该研究对不同时期干腌牛肉进行了抗氧化和抑菌活性的评价,并发现了5个具有良好抗氧化活性的短肽。盐酸提取法发现腌制后粗肽含量相比未腌制增加了17.8%。3种不同处理方式(未处理、胃蛋白酶和胰蛋白酶)后粗肽液氨基酸含量分别为29.81、64.199、258.111 mg/mL。圆二色谱发现粗肽在195、215 nm附近有吸收带,在205 nm附近有较弱吸收带。茚三酮法测试肽链平均分子质量为19.26~27.02 kDa。通过检测DPPH自由基、超氧阴离子自由基、ABTS阳离子自由基和羟自由基清除率确定其抗氧化能力。结果表明,干腌牛肉具有较好的抗氧化活性,粗肽(1 mg/mL)对4种自由基的清除率分别为78.33%、81.05%、26.89%和36.92%。通过稳定性测试,发现0% NaCl、60 ℃和pH 7.0时抗氧化能力最强。粗肽也对O517型大肠杆菌、金黄色葡萄球菌和白色念珠菌有抑菌能力,5 mg/mL时分别可以达到75.45%、48.53%和44%。经过SephadexG-25和SephadexG-15分别进行2次分离纯化后发现5个肽具有良好的抗氧化活性,分别为DALKKK、FDGDF、HLPSDF、FLSDH、TPTDWK。所以,干腌牛肉制品可能在抗氧化以及抑菌方面促进人体健康。
This study evaluated the antioxidant and antibacterial activity of dry-cured beef at different periods and identified five short peptides with good antioxidant activity. The crude peptide content was found to increase by 17.8% after curing compared to uncured by hydrochloric acid extraction method. The amino acid contents of crude peptide solution after three different treatments (untreated, pepsin, and trypsin) were 29.81 mg/mL, 64.199 mg/mL, and 258.111 mg/mL, respectively. The circular dichroism spectrum showed that crude peptides had absorption bands around 195 nm and 215 nm and a weak absorption band around 205 nm. The average molecular weight distribution of the peptide chains tested by the ninhydrin method ranged from 19.26 kDa to 27.02 kDa. The antioxidant capacity was determined by detecting the scavenging rates of DPPH free radicals, superoxide anion radicals, ABTS cationic radicals, and hydroxyl radicals. Results showed that the dry-cured beef had good antioxidant activity, and the scavenging rates of the crude peptide (1 mg/mL) were 78.33%, 81.05%, 26.89%, and 36.92% for the four free radicals, respectively. By stability test, the antioxidant capacity was strongest at 0% NaCl, 60 ℃, and pH 7.0. The crude peptides also had antibacterial ability against Escherichia coli O517, Staphylococcus aureus, and Candida albicans, which could reach 75.45%, 48.53%, and 44% at 5 mg/mL, respectively. After two isolations and purifications by SephadexG-25 and SephadexG-15, respectively, five peptides were found to have good antioxidant activity, namely DALKKK, FDGDF, HLPSDF, FLSDH, and TPTDWK. Therefore, dry corned beef products may promote human health in terms of antioxidant as well as bacterial inhibition.
[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 oil fermented meat products[J].Food Science, 2011, 32(3):247-251.
[2] 马菊, 孙宝忠, 郝永清.国内外发酵肉制品历史及发展现状比较[J].肉类研究, 2006, 20(9):45-47.
MA J, SUN B Z, HAO Y Q.Comparisons with history and the current status of fermented meat productions at home and abroad[J].Meat Research, 2006, 20(9):45-47.
[3] BHAT Z F, KUMAR S, BHAT H F.Bioactive peptides of animal origin:A review[J].Journal of Food Science and Technology, 2015, 52(9):5377-5392.
[4] 胡亚亚, 邢路娟, 周光宏, 等.不同提取方法对金华火腿粗肽液抗氧化活性的影响[J].食品工业科技, 2015, 36(14):115-118.
