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1.
Pale, soft, and exudative (PSE) meat has been recognized for decades. Fast glycolysis during early post-mortem stage while the muscle temperature is still high is the cause of PSE meat. To elucidate the molecular mechanism underlying this fast glycolysis in muscle to become PSE meat, post-mortem ATP metabolism, fructose-2,6-diphosphate content, and the activities of AMPK, glycogen phosphorylase, and pyruvate kinase were examined in post-mortem muscle. Earlier and faster post-mortem AMPK activation was responsible for the significantly lower pH and higher lactic acid accumulation (p<0.05) seen in PSE muscle, which resulted in the occurrence of PSE meat. In muscle that became PSE meat, AMPK was activated at 0 h post-mortem and reached maximal activation at 0.5 h post-mortem, whereas AMPK reached maximal activation at 1 h post-mortem in the normal pork loin. Higher fructose-2,6-diphosphate content (p<0.05) was detected in PSE muscle compared to normal muscle at early post-mortem stage. However, no difference in the activities of glycogen phosphorylase and pyruvate kinase, rate-controlling enzymes in glycogenolysis and glycolysis, respectively, was detected between PSE and normal pork loins. Because fructose-2,6-diphosphate is a product of phosphofructokinase-2 (PFK-2), these data suggest that AMPK regulates post-mortem glycolysis through its phosphorylation and activation of PFK-2, which then up-regulates the activity of phosphofructokinase-1 (PFK-1), a key rate-controlling enzyme in glycolysis. Early AMPK activation in PSE muscle is associated with early consumption of ATP, because higher AMP and IMP contents and lower ATP content were detected in PSE meat compared to normal meat. Other mechanisms causing early AMPK activation in PSE meat may exist, which warrants further investigation.  相似文献   

2.
Low-field NMR T(2) and Fourier transform infrared (FT-IR) measurements were performed on meat samples of two qualities (normal and high ultimate pH) during cooking from 28 degrees C to 81 degrees C. Pronounced changes in both T(2) relaxation data and FT-IR spectroscopic data were observed during cooking, revealing severe changes in the water properties and structural organization of proteins. The FT-IR data revealed major changes in bands in the amide I region (1700-1600 cm(-)(1)), and a tentative assignment of these is discussed. Distributed NMR T(2) relaxation data and FT-IR spectra were compared by partial least-squares regression. This revealed a correlation between the FT-IR peaks reflecting beta-sheet and alpha-helix structures and the NMR relaxation populations reflecting hydration water (T(2B) approximately 0-10 ms), myofibrillar water (T(21) approximately 35-50 ms), and also expelled "bulk" water (T(2) relaxation times >1000 ms). Accordingly, the present study demonstrates that definite structural changes in proteins during cooking of meat are associated with simultaneous alterations in the chemical-physical properties of the water within the meat.  相似文献   

3.
The effect of Gd-DTPA on the development in NMR relaxation of skeletal rabbit muscles post-mortem was investigated by dynamic low-field (0.47 T) relaxation measurements from 4 min post-mortem and until 23 h post-mortem. Twelve rabbits were included in the study, and half of the animals were administered 0.2 mmol of Gd-DTPA iv 15 min before sacrifice, while the other half was administered an isotonic salt solution. A significant effect of Gd-DTPA treatment corresponding to a 25% reduction in the T(1) relaxation time was observed. T(2) relaxation was decomposed into two components reflecting intra- and extracellular components (T(2)()alpha and T(2)()beta, respectively), and Gd-DTPA treatment was found to affect both components. However, around 150 min post-mortem a dramatic increase in the difference between control and Gd-DTPA-treated rabbits was observed in the relaxation time of the intracellular water population (T(2)()alpha). Electrical stimulation of the muscles resulted in a significantly earlier onset of the increased effect of Gd-DTPA on the T(2)()alpha population. The increased effect of Gd-DTPA treatment on the T(2)()alpha component is believed to reflect leakage of water from the muscle cells due to membrane destabilization, known to be promoted by electrical stimulation. Accordingly, the present study demonstrates how Gd-DTPA can be used for probing membrane integrity in post-mortem muscles known to be of importance for subsequent water distribution and final water-holding capacity.  相似文献   

