基于蛋白质组学分析蛋白酶体对宰后秦川牛肉贮藏期间品质变化的影响

摘要/Abstract

摘要： 摘要：本研究以宰后4 ℃条件下不同贮藏期的秦川牛背最长肌为研究对象，测定品质指标、能量水平、蛋白酶体活性变化，鉴定与蛋白酶体相关的差异蛋白质（Differential expression proteins，DEPs），利用生物信息学方法分析蛋白酶体对宰后牛肉品质及能量代谢的响应机制，结果如下：随贮藏时间的延长，pH值呈先降低后升高趋势；b*值和MFI呈上升趋势；L*值、a*值、离心损失、蒸煮损失、剪切力呈先上升后下降趋势；能量基本物质 ATP、ADP、AMP含量呈下降趋势；20S蛋白酶体活性显著降低（P < 0.05）；相关性分析结果显示：20S蛋白酶体活性与b*呈显著负相关（P < 0.05），与ADP含量呈显著正相关（P < 0.05），与AMP含量呈极显著正相关（P < 0.01），与MFI呈极显著负相关（P < 0.01），表明蛋白酶体活性与能量代谢及品质间存在密切联系。结合4D-LFQ蛋白质组学技术，鉴定出8种差异表达的蛋白酶体亚基，及27种相关DEPs；GO分析和KEGG Pathway分析发现，这些蛋白酶体亚基及其相关的DEPs具有内肽酶活性、肌动蛋白结合、微丝运动活性等分子功能，参与泛素依赖性蛋白质分解代谢、骨骼肌收缩、肌肉收缩、糖酵解等生物过程，引起秦川牛肉品质变化；宰后初期蛋白酶体通过消耗能量物质分解代谢蛋白调控生物途径，后期能量物质耗尽，蛋白酶体可通过糖酵解提供的能量以及非ATP依赖型蛋白酶体亚基，实现对宰后蛋白质的分解代谢，最终影响秦川牛肉的品质。

关键词: 关键词：秦川牛, 蛋白酶体, 4D-非标蛋白质组学, 肉品质

Abstract: Abstract：In this study, the longissimus dorsi muscle of Qinchuan cattle stored at 4 ℃ after slaughter was used as the object of study to determine quality indicators, energy levels and proteasome activity, and to identify DEPs associated with the proteasome. The results were as follows: pH decreased and then increased with increasing storage time; b* and MFI increased; L*, a*, centrifugal loss, cooking loss and shear force increased and then decreased; ATP, ADP and AMP the basic energy substances, decreased; 20S proteasome activity decreased significantly (P < 0.05); The correlation analysis showed that the 20S proteasome activity was significantly negatively correlated with b* (P < 0.05), with ADP content (P < 0.05), with AMP content (P < 0.01), and with MFI (P < 0.01), indicating a close relationship between proteasome activity and energy metabolism and beef quality. Combined with 4D-LFQ proteomics technology, eight differentially expressed proteasome subunits, and 27 associated DEPs were identified; GO analysis and KEGG Pathway analysis revealed that these proteasome subunits and their associated DEPs have molecular functions such as endopeptidase activity, actin binding, microfilament motility activity, and are involved in ubiquitin-dependent proteolytic metabolism, skeletal muscle contraction, In the early postmortem period, the proteasome regulated the biological pathway by consuming energetic material to catabolize proteins, and in the later period, when the energetic material was depleted, the proteasome could realize the catabolism of postmortem proteins through the energy provided by glycolysis and non-ATP-dependent proteasome subunits, which eventually affected the quality of Qinchuan beef.

Key words: Key words: Qinchuan beef, Proteasome, 4D-label free quantification, Meat quality

中图分类号:

TS251.53

胡丽筠王金霞李荣张倩陈雪妍李亚蕾罗瑞明. 基于蛋白质组学分析蛋白酶体对宰后秦川牛肉贮藏期间品质变化的影响[J]. 食品科学.

Li-Jun HU Jinxia Wang Xue-yan CHEN Rui-Ming LUO. Proteomics-based analysis of the effect of proteasome on quality changes in postmortem Qinchuan beef during storage[J]. FOOD SCIENCE.

