20190806 周二
体外氧化对牛肉肌原纤维蛋白结构和功能的影响
J. Agric. Food Chem. (IF: 3.4，1区)
推荐|莫玲
研究背景
鲜肉肌原纤维蛋白分离物(MPI)对活性氧引发的氧化反应高度敏感。当ROS在生理条件下超过内源性抗氧化防御时，ROS导致蛋白质氧化。牛MPI的氧化可引起蛋白质结构的改变、肽骨架的裂解、氨基酸侧链的修饰以及蛋白质分子间交联形成引起的聚集。氧化引起的最显著的后果主要包括蛋白质羰基衍生物的积累、游离巯基的减少、表面疏水性的增加、蛋白质的结构展开和蛋白质交联的形成。蛋白质氧化修饰、脂质氧化和微生物腐败是鲜肉加工、加工和贮藏过程中鲜肉食用品质下降的关键因素。蛋白质氧化还能引起人体氧化应激，导致许多疾病，如帕金森病、阿尔茨海默病和白内障。嫩度被牛肉行业视为牛肉品质的关键，它受到多种因素的影响。在所有因素中，关键结构蛋白的降解速度和降解程度对提高死后牛肉嫩度至关重要。钙蛋白酶系统,特别是μ-钙蛋白酶,已发现早期死亡蛋白水解作用调节中发挥关键作用的牛肉蛋白质与嫩度。以前的研究表明，氧化会对μ-钙蛋白酶的活性和自溶作用产生负面影响，这主要调节关键肌原纤维蛋白的降解，最终影响牛肉的嫩度。然而，关于氧化如何影响MPI的结构和生化性质的潜在机制尚未研究。因此，本研究的主要目的是检测蛋白质氧化对来自牛肉肌肉的肌原纤维蛋白质的生化特性的影响，并进一步使用蛋白质组学技术鉴定易受影响的蛋白质进行氧化反应。
研究方法
1、肌原纤维蛋白的氧化：肌纤维蛋白(20mg/mL)与三氯化铁(FeCl3),和不同浓度的过氧化氢在4℃条件下反应24 h，加入EDTA停止反应。
2、羰基含量测定
3、游离巯基含量
4、表面疏水性
5、颗粒直径
6、SDS-PAGE和Western blot 分析
7、LC-MS/MS法鉴定氧化蛋白
研究结果
1、氧化引起了牛MPI结构的剧烈变化。随着过氧化氢浓度的增加，MPI的羰基含量、表面疏水性和粒径显著增加，而游离巯基含量显著降低。
2、氧化作用通过蛋白质和氨基酸的交联使蛋白质聚集。
3、蛋白质组学研究确定了易被氧化的蛋白质位点。分别鉴定了易氧化酶的氧化催化活性和结合位点
研究结论
本研究提供证据表明，蛋白质氧化可能参与了死后陈化过程中鲜肉品质的调控。
原文链接
Effects of oxidation in vitro on structures and functions of myofibrillar protein from beef muscles. by Qingquan Fu, Rui Liu, …, Wangang Zhang (2019)
https://pubs.acs.org/doi/10.1021/acs.jafc.9b01239
学者简介
张万刚
张万刚，教授，博士生导师，本科和硕士毕业于中国农业大学，博士和博士后毕业于美国爱荷华州立大学，食品科学与工程系系主任，现担任国际 SCI 期刊《Meat Science》副主编、中文核心期刊《中国畜牧杂志》副主编、中国畜产品加工研究会肉品加工专业委员会副主任委员兼秘书长。
科研情况：
在研项目：
国家自然基金面上项目， 蛋白质亚硝基化对牛肉成熟嫩化过程中细胞
凋亡的调控机理研究，31871827，2019.1-2022.12，60 万；
国家自然基金面上项目， 蛋白质亚硝基化对猪肉持水力的作用机理研
究，31571853，2016.1-2019.12，75 万；
“十三五”国家重点研发计划课题， 禽畜水产类方便即食食品制造关键
技术开发研究及新产品创制， 2016YFD0400703， 2016.7-2020.12， 500
万；
“十三五”国家重点研发计划子课题，民族特色工业化肉制品加工关
键技术与装备开发，2018YFD0400101，2018.7-2020.12，68 万；
中央基本科研业务费，一氧化氮调控猪肉滴水损失的机理研究，
KYDZ201902，2019.1- 2020.12，20 万;
中央基本科研业务费，多浪羊宰后品质控制与休闲羊肉食品开发，
KYLH201701，2017.1- 2019.12，20 万。
发表学术论文（第一作者或通讯作者）：
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3. Liu, R., Zhang, C. Y., Xing, L. J., Zhang, L. L., Zhou, G. H., & Zhang, W. G.* (2019). A bioinformatics study on characteristics, metabolic pathways, and cellular functions of the
identified S-nitrosylated proteins in postmortem pork muscle. Food Chemistry, 274, 407-414.