4.
Origin of multiexponential T(2) relaxation in muscle myowater.   总被引：5，自引：0，他引：5  
To obtain a further understanding of the nature of the multiexponential T(2) relaxation seen in muscle tissue water (myowater), relaxation measurements were carried out on whole, minced, and homogenized pork of three different qualities with regard to water-holding capacity (normal, red soft exudative, and dark firm dry). Whole, minced, and homogenized pork all resulted in multiexponential T(2) relaxation (three components) independently of the quality, even though microscopic studies on homogenized meat revealed considerable disruption of the macroscopic structure. This states that the relaxation behavior in meat cannot be explained by intra-/extracellular compartmentalization of the water as suggested in earlier studies. Subsequent studies of T(2) relaxation in either whole meat, where the structure integrity was changed by the introduction of dimethyl sulfoxide (membrane disruption) or urea (protein denaturation), or minced meat with added NaCl (inter-/intraprotein interactions) lead to the suggestion that in whole meat (i) the fastest relaxation component reflects water tightly associated with macromolecules, (ii) the intermediate relaxation component reflects water located within highly organized protein structures, for example, water in tertiary and/or quaternary protein structures and spatials with high myofibrillar protein densities including actin and myosin filament structures, and (iii) the slowest relaxation component reflects the extra-myofibrillar water containing the sarcoplasmatic protein fraction. Finally, relaxation patterns in heat-set gels of superprecipitated actomyosin and bovine serum albumin similar to that identified in whole meat support the proposed nature of T(2) relaxation in muscle myowater.  相似文献   

5.
SDS-PAGE banding patterns of myofibrillar protein samples from turkey breast muscle with pH < or =5.8 at 15 min post-mortem (rapid glycolyzing) contained 133, 142, and 165 kDa bands that were absent in samples from carcasses with pH >6.0 at 15 min post-mortem (normal glycolyzing). These extra protein bands contained fragments of myosin as identified by Western blot analysis. Myosin fragments were also observed in protein samples from breast muscle not allowed to cool until 110 min post-mortem (delay chilled). In addition to myosin degradation, neublin degradation was more extensive in samples from rapid glycolyzing carcasses than for normal controls. Creatine kinase and glycogen phosphorylase were present in myofibrillar protein extracts of rapid glycolyzing carcasses in higher quantities than in normal controls. Results of this study provide insight into the molecular basis for previously reported reductions in meat quality of rapid glycolyzing and delay chilled turkey meat.  相似文献   

6.
Eighteen proteins and peptides that were found to change post-mortem in Longissimus dorsi from pig muscle were identified by the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The 18 peptides originate from 9 different proteins including the 3 structural proteins (actin, myosin heavy chain, and troponin T) and the 6 metabolic proteins glycogen phosphorylase, creatine kinase, phosphopyruvate hydratase, myokinase, pyruvate kinase, and dihydrolipoamide succinyltransferase. The molecular weight and estimated sequence length of the identified spots show that these fragments result from proteolytic activity in meat. Identification of the parent proteins and the enhanced post-mortem appearance of the degradation products make these specific peptides good candidates for meat quality markers, and further studies of these specific fragments will lead to a better understanding of the proteolytic activities involved in the post-mortem conversion of muscle to meat.  相似文献   

7.
A total of 120 4-week-old broiler chickens were allotted to 12 pens and fed one of three diets including control, oxidized diet (5% oxidized oil), or antioxidant-added diet (500 IU vitamin E) for 2 weeks. Blood samples were collected at the end of feeding trial, and breast muscles were sampled immediately after slaughter. Breast meats were also collected 24 h after slaughter and used for meat quality measurements. Oxidative stress in blood, lipid and protein oxidation, and sarcoplasmic reticulum Ca2(+)-ATPase (SERCA) activity of breast muscle were determined. The oxidized diet increased oxidative stress in blood and increased carbonyl content in breast meat compared with the other two dietary treatments (P < 0.05). Lipid oxidation of breast muscles with the antioxidant-supplemented diet was lower than that with the oxidized and control diet groups (P < 0.05). Meat from birds fed the oxidized diet showed higher drip loss after 1 and 3 days of storage and greater 0-1 h post-mortem pH decline (P < 0.05). Significant differences in specific SERCA activity in breast muscles from birds fed control and oxidized diets (P < 0.05) were detected. This suggested that dietary oxidized oil induced oxidative stress in live birds and increased lipid and protein oxidation in breast muscle. Decrease in SERCA activity in breast muscles due to oxidative stress in live animals accelerated post-mortem glycolysis, which sped the pH drop after slaughter and increased drip loss, indicating that oxidation of diet can cause PSE-like (pale, soft, and exudative) conditions in broiler breast muscles.  相似文献   