HU Y Y, XING L J, ZHOU G H, et al.Effect of extraction methods on the antioxidant activity of crude peptides from Jinhua ham[J].Science and Technology of Food Industry, 2015, 36(14):115-118.
[5] 左庆翔. 金华火腿降血压肽的特性研究及分离纯化[D].南京:南京农业大学, 2017.
ZUO Q X.Study on the characteristics of Jinhua ham hypotensive peptide and its isolation and purification[D].Nanjing:Nanjing Agricultural University, 2017.
[6] 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.
[7] 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 Nordeng ham[J].Food and Fermentation Industries, 2018, 44(7):102-109.
[8] ZHU C Z, ZHANG W G, KANG Z L, et al.Stability of an antioxidant peptide extracted from Jinhua ham[J].Meat Science, 2014, 96(2):783-789.
[9] 徐娟, 吕嘉枥.乳蛋白水解液中多肽含量测定方法的研究[J].食品科技, 2010, 35(12):275-278.
XU J, LYU J F.Determination of content of peptides in milk protein hydrolysates[J].Food Science and Technology, 2010, 35(12):275-278.
[10] 严群芳. 大豆抗氧化肽的分离及其生物活性研究[D].南京:南京农业大学, 2006.
YAN Q F.Study on the separation of soybean antioxidative peptides and their antioxidant activity[D].Nanjing:Nanjing Agricultural University, 2006.
[11] LI Y H, JIANG B, ZHANG T, et al.Antioxidant and free radical-scavenging activities of chickpea protein hydrolysate (CPH)[J].Food Chemistry, 2008, 106(2):444-450.
[12] LIU F, OOI V E, CHANG S T.Free radical scavenging activities of mushroom polysaccharide extracts[J].Life Sciences, 1997, 60(10):763-771.
[13] VISESSANGUAN W, BENJAKUL S, RIEBROY S, et al.Changes in composition and functional properties of proteins and their contributions to Nham characteristics[J].Meat Science, 2004, 66(3):579-588.
[14] BRUNTON N P, LYNG J G, ZHANG L, et al.The use of dielectric properties and other physical analyses for assessing protein denaturation in beef biceps femoris muscle during cooking from 5 to 85 ℃[J].Meat Science, 2006, 72(2):236-244.
[15] GALLEGO M, GROOTAERT C, MORA L, et al.Transepithelial transport of dry-cured ham peptides with ACE inhibitory activity through a Caco-2 cell monolayer[J].Journal of Functional Foods, 2016, 21:388-395.
[16] LI B B, XU Y, LI J, et al.Effect of oxidized lipids stored under different temperatures on muscle protein oxidation in Sichuan-style sausages during ripening[J].Meat Science, 2019, 147:144-154.
[17] CHEN L L, SONG L Y, LI T F, et al.A new antiproliferative and antioxidant peptide isolated from Arca subcrenata[J].Marine Drugs, 2013, 11(6):1800-1814.
[18] CHAN W K M, DECKER E A, LEE J B, et al.EPR spin-trapping studies of the hydroxyl radical scavenging activity of carnosine and related dipeptides[J].Journal of Agricultural and Food Chemistry, 1994, 42(7):1407-1410.
[19] CASTELLANO P, MORA L, ESCUDERO E, et al.Antilisterial peptides from Spanish dry-cured hams:Purification and identification[J].Food Microbiology, 2016, 59:133-141.
[20] VASTAG Z, POPOVIC L, POPOVIC S, et al.Antioxidant and angiotensin-I converting enzyme inhibitory activity in the water-soluble protein extract from Petrovac Sausage (Petrovská Kolbása)[J].Food Control, 2010, 21(9):1298-1302.
[21] KĘSKA P, WÓJCIAK K M, STADNIK J.Bioactive peptides from beef products fermented by acid whey-in vitro and in silico study[J].Scientia Agricola, 2019, 76(4):311-320.
[22] GALLEGO M, MORA L, ESCUDERO E, et al.Bioactive peptides and free amino acids profiles in different types of European dry-fermented sausages[J].International Journal of Food Microbiology, 2018, 276:71-78.