4. Xiao, Z. C., Ge, C. R., Zhou, G. H., Zhang, W. G.*, & Liao, G. Z. (2019).1H NMR-based metabolic characterization of Chinese Wuding Chicken meat. Food Chemistry, 274, 574-582.
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6. Liu, R., Lonergan, S., Steadham, E., Zhou, G. H., Zhang, W. G.*, & Huff-Lonergan, E.(2019). Effect of nitric oxide on myofibrillar proteins and the susceptibility to calpain-1
proteolysis. Food Chemistry, 276, 63-70.
7. Xing, L. J., Ge, Q. F., Jiang, D. L., Gao, X. G., Liu, R., Cao, S. M., Zhuang, X. B., Zhou, G.H., & Zhang, W. G.* (2018). Caco-2 cell-based electrochemical biosensor for evaluating the antioxidant capacity of Asp-Leu-Glu-Glu isolated from dry-cured Xuanwei ham. Biosensors and Bioelectronics, 105, 81-89.
8. Ge, Q. F., Ge, P. W., Du, N., Chen, J. H., Yuan, L. M., Yu, H., Xu, X., Wu, M. G., Zhang, W. G.*, & Zhou, G. H. (2018). A novel and simple cell-based electrochemical biosensor for evaluating the antioxidant capacity of Lactobacillus plantarum strains isolated from Chinese dry-cured ham. Biosensors and Bioelectronics, 99, 555-563.
9. Zou, Y. H., Liu, R., Qi, J., Zhou, G. H., & Zhang, W. G.* (2018). Effects of ultrasonic
assisted cooking on the chemical profiles of taste and flavor of spiced beef. Ultrasonics Sonochemistry, 46, 36-45.
10. Xing, L. J., Liu, R., Gao, X. G., Zheng, J. X., Wang, C., Zhou, G. H., & Zhang, W. G.* (2018). The proteomics homology of antioxidant peptides extracted from dry-cured Xuanwei and Jinhua ham. Food Chemistry, 266, 420-426.
11. Wang, A. R., Kang, D. C., Zhang, W. G.*, Zhang, C. Y., Zou, Y. H., & Zhou, G. H. (2018). Changes in calpain activity, protein degradation and microstructure of beef M.
semitendinosus by the application of ultrasound. Food Chemistry, 2018, 245, 724-730.
12. Liu, R., Warner, R., Zhou, G. H., & Zhang, W. G.* (2018). Contribution of nitric oxide and protein S-nitrosylation to variation in fresh meat quality. Meat Science, 144, 135-148.
13. Liu, R., Fu, Q. Q., Lonergan, S., Huff-Lonergan, E., Xing, L. J., Zhang, L. L., Bai, Y., Zhou,G. H., & Zhang, W. G.* (2018). Identification of S-nitrosylated proteins in postmortem pork muscle using modified biotin switch method coupled with isobaric tags. Meat Science, 145,431-439.
14. Zhang, C. Y., Liu, R., Wang, A. R., Kang, D. C., Zhou, G. H., & Zhang, W. G.* (2018).Regulation of calpain-1 activity and protein proteolysis by protein nitrosylation in
postmortem beef. Meat Science, 141, 44-49.
15. Zhao, Y. Y, Hou, Q., Zhuang, X. B., Wang, Y., Zhou, G. H., & Zhang, W. G.* (2018). Effect of regenerated cellulose fiber on the physicochemical properties and sensory characteristics of fat-reduced emulsified sausage. LWT-Food Science and Technology, 97, 157-163.