8.
Postmortem changes in rabbit muscle tissue with different glycogen status (normal vs low) were followed continuously from 13 min postmortem until 8 h postmortem and again 20 h postmortem using simultaneous magic angle spinning (1)H and (31)P NMR spectroscopy together with measurement of the transverse relaxation time, T(2), of the muscle water. The (1)H metabolite spectra were measured using the phase-altered spinning sidebands (PASS) technique at a spinning rate of 40 Hz. pH values calculated from the (31)P NMR spectra using the chemical shifts of the C-6 line of histidine in the (1)H spectra and the chemical shifts of inorganic phosphate in the (31)P spectra confirmed the different muscle glycogen status in the tissues. High-resolution (1)H spectra obtained from the PASS technique revealed the presence of a new resonance line at approximately 6.8 ppm during the postmortem period, which were absent in muscles with low muscle glycogen content. This new resonance line may originate from the aminoprotons in creatine, and its appearance may be a result of a pH effect on the exchange rate between the amino and the water protons and thereby the NMR visibility. Alternatively, the new resonance line may originate from the aromatic protons in tyrosine, and its appearance may be a result of a pH-induced protein unfolding exposing hydrophobic amino acid residues to the aqueous environment. Further studies are needed to evaluate these hypotheses. Finally, distributed analysis of the water T(2) relaxation data revealed three relaxation populations and an increase in the population believed to reflect extramyofibrillar water through the postmortem period. This increase was significantly reduced (p < 0.0001) in samples from animals with low muscle glycogen content, indicating that the pH is controlling the extent of postmortem expulsion of water from myofibrillar structures. The significance of the postmortem increase in the amount extramyofibrillar water on the water-holding capacity was verified by centrifugation, which showed a reduced centrifugation loss in muscles with low preslaughter glycogen status (0.9 vs 1.9%, p = 0.07).  相似文献   

9.
李乔  马纪兵  余群力  韩玲 《核农学报》2022,36(7):1413-1424
为研究宰后NO-AMPK通路对牛肉蛋白特性及肉品质的影响,以一氧化氮合成酶(NOS)抑制剂L-NAME、一磷酸腺苷活化蛋白激酶(AMPK)抑制剂Compound C处理的牛臀股四头肌为研究对象,生理盐水处理作为对照,将肉样与处理液在4℃条件下1:1(g:mL)混合浸泡12 h后,在成熟0、6、12、24、48、72、120 h测定肉样的蛋白功能特性、保水性、嫩度以及肌肉组织结构等相关指标。结果显示,L-NAME组中一氧化氮NO含量在6~72 h显著低于对照组(P<0.05);3组中AMPK活性先增大后减小,L-NAME组和Compound C组中AMPK活性在6 h分别比对照组降低14.78%和26.75%;L-NAME组中总蛋白溶解性在6、24 h低于Compound C组,高于对照组(P<0.05),表面疏水性在12、48 h显著低于对照组(P<0.05)并高于Compound C组;L-NAME组中剪切力仅在48 h显著高于对照组并低于Compound C组(P<0.05);对照组中肉样蒸煮损失最大,Compound C组中肉样蒸煮损失最小。以上结果表明,牛肉宰后成熟过程中,NO能够通过AMPK通路改善牛肉嫩度,但对其保水性产生了不利影响。本研究结果可为牛肉宰后能量代谢及肉品质控制相关研究提供一定的理论基础。  相似文献   

