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1.
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   

2.
To investigate changes in myosin light chains (MyLCs) during postmortem aging of the bovine longissimus muscle, we performed two-dimensional gel electrophoresis followed by identification with matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The results of fluorescent differential gel electrophoresis showed that two spots of the myosin regulatory light chain (MyLC2) at pI values of 4.6 and 4.7 shifted toward those at pI values of 4.5 and 4.6, respectively, by 24 h postmortem when rigor mortis was completed. Meanwhile, the MyLC1 and MyLC3 spots did not change during the 14 days postmortem. Phosphoprotein-specific staining of the gels demonstrated that the MyLC2 proteins at pI values of 4.5 and 4.6 were phosphorylated. Furthermore, possible N-terminal region peptides containing one and two phosphoserine residues were detected in each mass spectrum of the MyLC2 spots at pI values of 4.5 and 4.6, respectively. These results demonstrated that MyLC2 became doubly phosphorylated during rigor formation of the bovine longissimus, suggesting involvement of the MyLC2 phosphorylation in the progress of beef rigor mortis. Keywords: Bovine; myosin regulatory light chain (RLC, MyLC2); phosphorylation; rigor mortis; skeletal muscle.  相似文献   

3.
One of the main biochemical changes that take place during the processing of dry-cured ham is the degradation of the muscle protein fraction, mainly due to the action of muscle enzymes. In the present study, the isolation and tentative identification of 137 fragments from myosin light chain 1 (MLC 1), together with 88 fragments originated from myosin light chain 2 (MLC 2), have been achieved for the first time in Spanish dry-cured ham, proving the intense proteolysis experienced by myofibrillar proteins after dry-cured processing. This study was carried out by use of proteomic technology for peptide identification, and the possible enzymes contributing to the degradation of these proteins were also further discussed.  相似文献   

4.
The thermal behaviors of myosin from bovine vastus intermedius (VI, predominantly red muscle) and semimembranosus (SM, predominantly white muscle) at pH 6.05 (ultimate pH of VI muscle) and 5.50 (ultimate pH of SM muscle) were compared. Differential scanning microcalorimetry and turbidity measurements were used to monitor changes in myosin during heating from 25 to 80 degrees C at 1 degrees C/min. VI and SM myosin heavy chain isoforms were identified on gradient SDS-PAGE. Endotherms of VI myosin at pH 6.05 had three transition temperatures (T(m)) of 45, 53, and 57 degrees C, whereas at pH 5.50 two transitions were observed at 42 and 59 degrees C. SM myosin had two T(m) values of 46 and 58 degrees C at pH 6.05 and T(m) values of 43 and 62 degrees C at pH 5.5. SM myosin at its ultimate pH was less heat stable than VI myosin at its ultimate pH; however, when SM and VI myosin were compared at the same pH, VI myosin was less stable.  相似文献   

5.
Identification of factors that determine meat tenderness is of high priority. The aim of this work was to develop a method that can detect indicators of proteolysis in meat early postmortem. The method was validated on pork samples. A procedure to detect differences of extractable lower molecular weight compounds after a prerigor freeze/thaw cycle of meat was developed using capillary electrophoresis. The procedure was able to separate 39 peaks in the electropherograms. Eight of the peaks were correlated (P < 0.1) to Warner-Bratzler shear forces 1 day postmortem (WB1). A multiple linear regression model explained 69% of the variation in WB1 using the areas of four peaks. Several of the peaks used in modeling WB1 were related to the at-slaughter activity of the calpain system. The results presented show that the developed method is able to detect indicators of proteolysis and tenderness at an early time point after slaughter. The method is a new tool intended for studies regarding the mechanisms of postmortem proteolysis and tenderization.  相似文献   

6.
Two-dimensional electrophoresis was used to investigate sarcoplasmic protein expression in pig Semimembranosus muscles sampled 20 min after slaughter. Two groups (light and dark) of 12 animals were selected from 1000 pigs, based on meat L values measured 36 h postmortem. Twenty-two proteins or fragments (p < 0.05) were differentially expressed. Muscles leading to darker meat had a more oxidative metabolism, indicated by more abundant mitochondrial enzymes of the respiratory chain, hemoglobin, and chaperone or regulator proteins (HSP27, alphaB-crystallin, and glucose-regulated protein 58 kDa). Conversely, enzymes of glycolysis were overexpressed in the lighter group. Such samples were also characterized by higher levels of glutathione S-transferase omega, which can activate the RyR calcium channels, and higher levels of cyclophilin D. This protein pattern is likely to have severe implications on postmortem metabolism, namely, acceleration of ATP depletion and pH fall and subsequent enhanced protein denaturation, well-known to induce discoloration.  相似文献   

