Melody J L, Lonergan S M, Rowe L J, Huiatt T W, Mayes M S, Huff-Lonergan E
Department of Animal Science, Iowa State University, Ames 50011, USA.
J Anim Sci. 2004 Apr;82(4):1195-205. doi: 10.2527/2004.8241195x.
The objective of this study was to determine whether differences in pork tenderness and water-holding capacity could be explained by factors influencing calpain activity and proteolysis. Halothane-negative (HAL-1843 normal) Duroc pigs (n = 16) were slaughtered, and temperature and pH of the longissimus dorsi (LD), semimembranosus (SM), and psoas major (PM) were measured at 30 and 45 min and 1, 6, 12, and 24 h postmortem. Calpastatin activity; mu-calpain activity; and autolysis and proteolysis of titin, nebulin, desmin, and troponin-T were determined on muscle samples from the LD, SM, and PM at early times postmortem. Myofibrils from each muscle were purified to assess myofibril-bound (mu-calpain. Percentage drip loss was determined, and Warner-Bratzler shear (WBS) force was analyzed. Myosin heavy-chain (MHC) isoforms were examined using SDS-PAGE. The pH of PM was lower (P < 0.01) than the pH of LD and SM at 30 and 45 min and 1 h postmortem. The PM had a higher (P < 0.01) percentage of the MHC type IIa/IIx isoforms than the LD. The-LD had the greatest proportion of (P < 0.01) MHC IIb isoforms of any of the muscles. The PM had the lowest (P < 0.01) percentage of MHC IIb isoforms and a greater (P < 0.05) percentage of type I MHC isoforms than the LD and SM. The PM had less (P < 0.01) drip loss after 96 h of storage than the SM and LD. The PM had more desmin degradation (P < 0.01) than the LD and SM at 45 min and 6 h postmortem. Degradation of titin occurred earlier in the PM than the LD and SM. At 45 min postmortem, the PM consistently had some autolysis of mu-calpain, whereas the LD and SM did not. At 6 h postmortem, some autolysis of mu-calpain (80-kDa subunit) was observed in all three muscles. The rapid pH decline and increased rate of autolysis in the PM paralleled an earlier appearance of myofibril-bound mu-calpain. The SM had higher calpastatin activity (P < 0.05) at 45 min, 6 h, and 24 h and had higher WBS values at 48 h (P < 0.01) and 120 h (P < 0.05) postmortem than the LD. At 48 and 120 h postmortem, more degradation of desmin, titin, and nebulin were observed in the LD than in the SM. These results show that mu-calpain activity, mu-calpain autolysis, and protein degradation are associated with differences in pork tenderness and water-holding capacity observed in different muscles.
本研究的目的是确定猪肉嫩度和持水能力的差异是否可以由影响钙蛋白酶活性和蛋白水解的因素来解释。屠宰了氟烷阴性(HAL - 1843正常)的杜洛克猪(n = 16),并在宰后30分钟、45分钟以及1、6、12和24小时测量背最长肌(LD)、半膜肌(SM)和腰大肌(PM)的温度和pH值。在宰后早期,测定了钙蛋白酶抑制蛋白活性、μ-钙蛋白酶活性以及肌联蛋白、伴肌动蛋白、结蛋白和肌钙蛋白-T的自溶和蛋白水解情况。从LD、SM和PM的肌肉样本中纯化肌原纤维,以评估与肌原纤维结合的(μ-钙蛋白酶)。测定了滴水损失百分比,并分析了沃纳-布拉茨勒剪切(WBS)力。使用SDS-PAGE检测肌球蛋白重链(MHC)亚型。宰后30分钟、45分钟和1小时时,PM的pH值低于LD和SM(P < 0.01)。与LD相比,PM中MHC IIa/IIx亚型的百分比更高(P < 0.01)。在所有肌肉中,LD的MHC IIb亚型比例最大(P < 0.01)。与LD和SM相比,PM的MHC IIb亚型百分比最低(P < 0.01),I型MHC亚型百分比更高(P < 0.05)。储存96小时后,PM的滴水损失比SM和LD少(P < 0.01)。宰后45分钟和6小时时,PM的结蛋白降解比LD和SM更多(P < 0.01)。肌联蛋白的降解在PM中比在LD和SM中发生得更早。宰后45分钟时,PM中μ-钙蛋白酶始终存在一些自溶现象,而LD和SM则没有。宰后6小时时,在所有三块肌肉中都观察到了μ-钙蛋白酶(80-kDa亚基)的一些自溶现象。PM中pH值的快速下降和自溶速率的增加与肌原纤维结合的μ-钙蛋白酶的更早出现相平行。SM在宰后45分钟、6小时和24小时时的钙蛋白酶抑制蛋白活性更高(P < 0.05),在宰后48小时(P < 0.01)和120小时(P < 0.05)时的WBS值更高。宰后48小时和120小时时,LD中结蛋白、肌联蛋白和伴肌动蛋白的降解比SM中更多。这些结果表明,μ-钙蛋白酶活性、μ-钙蛋白酶自溶和蛋白质降解与不同肌肉中观察到的猪肉嫩度和持水能力差异有关。