10.
Fourier transform infrared (FT-IR) microspectroscopy and low-field (LF) proton NMR transverse relaxation measurements were used to study the changes in protein secondary structure and water distribution as a consequence of aging (1 day and 14 days) followed by salting (3%, 6%, and 9% NaCl) and cooking (65 degrees C). An enhanced water uptake and increased proton NMR relaxation times after salting were observed in aged meat (14 days) compared with nonaged meat (1 day). FT-IR bands revealed that salting induced an increase in native beta-sheet structure while aging triggered an increase in native alpha-helical structure before cooking, which could explain the effects of aging and salting on water distribution and water uptake. Moreover, the decrease in T2 relaxation times and loss of water upon cooking were attributed to an increase in aggregated beta-sheet structures and a simultaneous decrease in native protein structures. Finally, aging increased the cooking loss and subsequently decreased the final yield, which corresponded to a further decrease in T2 relaxation times in aged meat upon cooking. However, salting weakened the effect of aging on the final yield, which is consistent with the increased T2 relaxation times upon salting for aged meat after cooking and the weaker effect of aging on protein secondary structural changes for samples treated with high salt concentration. The present study reveals that changes in water distribution during aging, salting, and cooking are not only due to the accepted causal connection, i.e., proteolytic degradation of myofibrillar structures, change in electrostatic repulsion, and dissolution and denaturation of proteins, but also dynamic changes in specific protein secondary structures.  相似文献   

11.
Proteome analysis was successfully applied to study the alterations in fish muscle proteins during ice storage. The processes occurring during post-mortem metabolism are known to lead to characteristic changes in the texture and taste of fish muscle. Endogenous proteases are anticipated to play the major role in these processes, although the exact mechanisms during fish meat tenderization have yet to be depicted. Protein changes in cod (Gadus morhua) muscle were followed during 8 days of storage. Within the partial proteome (pI 3.5-8.0, MW 13-35 kDa) significant changes were found in 11 protein spots. In nine protein spots the intensity increased, and for eight of these the increases were significant (p < 0.05) within the first 2 h post-mortem. In contrast, two protein spots decreasing in intensity showed significant (p < 0.03) changes after 8 days, thereby indicating that in the fish muscle different biochemical processes are involved in the protein changes observed post-mortem.  相似文献   

12.
The role of phospholipase A2 in the induction of drip loss from pig muscle has been investigated. In samples from porcine M. longissimus dorsi, total PLA2 activity as well as mRNA and protein levels of the group VIA iPLA2 (iPLA2-VIA) increased during the initial 4 h post-mortem period. Morphological studies of porcine muscle showed that at 4 h post-mortem, gaps had formed between muscle fibers and that the sarcolemma membrane borders appeared blurred. At the same time iPLA2-VIA protein levels were increased inside muscle fibers and at the sarcolemma. iPLA2-VIA mRNA abundance in samples from different breeds of pigs with variations in drip loss revealed no clear correlation between drip loss level and iPLA2-VIA expression. Together, these data indicate that during the post-mortem period, iPLA2-VIA expression and activity is increased at the muscle fiber membranes. PLA2 activity may affect membrane permeability and consequently the progression of drip formation in porcine muscle.  相似文献   

13.
Nuclear magnetic resonance (NMR) measurements were carried out on pork longissimus muscle, which pre rigor had been manipulated to various muscle lengths, to investigate the relationship between the microstructure of meat and the NMR T(2) relaxation. Distributed exponential analysis of the NMR T(2) relaxation data revealed the existence of three distinct water populations: T(2b), T(21), and T(22). A high, significant correlation was found between the T(21) time constant and the sarcomere length (r = 0.84) and calculated ration of myofilament lattice volume in the I-band and A-band regions, respectively (r = 0.84), considering sigmoid relationships. The result implies that the T(21) time constant mainly is determined by the structure of the myofilament lattice and so strongly supports a previously proposed theory that the T(21) population corresponds to water located within a highly organized myofibrillar protein matrix including actin and myosin filament structures. A high correlation was also found between the T(22) population and the water-holding capacity (WHC) (r = 0.76), which suggests that the WHC is mainly determined by the amount of loosely bound extramyofibrillar water. However, the correlation between NMR T(2) parameters and WHC was further increased (r = 0.84) by including the T(21) time constant in the correlation analysis. This implies that the formation of drip loss is an ongoing process involving the transfer of water from myofibrils to the extracellular space and is affected by structural features at several levels of organization within the muscle tissue. This study demonstrates the advantages of NMR T(2) relaxation as an effective technique for obtaining further understanding of the relationship between the microstructure of meat, its WHC, water mobility, and water distribution.  相似文献   