7.
Conformational and structural changes of cod myosin at pH 2.5 and 11 and after subsequent pH readjustment to pH 7.5 were studied. Results suggest that on acid unfolding, the myosin rod may fully dissociate due to electrostatic repulsion within the coiled coil, while it does not dissociate at alkaline pH. Both pHs led to significant conformational changes in the globular head fraction of the myosin heavy chains, suggesting that it takes on a molten globular configuration. A large part of the myosin light chains are lost on both pH treatments. On pH readjustment to neutrality, the heavy chains take on a structural form similar to the native state with the coiled-coil rod reassociating from acid pH while leaving the globular head less packed, more hydrophobic and structurally less stable. The irreversible change brought about in the globular head region leads to the failure of light chains to reassemble onto it, a drastic loss in ATPase activity, and more exposure of reactive thiol groups. The acid and alkali processes therefore lead to substantial changes in the globular part of the myosin molecule and perhaps more importantly to different molecular changes in myosin, depending on which pH treatment is employed.  相似文献   

8.
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9.
超高压处理对绵羊肉嫩化机理的研究   总被引：14，自引：2，他引：14  
实验研究了超高压处理条件下绵羊肌肉感官特性、显微结构、钙激活酶(Calpains)粗酶活性和剪切力值的变化，并探讨了超高压处理对绵羊肌肉的嫩化机理。超高压处理后绵羊肌肉的感官特性发生变化，随处理压力升高，绵羊肌肉颜色变淡，出现轻微的类似蒸煮的成熟风味。在压力为400 MPa，保压时间为10 min的处理条件下，绵羊肌肉显微组织结构变化明显：肌节收缩，肌原纤维的Z线断裂，M线降解，I带变白。当压力水平在100～400 MPa范围变动时，随压力升高，Calpains粗酶活性显著下降(P＜0.01)；当压力达到400 MPa时，Calpains粗酶活性几乎失活。超高压处理后绵羊肉的剪切力值显著下降(P＜0.05)。实验结果表明，超高压处理促进了绵羊肉的嫩化。  相似文献   

10.
为检测和分析0.5和3.0Gy 2种剂量重离子辐照绵羊精子差异表达的蛋白质,用二维凝胶电泳分离精子蛋白,凝胶用硝酸银染色,PDQuest8.0检测分析差异表达蛋白斑点并经高效液相色谱串联离子阱质谱鉴定,结果显示:2种剂量凝胶图谱分析后得到8个共同差异蛋白斑点,鉴定为3种表达上调的蛋白质(谷氧还蛋白1、转录因子AP-2α...  相似文献   

11.
为探明高压和氯化钙（CaCl2）注射结合处理对僵直后期牛肉嫩度的影响,首先分别对CaCl2浓度、高压强度及保压时间对宰后36?h的牛背最长肌嫩度的影响进行分析,在3个单因素试验的基础上,采用Box-Behnken响应曲面中心组合设计法,对高压和CaCl2结合处理嫩化牛肉的工艺参数进行优化,并通过透射电镜对肌纤维的超微结构进行分析。结果表明：以CaCl2浓度、高压强度和保压时间为自变量,剪切力值为响应值,得到的二次多项式回归模型拟合度高（决定系数R2=0.9742）;高压强度、保压时间、CaCl2浓度、高压强度和保压时间的交互作用对牛肉嫩化效果极显著（P＜0.01）,高压强度和CaCl2浓度的交互作用对牛肉嫩化效果显著（P＜0.05）。高压和CaCl2结合处理嫩化牛肉最佳工艺为：高压强度241?MPa,CaCl2浓度0.24?mol/L（样品质量5%的注射量）,保压时间14?min。应用此工艺嫩化牛肉,和对照相比牛背最长肌剪切力值下降了52.98%,肌纤维间隙增大,肌节完整性遭受破坏,牛肉嫩度明显改善。  相似文献   

12.
Two-dimensional electrophoresis was used to compare Longissimus sarcoplasmic protein abundance between two groups (tough meat and tender meat), defined on the basis of extreme Warner-Bratzler shear force values measured on cooked pork. Fourteen protein spots differed in quantity (P<0.05) between the two groups and were identified. Adypocyte fatty acid binding protein and acyl-CoA binding protein involved in lipid traffic and in the control of gene expression regulating cell proliferation and differentiation, and Enoyl-CoA hydratase, aldose reductase and triosephosphate isomerase indirectly related to lipid metabolism were overrepresented in the tender group. The tender group was further characterized by increased levels of proteins involved in protein folding and polymerization (initiation factor elf-3beta, chaperonin subunit 2, profilin II). The results suggest that the lower post-cooking shear force could at least in part be related to muscle adipogenetic and/or myogenetic status of which the possible underlying mechanisms are discussed.  相似文献   