16. Xiao, Z. C., Luo, Y. T., Wang, G. Y., Ge, C. R., Zhou, G. H., Zhang, W. G.*, & Liao, G. Z.(2018).1H NMR-based water-soluble lower molecule characterization and fatty acid
composition of boiled Wuding Chicken during processing. Journal of the Science of Food
andAgriculture,Accepted.
17. Zou, Y. H., Zhang, W. G.*, Kang, D. C., & Zhou, G. H. (2018). Improvement of tenderness and water holding capacity of spiced beef by the application of ultrasound during cooking.International Journal of Food Science and Technology, 53, 826-836.
18. Xing, L. J., Liu, R., Tang, C. B., Pereira, J., Zhou, G. H., and Zhang, W. G.* (2018). The antioxidant activity and transcellular pathway of Asp-Leu-Glu-Glu in a Caco-2 cell monolayer. International Journal of Food Science and Technology, 53, 2405- 2414.
19. Kang, D. C., Gao, X. Q., Ge, Q. F., Zhou, G. H., and Zhang, W. G.* (2017). Effects of ultrasound on the beef structure and water distribution during curing through protein degradation and modification. Ultrasonics Sonochemistry, 38, 317-325.
20. Zhang, W. G., Naveena, B. M., Jo, C., Sakata, R., Zhou, G. H., Banerjee, R., and Nishiumi,T. (2017). Technological demands of meat processing-An Asian perspective. Meat Science, 132, 35-44.
21. Kang, D. C., Jiang, Y. H., Xing, L. J., Zhou, G. H., and Zhang, W. G.* (2017). Inactivation of Escherichia coli O157:H7 and Bacillus cereus by power ultrasound during the curing processing in brining liquid and beef. Food Research International, 102, 717-727.
22. Fu, Q. Q., Ge, Q. F., Liu, R., Wang, H. O., Zhou, G. H., and Zhang, W. G.* (2017).
Influence of modified atmosphere packaging on protein oxidation, calpain activation and desmin degradation of beef muscles. Journal of the Science of Food and Agriculture, 97,4508-4514.
23. Hu, H. Y., Pereira, J., Xing, L. J., Zhou, G. H., and Zhang, W. G.* (2017). Thermal gelation and microstructural properties of myofibrillar protein gel with the incorporation of
regenerated cellulose. LWT-Food Science and Technology, 86, 14-19.
24. Ge, Q. F., Gu, Y. B., Zhang, W. G.*, Yin, Y. Q., Yu, H., Wu, M. G., Wang, Z. J., and Zhou, G.H. (2017). Comparison of microbial communities from different Jinhua ham factories. AMB Express, 7, DOI: 10.1186/s13568-017-0334-0.
25. Fu, Q. Q., Liu, R., Zhou, G. H., and Zhang, W. G.* (2017). Effects of packaging methods on the color of beef muscles through influencing myoglobin status, metmyoglobin reductase activity and lipid oxidation. Journal of Food Processing and Preservation, 41, e12740, DOI:10.1111/jfpp.12740.
26. Liu R., Xing L. J., Zhou G. H., and Zhang, W. G.* (2017). What is meat in China? Animal Frontiers, 2017, 7, 53-56.
27. Kang, D. C., Zou, Y. H., Cheng, Y. P., Xing, L. J., Zhou, G. H., and Zhang, W. G.* (2016).Effects of power ultrasound on oxidation and structure of beef proteins during curing.Ultrasonics Sonochemistry, 33, 47-53.
28. Hu, H. Y., Xing, L. J., Hu, Y. Y., Qiao, C. L., Wu, T., Zhou, G. H., and Zhang, W. G.* (2016).Effects of regenerated cellulose on oil-in-water emulsions stabilized by sodium caseinate. Food Hydrocolloids, 52, 38-46.
29. Xing, L. J., Hu, Y. Y., Hu, H. Y., Ge, Q. F., Zhou, G. H., and Zhang, W. G. * (2016).
Purification and identification of antioxidative peptides from dry-cured Xuanwei ham. Food
Chemistry, 194, 951-958.
30. Liu, R., Li, Y. P., Wang, M. Q., Zhou, G. H., and Zhang, W. G.* (2016). Effect of protein S-nitrosylation on autolysis and catalytic ability of μ-calpain. Food Chemistry, 213, 470-477.