14.
为了探讨超声辅助变压滚揉对原料肉腌制过程蛋白质结构及水分含量的影响,该研究以鸡胸肉为原料分别进行静置腌制(对照)、真空滚揉(vacuum tumbling,VT)腌制和超声辅助变压滚揉(pressure-transform tumbling curing assisted by ultrasound,PTU)腌制处理100 min,采用拉曼光谱、低场核磁共振(low-field nuclear magnetic resonance,LF-NMR)、扫描电镜(scanning electron microscopy,SEM)和透射电镜(transmission electron microscope,TEM)测定不同腌制处理后鸡肉肌原纤维蛋白的二级结构、肌肉中水分含量以及肌肉组织微观结构的变化.结果显示,经过VT和PTU腌制处理的鸡肉其各项指标较对照组均发生显著变化(P＜0.05),其中拉曼光谱分析显示PTU处理的鸡肉肌原纤维蛋白质二级结构组成发生更显著变化,α-螺旋含量降低(P＜0.05),β-折叠和-转角的含量显著增加(P＜0.05);LF-NMR结果显示PTU处理的鸡肉的结合水横向弛豫时间和不易流动水横向弛豫时间较VT处理显著减小(P＜0.05),而对应的结合水峰面积百分比和不易流动水峰面积百分比都较其他处理组增加(P＜0.05).扫描电镜和透射电镜研究结果也显示经PTU处理的鸡肉组织结构破坏最严重,Z线以及肌原纤维膜降解最为明显.综上,PTU处理能够通过改变蛋白质结构、水分存在状态分布以及微观结构的变化来加速原料鸡肉腌制效率,改善嫩度和提高保水性;研究结果为PTU技术在肉制品加工中的应用及相关设备的开发提供理论依据.  相似文献   

15.
代谢组学在肉及肉制品品质监测中的应用   总被引：2，自引：1，他引：1  
代谢组学是通过研究机体受外界干扰前后小分子代谢物(分子量1 500 Da)的变化,进而探究其代谢机制的新兴科学。近年来,代谢组学在肉品科学研究领域受到广泛关注。但目前基于该技术监测宰前因素(遗传因素、肌肉部位及饲喂方式)及宰后成熟(时间、方式)对肉及肉制品品质影响的相关研究仍缺乏系统总结。同时,代谢组学技术的引入,也为肉品货架期预测、肉制品加工工艺优选、产地溯源及真伪鉴别提供了新的思路。因此,该研究概述了近年来代谢组学常用的分析检测技术(核磁共振技术、气相色谱质谱联用技术、液相色谱质谱联用技术)及数理统计方法(主成分分析、偏最小二乘判别分析等),重点对代谢组学在肉品生产诸多环节(动物饲喂、屠宰、加工等)中的最新研究进展进行综述,最后总结了目前肉品代谢组学研究中存在的代谢产物检测有限、试验重复性差等问题并认为多组学联合分析是监测肉品品质的未来发展方向,以期为其在肉品科学领域的应用提供参考。  相似文献   

16.
The objective of this study was to investigate the influence of heating rate on myowater dynamics and protein secondary structures in three pork qualities by proton NMR T2 relaxation and Fourier transform infrared (FT-IR) microspectroscopy measurements. Two oven temperatures at 100 degrees C and 200 degrees C corresponding to slow and fast heating rates were applied on three pork qualities (DFD, PSE, and normal) to an internal center temperature of 65 degrees C. The fast heating induced a higher cooking loss, particularly for PSE meat. The water proton T21 distribution representing water entrapped within the myofibrillar network was influenced by heating rate and meat quality. Fast heating broadened the T21 distribution and decreased the relaxation times of the T21 peak position for three meat qualities. The changes in T21 relaxation times in meat can be interpreted in terms of chemical and diffusive exchange. FT-IR showed that fast heating caused a higher gain of random structures and aggregated beta-sheets at the expense of native alpha-helixes, and these changes dominate the fast-heating-induced broadening of T21 distribution and reduction in T21 times. Furthermore, of the three meat qualities, PSE meat had the broadest T21 distribution and the lowest T21 times for both heating rates, reflecting that the protein aggregation of PSE caused by heating is more extensive than those of DFD and normal, which is consistent with the IR data. The present study demonstrated that the changes in T2 relaxation times of water protons affected by heating rate and raw meat quality are well related to the protein secondary structural changes as probed by FT-IR microspectroscopy.  相似文献   

