该研究以乌拉特戈壁红驼肉为研究对象,分别测定4 ℃条件贮藏0、1和3 d时驼肉糖酵解指标及相关酶的活性;进一步,采用磷酸化4D-Label free定量蛋白质组学技术,筛选不同成熟期驼肉差异磷酸化蛋白质,进而解析其潜在的功能信息,旨在阐明宰后不同成熟时期驼肉糖酵解变化规律及氧化磷酸化对糖酵解的影响机制,同时为改善驼肉品质提供理论基础。结果表明,随着宰后成熟时间的延长,驼肉pH值呈下降趋势,在第3天时降至5.65;蒸煮损失先上升后下降,而剪切力逐渐上升,成熟第3天达到了93.22 N。驼肉糖酵解潜力逐渐降低,成熟第3天时降到了(8.2±1.5) mmol/(g prot);糖酵解相关酶活性(如己糖激酶、乳酸脱氢酶活性等)随着成熟时间的延长呈下降趋势,而丙酮酸激酶活性呈上升趋势。磷酸化蛋白质组学的结果表明,在不同成熟期驼肉中共检测到1 840个磷酸化蛋白质,其中筛选到的差异蛋白质主要富集在磷酸原代谢、磷酸原的生物合成、磷酸肌酸代谢、肌动蛋白细胞骨架、三磷酸腺苷结合等功能过程中;且筛选出的差异蛋白质与糖酵解相关酶活性呈显著相关(P＜0.05),其中己糖激酶活性与磷酸丙酮酸水合酶呈极显著负相关(P＜0.01)。结果表明,驼肉宰后糖酵解进程主要受能量代谢和氧化还原过程的影响,研究结果为驼肉品质变化及控制的研究提供了数据支持及参考。
In this study, the glycolysis index and related enzyme activity of Urad Gobi red camel meat were measured at 0, 1 and 3 days of storage at 4 ℃.Meanwhile, phosphorylated 4D-Label free quantitative proteomics technology was used to investigate differentially phosphorylated proteins and analyzed their potential functional information in camel meat at different postmortem aging.Our research will elucidate the changes in glycolysis of camel meat at different postmortem aging after slaughter and the impact mechanism of oxidative phosphorylation on glycolysis.This will provide a theoretical basis for improving camel meat quality.The results showed that the pH of camel meat decreased with the extension of postmortem aging time, and decreased to 5.65 on 3rd day.The cooking loss increased first and then decreased, while the shear force gradually increased, reaching 93.22 N on the mature 3rd day.The glycolysis potential of camel meat gradually decreased with the extension of postmortem aging time, and decreased to (8.2±1.5) mmol/(g prot) at 3rd day.The activity of glycolysis-related enzymes such as hexokinase and lactate dehydrogenase decreased, however the activity of pyruvate kinase increased.The results of phosphorylated proteomics showed that a total of 1 840 phosphorylated proteins were detected in camel meat at different postmortem aging time, and the differentially expressed proteins were mainly enriched in the functional processes of phosphogen metabolism, phosphogen biosynthesis, phosphocreatine metabolism, actin cytoskeleton, and adenosine triphosphate binding.The differentially proteins were significantly correlated with the activities of glycolytic enzymes(P<0.05), and the hexokinase activity was significantly negatively correlated with phosphopyruvate hydratase (P<0.01).The above results showed that the glycolysis process of camel meat after slaughter was mainly affected by energy metabolism and redox process, which provided data support and reference for the study of camel meat quality change and control.
[1] 李秀丽, 双全, 乌云, 等.阿拉善双峰骆驼肉品质分析[J].食品科技, 2012, 37(7):120-123.
LI X L, SHUANG Q, WU Y, et al.Analysis on quality of Alxa Bactrian camel meat[J].Food Science and Technology, 2012, 37(7):120-123.
[2] 刘玥如. 不同年龄双峰驼不同部位肉品质特性的研究[D].呼和浩特:内蒙古农业大学, 2021.
LIU Y R.Study on meat quality characteristics of different parts of Bactrian camels at different ages[D].Hohhot:Inner Mongolia Agricultural University, 2021.
[3] KADIM I T, MAHGOUB O, PURCHAS R W.A review of the growth, and of the carcass and meat quality characteristics of the one-humped camel (Camelus dromedaries)[J].Meat Science, 2008, 80(3):555-569.
[4] 李琳. 驼肉宰后成熟过程中营养品质与肌原纤维蛋白特性的变化研究[D].呼和浩特:内蒙古农业大学, 2023.
LI L.Changes of nutritional quality and myofibrillin properties of camel meat during postmortem aging[D].Hohhot:Inner Mongolia Agricultural University, 2023.
[5] 杨丽, 傅樱花, 张兆肖, 等.骆驼肉的营养价值、食用品质及加工现状[J].肉类研究, 2018, 32(6):55-60.
YANG L, FU Y H, ZHANG Z X, et al.Current status of nutritional value, meat quality and processing of camel meat[J].Meat Research, 2018, 32(6):55-60.