13.
适宜宰后成熟时间提高牦牛肉品质   总被引：1，自引：1，他引：0  
为了研究宰后成熟时间对牦牛肉品质的影响,该研究采集牦牛背最长肌,真空包装后进行成熟排酸,成熟期间取样测定pH值、剪切力、持水能力、色度,并通过标准化和动力学模型分析研究其变化规律。结果发现,除了黄色度b*以外,其余品质指标均在21d的成熟过程中有显著变化(P0.05)。相比于7 d成熟,延长至21 d的长时间成熟可将牦牛肉剪切力显著地降低47%(P0.05),并将蒸煮损失和红色度a*分别显著地提高28%和32%(P0.05)。通过对比线性函数、指数函数、二次函数对牦牛肉品质变化的预测效果,结果发现二次函数预测模型与牦牛肉宰后品质变化具有相对最好的拟合度(决定系数R2范围为0.90~0.98),其中pH值、剪切力、加压损失、亮度L*值随时间的变化函数呈现出凸函数特征,而蒸煮损失和a*值则呈现出凹函数特征。结果表明,相比于牛肉品质宰后预测中常用的线性函数和指数函数而言,二次函数更适用于牦牛肉品质变化的预测,而将牦牛肉宰后成熟时间延长至21 d可有效降低牦牛肉剪切力。研究可为牦牛肉品质控制提供参考。  相似文献   

14.
Chickens from a randomly bred genetic line were segregated into high and low growth rates and high and low water-holding capacities (WHCs). The objective of this study was to identify protein markers associated with slow and fast growth rates and low and high WHCs from water-soluble protein (WSP) and crude myofibrillar protein (CMP) extracts of chicken breast muscle. Proteins were fractionated using two-dimensional electrophoresis, and a total of 22 protein spots were selected, excised, and analyzed by in-gel tryptic digestion and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Proteins expressed in extracts from slow and fast growth rates and low and high WHCs included metabolic enzymes, such as creatine kinase, pyruvate kinase, triosephosphate isomerase, and ubiqitin; housekeeping proteins, such as heat shock protein; contractile proteins, such as myosin heavy chain; actin; and also MHC isoforms and actin isoforms. The mass spectra of 20 protein spots significantly matched (protein score >83; P < 0.05) an online database. In CMP, there were unique proteins that were present only in the fast-growth population: gi|118099530 , gi|20664362 , gi|71895043 , gi|114794125 , gi|297343122 , and gi|71895043 . This information identified protein markers associated with growth rate and water holding capacity. Some of those protein markers could be added to the chicken database.  相似文献   

15.
Addition of papain decreased the onset temperature and the rate at which G' developed during heat-induced gelation of arrowtooth flounder myosin. Frequency sweep results revealed that G' markedly decreased in proportion to the amount of papain added. However, use of E-64, a cysteine proteinase inhibitor, reversed the effects of papain and protected myosin heavy chain from degradation. DSC thermograms indicated papain significantly decreased the enthalpy required to induce myosin denaturation without significant changes in onset and maximum transition temperatures. Thermal denaturation kinetics indicated decreases in both the activation energy and the rate of myosin denaturation. CD studies revealed a rapid decrease in alpha-helical content, indicating the initial degradation of myosin molecules mostly occurred in the tail region. These results suggested that proteolysis affected thermal properties and reactivity of myosin during heating. Although myosin gel could be formed, structural disruption resulted in lower gelling ability and rigidity of the formed gel.  相似文献   

16.
Natural actomyosin (NAM) extracted in 0.6 M NaCl from hake fillets stored at -20 and -30 degrees C for up to 49 weeks was studied. The extracted protein decreased as storage progressed and became poorer in myosin while the proportion of actin remained constant. Two major peaks composed of myosin plus actin and actin plus tropomyosin plus troponins were obtained by size exclusion chromatography. SDS-PAGE analysis of the protein retained in the precolumn filter showed that there was protein aggregated by covalent bonding. Surface hydrophobicity increased while Ca(2+)-ATPase activity, apparent viscosity, and SH groups decreased as storage progressed. The loss of Ca(2+)-ATPase activity was due mainly to denaturation of the extracted myosin, whereas the minimum viscosity values occurred earlier and were not directly due to the lower proportion of myosin in the extracts. Thus, the extracted NAM exhibited changes during frozen storage. The temperature-dependent difference was mainly quantitative due to a smaller amount of protein extracted at -20 degrees C.  相似文献   