参考文献

[1]黄峰, 魏起超, 李侠, 等.细胞凋亡对宰后肌肉嫩化作用机理的研究进展[J].中国农业科学, 2021, 54(10):2192-2202 [2]ALSAYYAH C, OZTURK O, CAVELLINI L, et al.The regulation of mitochondrial homeostasis by the ubiquitin proteasome system[J].Biochimica biophysica acta Bioenergetics, 2020, 1861(12):148-302 [3]DUTAUD D, AUBRY L, SENTANDREU M A, et al.Bovine muscle 20S proteasome: ISimple purification procedure and enzymatic characterization in relation with postmortem conditions[J].Meat science, 2006, 74(2):327-336 [4]TAYLOR R G, GEESINK G H, THOMPSON V F, et al.Is Z-disk degradation responsible for postmortem tenderization?[J].Journal of Animal Science, 1995, 73(5):1351-1367 [5]WIMMERS K, MURANI E, PONSUKSILI S.Functional genomics and genetical genomics approaches towards elucidating networks of genes affecting meat performance in pigs[J].Briefings in functional genomics, 2010, 9(3):251-8 [6]CHENG H, SONG S, PARK T S, et al.Comparison of Meat Quality Characteristics and Proteolysis Trends associated with Muscle Fiber Type Distribution between Duck Pectoralis Major and Iliotibialis Muscles[J].Food science of animal resources, 2022, 42(2):266-279 [7]李桂霞, 李欣, 李铮, 等.宰后僵直及成熟过程中羊背最长肌理化性质的变化[J].食品科学, 2017, 38(21):112-118 [8]马旭华.秦川牛宰后肉品质变化对HSPA6差异表达的响应机理[D]. 宁夏大学, 2022. [9]罗辉, 杨波, 李亚蕾, 等.宰后成熟期间能量物质、值和肌原纤维小片化指数对秦川牛肉嫩度的影响及其机理[J].食品科学, 2022, 43(11):171-179 [10]张杏亚, 杨波, 李亚蕾, 等.基于蛋白质组学技术研究秦川牛肉宰后贮藏过程中肌红蛋白含量及其衍生物转化[J].食品科学, 2021, 42(07):226-231 [11]高永芳.CaCl2处理对不同海拔高度牛肉宰后成熟过程中AMPK活性、能量代谢及肉质影响的研究[D]. 甘肃农业大学, 2020. [12]郑晓寒.不同极限pH值牛肉宰后初期糖代谢分析[D]. 山东农业大学, 2016. [13]杨巧能, 梁琪, 文鹏程, 等.宰后成熟时间对不同年龄牦牛肉用品质的影响[J].食品科学, 2015, 36(18):237-241 [14]明丹丹, 张一敏, 董鹏程, 等.牛肉肉色的影响因素及其控制技术研究进展[J].食品科学, 2020, 41(01):284-291 [15]刘腾.分割牛肉冷却成熟及贮藏对其品质的影响研究[D]. 福建农林大学, 2016. [16]郭荣珍, 杨锋, 刘纯友, 等.不同包装方式对冷藏过程中水牛肉品质特性的影响[J/OL]. [J]., 2022, :1-16 [17]李若绮.宰后冷藏过程中牛肉肉用品质变化及抗氧化物质对肉色稳定性影响[D]. 甘肃农业大学, 2015. [18]周荣梅, 苏伟, 母应春, 等.电磁场保鲜和冷冻贮藏对贵州黄牛肉品质的影响[J].食品与发酵科技, 2022, 58(04):14-19 [19]SMITH M A, BUSH R D, VAN DE VEN R J, et al.Effect of electrical stimulation and ageing period on alpaca (Vicugna pacos) meat and eating quality[J]. [J].Meat science, 2016, 111:38-46 [20]舒一梅, 郑翔, 达小梅, 等.不同排酸时间对凉山黄牛肉品质的影响[J].食品工业科技, 2022, 43(01):344-350 [21]魏起超.线粒体损伤对宰后牛肉嫩化的影响[D]. 中国农业科学院, 2021. DOI: 10.13386/j.issn1002-0306.2021050211. [22]MIHAYLOVA M M, SHAW R J.The AMPK signalling pathway coordinates cell growth,autophagy and metabolism[J].Nature cell biology, 2011, 13(9):1016-23 [23]ZHANG Y, ERTBJERG P.Effects of frozen-then-chilled storage on proteolytic enzyme activity and water-holding capacity of pork loin[J]. [J].Meat science, 2018, 145:375-382 [24]曾珍.钙调蛋白酶2、20S蛋白酶体和组织蛋白酶B+L对宰后猪肉保水性的影响[D]. 