[6] MOOKERJEE S A, NICHOLLS D G, BRAND M D.Determining maximum glycolytic capacity using extracellular flux measurements[J].PLoS One, 2016, 11(3):e0152016.
[7] CHAUHAN S S, ENGLAND E M.Postmortem glycolysis and glycogenolysis:Insights from species comparisons[J].Meat Science, 2018, 144:118-126.
[8] 杨媛媛, 李敬, 赵青余, 等.猪肌肉糖原酵解潜力的影响因素及其营养调控研究进展[J].中国畜牧杂志, 2019, 55(11):8-15.
YANG Y Y, LI J, ZHAO Q Y, et al.Factors affecting the glycolysis potential of pig muscle and its nutritional regulation[J].Chinese Journal of Animal Science, 2019, 55(11):8-15.
[9] 冯跃进, 顾宪红.热应激对猪肉品质的影响及其机制的研究进展[J].中国畜牧兽医, 2013, 40(2):96-99.
FENG Y J, GU X H.Research progress on the effect of heat stress on meat quality and its mechanism in pigs[J].China Animal Husbandry & Veterinary Medicine, 2013, 40(2):96-99.
[10] HUANG H G, LARSEN M R, KARLSSON A H, et al.Gel-based phosphoproteomics analysis of sarcoplasmic proteins in postmortem porcine muscle with pH decline rate and time differences[J].Proteomics, 2011, 11(20):4063-4076.
[11] 摆玉蔷. 蛋白质磷酸化对肉羊宰后糖原磷酸化酶活性的调节机制[D].北京:中国农业科学院, 2020.
BAI Y Q.The regulation mechanism of protein phosphorylation on activity of glycogen phosphorylase in postmortem lamb meat [D].Beijing:Chinese Academy of Agricultural Sciences, 2020.
[12] 刘满顺. 蛋白质磷酸化修饰对羊肉品质影响研究[D].西安:陕西师范大学, 2017.
LIU M S.THE effect of protein phosphorylation onthe ovine meat quality[D].Xi’an:Shaanxi Normal University, 2017.
[13] 陈立娟, 李欣, 杨扬, 等.不同嫩度羊肉肌浆蛋白质磷酸化水平随宰后成熟时间变化的研究[J].现代食品科技, 2015, 31(4):95-101.
CHEN L J, LI X, YANG Y, et al.Analyzing the changes in sarcoplasmic protein phosphorylation with respect to postmortem ageing times in mutton with different levels of tenderness[J].Modern Food Science and Technology, 2015, 31(4):95-101.
[14] CHEN L J, LI X, NI N, et al.Phosphorylation of myofibrillar proteins in post-mortem ovine muscle with different tenderness[J].Journal of the Science of Food and Agriculture, 2016, 96(5):1474-1483.
[15] HUANG H G, LARSEN M R, PALMISANO G, et al.Quantitative phosphoproteomic analysis of porcine muscle within 24 h postmortem[J].Journal of Proteomics, 2014, 106:125-139.
[16] 李艳娇. 日粮淀粉水平与类型对育肥猪生长和肉品质的影响及其作用机制研究[D].南京:南京农业大学, 2016.
LI Y J.Effects and mechanisms of dietary starch levels and types on growth and meat quality of finishing pigs[D].Nanjing:Nanjing Agricultural University, 2016.
[17] MONIN G, SELLIER P.Pork of low technological quality with a normal rate of muscle pH fall in the immediate post-mortem period:The case of the Hampshire breed[J].Meat Science, 1985, 13(1):49-63.
[18] WIŚNIEWSKI J R, ZOUGMAN A, NAGARAJ N, et al.Universal sample preparation method for proteome analysis[J].Nature Methods, 2009, 6(5):359-362.
[19] ENGLAND E M, MATARNEH S K, SCHEFFLER T L, et al.Altered AMP deaminase activity may extend postmortem glycolysis[J].Meat Science, 2015, 102:8-14.
[20] GRAHAME HARDIE D.Energy sensing by the AMP-activated protein kinase and its effects on muscle metabolism[J].Proceedings of the Nutrition Society, 2011, 70(1):92-99.
[21] 郑晓寒. 不同极限pH值牛肉宰后初期糖代谢分析[D].泰安:山东农业大学, 2016.
ZHENG X H.Glycolysis analysis of different ultimate pH beef during early postmortem time[D].Taian:Shandong Agricultural University, 2016.
[22] MALHEIROS J M, BRAGA C P, GROVE R A, et al.Influence of oxidative damage to proteins on meat tenderness using a proteomics approach[J].Meat Science, 2019, 148:64-71.
[23] GAGAOUA M, TROY D, MULLEN A M.The extent and rate of the appearance of the major 110 and 30 kDa proteolytic fragments during post-mortem aging of beef depend on the glycolysing rate of the muscle and aging time:An LC-MS/MS approach to decipher their proteome and associated pathways[J].Journal of Agricultural and Food Chemistry, 2021, 69(1):602-614.
[24] BELDARRAIN L R, ALDAI N, PICARD B, et al.Use of liquid isoelectric focusing (OFFGEL) on the discovery of meat tenderness biomarkers[J].Journal of Proteomics, 2018, 183:25-33.