31. Kang, D. C., Wang, A. R., Zhou, G. H., Zhang, W. G.*, Xu, S. M., and Guo, G. P. (2016).Power ultrasonic on mass transport of beef: effects of ultrasound intensity and NaCl concentration. Innovative Food Science and Emerging Technologies, 35, 36-44.
32. Hu, H. Y., Pereira, J., Xing, L. J., Hu, Y. Y., Qiao, C. L., Zhou, G. H., and Zhang, W. G.*(2016). Effects of regenerated cellulose emulsion on the quality of emulsified sausage.LWT-Food Science and Technology, 70, 315-321.
33. Gao, X. Q., Hao, X. Z., Xiong, G. H., Ge, Q. F., Zhang, W. G.*, Zhou, G. H., and Yue, X. B.(2016). Interaction between carrageenan/soy protein isolates and salt-soluble meat protein.Food and Bioproducts Processing, 100, 47-53.
34. Yin, Y., Xing, L. J., Zhou, G. H., and Zhang, W. G.* (2016). Antioxidative and antibacterial activities of rosemary extract in raw ground pork patties. Journal of Food and Nutrition Research, 4, 806-813.
35. Jailson, P., Zhou, G. H., and Zhang, W. G.* (2016). Effects of rice flour on emulsion stability,organoleptic characteristics and thermal rheology of emulsified sausage. Journal of Food and Nutrition Research, 4, 216-222.
36. Hu, Y. Y., Xing, L. J., Zhou, G. H., and Zhang, W. G.* (2016). Antioxidant activity of crude peptides extracted from dry-cured Jinhua ham. Journal of Food and Nutrition Research, 4,377-387.
37. Wang, J., Yan, X. L., Liu, R., Fu, Q. Q., Zhou, G. H., and Zhang, W. G.* (2016). Differences in calpain system, desmin degradation and water holding capacity between commercial Meishan and Duroc×Landrace×Yorkshire crossbred pork. Animal Science Journal, 87,109–116.
38. Liu, R., Xing, L. J., Fu, Q. Q., Zhou, G. H., and Zhang, W. G.* (2016). A review of
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40. Liu, R., Li, Y. P., Zhang, W. G.*, Fu, Q. Q., Liu, N., and Zhou, G. H. (2015). Activity and expression of nitric oxide synthase in pork skeletal muscles. Meat Science, 99, 25-31.
41. Fu, Q. Q., Liu, R., Zhang, W. G.*, Li, Y. P., Wang, J., and Zhou, G. H. (2015). Effects of different packaging systems on beef tenderness through protein modifications. Food and Bioprocess Technology, 8, 580-588.
42. Gao, X. Q., Kang, Z. L., Zhang, W. G.*, Li, Y. P., and Zhou, G. H. (2015). Combination of κ-carrageenan and soy protein isolates effects on functional properties of chopped low fat pork batters during heat-induced gelation. Food and Bioprocess Technology, 8, 1524-1531.
43. Gao, X. Q., Zhang, W. G.*, and Zhou, G. H. (2015). Emulsion stability, thermo-rheology and quality characteristics of ground pork patties prepared with soy protein isolate and carrageenan. Journal of the Science of Food andAgriculture, 95, 2832-2837.
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46. Gao, X. Q., Zhang, W. G.*, and Zhou, G. H. (2014). Effects of glutinous rice flour on the physiochemical and sensory qualities of ground pork patties. LWT-Food Science and Technology, 58, 135-141.
47. Yin, Y., Zhang, W. G.*, Zhou, G. H., and Guo, B. (2014). Comparison of protein degradation,protein oxidation, and µ-calpain activation between pale, soft, exudative and red, firm, non-exudative pork during postmortem aging. Journal ofAnimal Science, 92, 3745-3752.
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科研成果：
先后主持国家自科学基金面上项目、“十二五”国家科技支撑计划课题、“十三五”国家重点研发计划课题、国家回国人员择优资助项目等项目。主编英文著作《Spoilage Microorganisms in Food》，主编中文著作《冷却猪肉加工技术》，获大北农青年学者奖，以主要完成人获神农中华农业奖优秀创新团队奖和中国食品科学技术学会技术创新奖一等奖，申请或授权发明专利 14 项
http://food.njau.edu.cn/__local/5/31/C8/5ABF145560B72EC324748B27845_3F00E458_33B68.pdf https://obgyn.mcw.edu/research/staff/huang/