四川农业大学, 2018. [25]DAVIS P R, MILLER S G, VERHOEVEN N A, et al.Increased AMP deaminase activity decreases ATP content and slows protein degradation in cultured skeletal muscle[J]. [J].Metabolism, 2020, 108:154-257 [26]ABI HABIB J, LESENFANTS J, VIGNERON N, et al.Functional Differences between Proteasome Subtypes[J].Cells, 2022, 11(3):421- [27]赵宝昌, 崔秀云.蛋白酶体的结构与功能[J].生命的化学, 2002, 22(3):215-218 [28]吴潇.猪泛素—蛋白酶体途径新基因定位、序列及性状关联分析[D]. 华中农业大学动物遗传育种与繁殖, 2006. [29]MA L, LI X, ZHAO X, et al.Oxaliplatin promotes siMAD2L2 induced apoptosis in colon cancer cells[J].Molecular medicine reports, 2021, 24(3):629- [30]DU T, SONG Y, RAY A, et al.Ubiquitin receptor PSMD4/Rpn10 is a novel therapeutic target in multiple myeloma[J]. [J].Blood, 2023, :- [31]WU S, LIU H, LI S, et al.Transcriptome Analysis Reveals Possible Immunomodulatory Activity Mechanism of Chlorella spExopolysaccharides on RAW264.7 Macrophages[J].Marine drugs, 2021, 19(4):217- [32]BIRAN A, MYERS N, STEINBERGER S, et al.The C-Terminus of the PSMA3 Proteasome Subunit Preferentially Traps Intrinsically Disordered Proteins for Degradation[J].Cells, 2022, 11(20):3231- [33]ROUSSEAU A, BERTOLOTTI A.Regulation of proteasome assembly and activity in health and disease[J].Nature reviews molecular cell biology, 2018, 19(11):697-712 [34]王邵雨.泛素特异性蛋白酶8(USP8)调控湖羊肌卫星细胞增殖与分化的研究[D]. 浙江大学, 2019. [35]SHI H B, SHAHIDI F, WANG J K, et al.Techniques for postmortem tenderisation in meat processing: effectiveness, application and possible mechanisms[J]. [J].Food ProdProcess, 2021, 21:- [36]LIU Y, DU M, LI X, et al.Role of the ubiquitin-proteasome pathway on proteolytic activity in postmortem proteolysis and tenderisation of sheep skeletal muscle[J].International journal of food science and technology, 2016, 51(11):2353-2359 [37]JIA W, ZHANG R, LIU L, et al.Molecular mechanism of protein dynamic change for Hengshan goat meat during freezing storage based on high-throughput proteomics[J]. [J].Food research international, 2021, 143:110-289 [38]Cheng H, Song S, Park T S, et al.Comparison of Meat Quality Characteristics and Proteolysis Trends associated with Muscle Fiber Type Distribution between Duck Pectoralis Major and Iliotibialis Muscles[J].Food science of animal resources, 2022, 42(2):266-279 [39]C?NSOLO N R B, ROSA A F, BARBOSA L C G S, et al.Preliminary study on the characterization of Longissimus lumborum dark cutting meat in Angus × Nellore crossbreed cattle using NMR-based metabolomics[J]. [J].Meat science, 2021, 172:108-350 [40]HUANG C Y, HOU C L, IJAZ M, et al.Proteomics discovery of protein biomarkers linked to meat quality traits in post-mortem muscles: Current trends and future prospects[J].[J]. Food science & technology, 2020, 105:416-432 [41]张高伟, 吴彪, 刘志鸿, 等.魁蚶-α 基因结构特征及对低氧胁迫的响应[J].中国水产科学, 2019, 26(4):646-656 [42]陈实.捻转血矛线虫蛋白酶体β5亚基编码基因的结构与功能研究[D]. 湖北: 华中农业大学, 2014.