995:72-86.
TAO W S.Molecular Basis of Protein (2nd Edition)[M].Beijing:Higher Education Press, 1995:72-86.
[15] 魏连会, 董艳, 石杰, 等.汉麻籽抗氧化肽的制备与氨基酸序列分析[J].中国油脂, 2022, 47(4):36-40.
WEI L H, DONG Y, SHI J, et al.Preparation and amino acid sequence analysis of antioxidant peptides from hemp seeds[J].China Oils and Fats, 2022, 47(4):36-40.
[16] 吴宝森, 王桂瑛, 谷大海, 等.诺邓火腿抗氧化肽的分离纯化与鉴定[J].食品与发酵工业, 2018, 44(7):102-109.
WU B S, WANG G Y, GU D H, et al.Isolation, purification and identification of antioxidant peptides of Nuodeng ham[J].Food and Fermentation Industries, 2018, 44(7):102-109.
[17] 邢路娟, 胡亚亚, 周光宏, 等.宣威火腿中粗肽的提取与抗氧化活性鉴定[J].南京农业大学学报, 2015, 38(4):661-666.
XING L J, HU Y Y, ZHOU G H, et al.The extraction and antioxidant activities of crude peptides from Xuanwei ham[J].Journal of Nanjing Agricultural University, 2015, 38(4):661-666.
[18] 吴淇, 邹亮华, 胡学一, 等.大麻二酚类似物的合成及其抗氧化性和抑菌性[J].精细化工, 2022, 39(3):584-591.
WU Q, ZOU L H, HU X Y, et al.Synthesis and antioxidant and bacteriostatic properties of cannabidiol analogs[J].Fine Chemicals, 2022, 39(3):584-591.
[19] CHEN H M, MURAMOTO K, YAMAUCHI F, et al.Antioxidative properties of histidine-containing peptides designed from peptide fragments found in the digests of a soybean protein[J].Journal of Agricultural and Food Chemistry, 1998, 46(1):49-53.
[20] HERNÁNDEZ-LEDESMA B, DÁVALOS A, BARTOLOMÉ B, et al.Preparation of antioxidant enzymatic hydrolysates from alpha-lactalbumin and beta-lactoglobulin.Identification of active peptides by HPLC-MS/MS[J].Journal of Agricultural and Food Chemistry, 2005, 53(3):588-593.
[21] 李东辉, 吴红伟, 李国峰, 等.基于质谱及分子对接探究大黄褐变机理[J].天然产物研究与开发, 2022, 34(10):1666-1677.
LI D H, WU H W, LI G F, et al.Investigation on browning mechanism of Rhei Radix et Rhizoma based on mass spectrometry and molecular docking[J].Natural Product Research and Development, 2022, 34(10):1666-1677.