17.
Low-field (LF) (1)H NMR T 2 relaxation measurements were used to study changes in water distribution in lean (Atlantic cod) and fatty (Atlantic salmon) fish during salting in 15% NaCl and 25% NaCl brines. The NMR data were treated by PCA, continuous distribution analysis, and biexponential fitting and compared with physicochemical data. Two main water pools were observed in unsalted fish, T 21, with relaxation times in the range 20-100 ms, and T 22, with relaxation times in the range 100-300 ms. Pronounced changes in T 2 relaxation data were observed during salting, revealing changes in the water properties. Salting in 15% brine lead to a shift toward longer relaxation times, reflecting increased water mobility, whereas, salting in saturated brines had the opposite effect. Water mobility changes were observed earlier in the salting process for cod compared to salmon. Good linear correlations ( F 相似文献   

18.
为阐明热风干过程中风干肉水分迁移机制,利用氢质子低场核磁共振弛豫研究热风干过程中(温度35℃、湿度60%、风速3 m·s-1)风干肉内部和外部中水分迁移及水-蛋白相互作用模式。结果表明,在风干过程中风干肉中水分的主要存在形式为不易流动水,其不断从肌原纤维网络内迁移到网络外,水分含量显著下降。风干肉外部中结合水、不易流动水和自由水的弛豫时间均显著下降(P < 0.05),表明风干肉外部中水-蛋白相互作用模式发生改变。而风干肉内部中结合水和自由水的弛豫时间无显著变化(P > 0.05),表明风干过程中风干肉内部和外部中水-蛋白相互作用不同,风干肉表皮的温度较内部温度高,且表皮水分含量下降速度较内部的水分含量下降速度快,导致了硬壳的形成。综上所述,风干过程中风干肉外部的温度高且水分脱除快,导致了风干肉内外部中水-蛋白相互作用不同。本研究结果为调控风干过程中水分迁移速度,实现风干肉工业化加工提供了理论依据。  相似文献   

19.
Solid-state (13)C cross-polarization (CP) magic-angle spinning (MAS) nuclear magnetic resonance (NMR) experiments are carried out for the first time on rapidly frozen muscle biopsies taken in M. longissimus in vivo and at 1 min, 45 min, and 24 h post-mortem from three pigs. Two of the pigs were CO(2)-stunned (control animals), and one was pre-slaughter-stressed (treadmill exercise) followed by electrical stunning to induce difference in metabolism post-mortem. (13)C resonance signals from saturated and unsaturated carbons in fatty acids, carboxylic carbons, and carbons in lactate and glycogen are identified in the solid-state NMR spectra. The (13)C CP MAS spectra obtained for post-mortem samples of the stressed, electrically stunned pig differ significantly from the post-mortem control samples, as the intensity of a resonance line appearing at 30 ppm, assigned to carbons of the methylene chains, is reduced for the stressed pig. This spectral difference is probably due to changes in lipid mobility and indicates altered membrane properties in the muscle of the stressed/electrically stunned animal when compared with the control animals already 1 min post-mortem. In addition, the post-mortem period changes in glycogen carbons can be estimated from the (13)C CP MAS spectra, yielding a correlation of r = 0.74 to subsequent biochemical determination of the glycogen content.  相似文献   

20.
The molecular motion of water was studied in glucono-δ-lactone-acidified skim milk powder (SMP) solutions with various pH values and dry matter contents. NMR relaxometry measurements revealed that lowering the pH in SMP solutions affected 17O and 1H T2 relaxation rates almost identically. Consequently, the present study indicates that the proteins present in the samples do not affect the 1H relaxation behavior markedly, even at relatively high SMP concentrations (15-25%). Comparison of rheological measurements and NMR measurements suggested that the collapse of κ-casein during acidification could contribute to the initial decrease in 17O and 1H relaxation rate in the pH range between 6.6 and 5.5 for 15% SMP and in the pH range between 6.6 and 5.9 for 25% SMP. However, below pH 5.5 the viscosity and 17O and 1H NMR relaxation rates did not correlate, revealing that the aggregation of casein micelles, which increases viscosity below pH 5.5, does not involve major repartitioning of water.  相似文献   

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