17.
本实验选择具有优良肉质的本地金华猪与肉质较差的引进品种皮特兰猪杂交所产的金皮F2代为研究对象,检测金皮F2代猪CAST（calpastatin, CAST）基因、MyoG(Myogenin, MyoG)基因的PCR-MspI多态性,并结合肌球蛋白重链（MHC）免疫组织化学方法,研究猪背腰最长肌的肌肉组织学特性,并对其进行图像分析。结果发现,金皮F2代杂交猪背腰最长肌MHCs阳性（慢肌）纤维的比例为7.62%, 快肌纤维为92.38%。CAST基因的PCR扩增产物为1404bp的片段,酶切后分为A(628bp+499bp+277bp),B（499bp+371bp+277bp+257bp）两种等位基因;MyoG基因的PCR扩增产物长度为1050bp,酶切后分为A（462 bp +408 bp +180 bp）和B（408bp+290bp+180bp+162bp）两种等位基因。.AA、AB和BB基因型频率分别为 0.1795、0.5897和0.2308,A和B的基因型频率分别为 0.4743和0.5256。CC、CD和DD的基因型频率为0.3671,0.5696和0.0633, C和D的基因型频率分别为0.6519和0.3481。统计结果表明,AA、AB、AB、CC、CD、DD基因对眼肌面积,系水率、pH值、导电率等屠宰性状均无显著的影响。CAST基因的不同基因型对肌纤维的轴比有显著影响（P<0.05）,AA单型有产生较多轴比大（近似椭圆形）肌纤维的倾向,而BB则控制形成更多的轴比小（近似于圆形）的肌纤维。具有AA单型的个体具有更多的肌间结缔组织,而具有BB单型的个体的肌间结缔组织较少（P<0.05）,AB单型的个介于两者之间。MyoG基因MspI酶切后的各种基因型对肌肉的肌纤维大小和密度具有显著影响,具有DD基因型的个体的肌纤维面积较大,而具有CC基因型者肌纤维密度最大。  相似文献   

18.
The effects of Trichoderma reesei tyrosinase-catalyzed cross-linking of isolated chicken breast myofibril proteins as a simplified model system were studied with special emphasis on the thermal stability and gel formation of myofibrillar proteins. In addition, tyrosinase-catalyzed cross-linking was utilized to modify the firmness, water-holding capacity (WHC), and microstructure of cooked chicken breast meat homogenate gels. According to SDS-PAGE, the myosin heavy chain (MHC) and troponin T were the most sensitive proteins to the action of tyrosinase, whereas actin was not affected to the same extent. Calorimetric enthalpy (DeltaH) of the major thermal transition associated with myosin denaturation was reduced and with actin denaturation increased in the presence of tyrosinase. Low-amplitude viscoelastic measurements at constant temperatures of 25 degrees C and 40 degrees C showed that tyrosinase substantially increased the storage modulus (G') of the 4% myofibrillar protein suspension in the 0.35 M NaCl concentration. The effect was the most pronounced with high-enzyme dosages and at 40 degrees C. Without tyrosinase, the G' increase was low. Tyrosinase increased the firmness of the cooked phosphate-free and low-meat chicken breast meat homogenate gels compared to the corresponding controls. Tyrosinase maintained gel firmness at the control level of the low-salt homogenate gel and weakened it when both salt and phosphate levels were low. Tyrosinase improved the WHC of the low-meat and low-salt homogenate gels and maintained it at the level of the corresponding controls of phosphate-free and low-salt/low-phosphate homogenate gels. Microstructural characterization showed that a collagen network was formed in the presence of tyrosinase. Keywords: Chicken myofibrillar proteins; protein modification; cross-linking; tyrosinase; gelation; thermal stability; texture; water-holding capacity; microstructure.  相似文献   

19.
The effect of laccase and transglutaminase (TG) on cross-linking, gelation, and thermal stability of salt-soluble chicken-breast myofibril proteins was investigated at pH 6. Both enzymes modified the protein pattern detected by SDS-PAGE. Identification of proteins by peptide mass mapping showed that myosin heavy chain (MHC) and troponin T were the most affected proteins. These proteins faded or disappeared as a function of the incubation time with both enzymes on SDS-PAGE. The molecular weight of actin was not, however, affected by either enzyme. The effects that the enzymes had on the gel formation of chicken-breast myofibrils were studied in 0.35 and 0.60 M NaCl solutions at 3% protein content and a constant temperature of 40 degrees C by using a small deformation viscoelastic measurement. TG substantially increased the storage modulus (G') of 3% protein in 0.35 M NaCl. Without the enzymes, gelation was insignificant in 0.35 M NaCl. The increased solubility of the proteins at 0.60 M NaCl intensified gelation with TG. G' increased 32 and 64% at dosages of 10 and 100 nkat of TG, respectively. Also, laccase increased G' of the gel in 0.60 M salt concentration. However, a high laccase dosage decreased the magnitude of G' below the control level. Differential scanning calorimetric (DSC) measurements indicated slightly reduced myosin heat stability after TG pretreatment and increased actin heat stability with both enzymes. Maximum transition temperatures did not alter with either enzyme.  相似文献   

20.
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   

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