Contribution of postmortem muscle biochemistry to the delivery of consistent meat quality with particular focus on the calpain system

Tenderness has been repeatedly reported as the most important quality aspect of meat. However, a number of studies have shown that a significant portion of retail meat can be considered tough. As a consequence, a significant consumer segment is willing to pay a premium for guaranteed tender meat. However, apart from measuring the shear force, there is no reliable method to predict tenderness. Most of the branded meat programs therefore attempt to ensure eating quality by controlling some of the factors that affect tenderness. Meat tenderness is determined by the amount and solubility of connective tissue, sarcomere shortening during rigor development, and postmortem proteolysis of myofibrillar and myofibrillar-associated proteins. Given the effect of postmortem proteolysis on the muscle ultrastructure, titin and desmin are likely key substrates that determine meat tenderness. A large number of studies have shown that the calpain proteolytic system plays a central role in postmortem proteolysis and tenderization. In skeletal muscle, the calpain system consists of at least three proteases, μ-calpain, m-calpain and calpain 3, and an inhibitor of μ- and m-calpain, calpastatin. When activated by calcium, the calpains not only degrade subtrates, but also autolyze, leading to loss of activity. m-Calpain does not autolyze in postmortem muscle and is therefore not involved in postmortem tenderization. Results from a number of studies, including a study on calpain 3 knockout mice, have shown that calpain 3 is also not involved in postmortem proteolysis. However, a large number of studies, including a study on μ-calpain knockout mice, have shown that μ-calpain is largely, if not solely, responsible for postmortem tenderization. Research efforts in this area should, therefore, focus on elucidation of regulation of μ-calpain activity in postmortem muscle. Discovering the mechanisms of μ-calpain activity regulation and methods to promote μ-calpain activity should have a dramatic effect on the ability of researchers to develop reliable methods to predict meat tenderness and on the meat industry to produce a consistently tender product.     

细胞凋亡对肌肉宰后嫩化的贡献

嫩度是肉品最重要的品质指标,而成熟是改善肉的嫩度的重要途径。肉成熟嫩化的原因普遍认为是肌细胞的细胞骨架降解造成的。目前,细胞骨架降解的机理研究方面报道较多的是钙激活酶系统、组织蛋白酶系统和蛋白酶体,而有关细胞凋亡酶在肉成熟中的作用国内外还鲜见报道。本文综述了宰后肌细胞内生理条件的变化,以及细胞可能选择的死亡方式,即凋亡;概述了能作为caspase酶家族底物的几类主要的肌细胞骨架蛋白;对常用的几类诱导细胞凋亡的细胞毒素进行了介绍;最后,对细胞毒素注射应用于宰后牛肉品质形成机理研究的潜在意义进行了探讨。     

Factors influencing the variation in tenderness of seven major beef muscles from three Angus and Brahman breed crosses

Beef carcasses (n=30) from 3/4 Angus (A)×1/4 Brahman (B), 1/4A×3/4B, and 1/2A×1/2B F(1) crosses were used to evaluate breed type, electrical stimulation, and postmortem aging on the M. semimembranosus (SM), M. semitendinosus (ST), M. biceps femoris (BF), M. vastus lateralis (VL), M. gluteus medius (GM), M. longissimus dorsi lumborum (LD), and M. triceps brachii (TB). Shear force values decreased with increased postmortem aging to a greater extent in steaks from 3/4A×1/4B than steaks from the other breed types. Shear force values for steaks from the round (SM, ST, BF, VL) were higher than steaks from the loin (LD, GM) and chuck (TB) for both electrically stimulated and non-electrically stimulated muscles. In the LD muscle, calpastatin activities were similar among breed types. Muscle type played the greatest role in determining tenderness.     

Influence of ultimate pH on bovine meat tenderness during ageing

The aim of this work was to evaluate the influence of ultimate pH and ageing at 2±2°C on the tenderness of beef. The m.longissimus thoracis et lumborum from 23 young bulls excised at 28 h post mortem were grouped into: Normal (pH 5.5 to 5.8) moderate DFD (mod DFD) (5.8p<0.05) lower in the DFD group than in the Normal one and a significant (p<0.001) linear relationship was found between ultimate pH and tenderness as evaluated by both methods. The myofibrillar protein solubility (MPS) at pH 7.0 was significantly higher in the DFD group at all post mortem times whereas for myofibrillar fragmentation index (MFI) higher values (p<0.05) were only found in the DFD group at day 1. In all three groups toughness decreased (p<0.05) from 1 to 6 days, with no further decrease to 13 days. This decrease in toughness was associated with an increase of MFI in all pH groups. Collagen solubility in all three groups was not affected by ageing. No significant (p>0.05) differences in soluble collagen and myofibrillar protein solubility (MPS) at pH 5.5 were found between the pH groups at any time. The tenderness evaluated by both methods was significantly (p<0.05) related to MFI, MPS at pH 7.0, cooking loss and juiciness. Total and soluble collagen, sarcomere length, intramuscular fat and MPS at pH 5.5 were not significantly (p>0.05) related with tenderness.     

Sensory evaluations of porcine longissimus dorsi muscle: Relationships with postmortem meat quality traits and muscle fiber characteristics

The objective of this study was to investigate sensory evaluations and their relationships with meat quality measurements and histochemical characteristics in both fresh and cooked pork. Based on the results, postmortem meat quality traits were closely related to almost all the evaluated sensory attributes. With regard to histochemical characteristics, muscle fiber area was related to both fresh- (r = 0.18, P < 0.05) and cooked-meat color (r = −0.24, P < 0.01) as well as abnormal flavor intensity (r = 0.25, P < 0.01), and muscle fiber composition was associated with fresh pork color and taste acceptability after cooking. There were no significant relationships (P > 0.05) between type IIa muscle fiber content and the evaluated sensory attributes; however, good meat sensory quality was partially explained by the percentage of type I fiber.     

Difference in tenderness and pH decline between water buffalo meat and beef during postmortem aging

The objective of this research was to determine the difference in tenderness and some characteristics of water buffalo meat and beef during postmortem aging. Five female crossbred water-buffalo (Philippine Carabao×Bulgarian Murrah) and five female crossbred cattle (Brahman×Philippine Native), were finished on the same diet for 6 months and slaughtered at 30 months of age. The muscle pH was measured at 40min, 3h, 7h, 24h, and 48h postmortem. Longissimus thoracis (LT) and semimembranosus (SM) muscles were excised at 2d postmortem, and shear force was measured at 2, 4, 7, and 14d postmortem. Glycogen and lactate concentrations were determined from 0, 2, and 4d LT samples, and myosin heavy chain type of buffalo and cattle LT was determined by ELISA methods. Myofibrillar protein degradation was also observed by SDS-PAGE and Western blotting of fast-type troponin T. Results showed that the buffalo meat had significantly lower shear force values compared to beef for LT and SM muscles, which was supported by a difference in troponin T degradation. Postmortem pH decline of buffalo meat was significantly slower than that of beef, which was confirmed by lactic acid concentrations, but was not explained by glycogen content. In addition, there was no significant difference in the ratio of slow to fast type muscle fibers in buffalo and cattle, indicating that myosin heavy chain type was not responsible for the difference in pH decline and tenderness between the buffalo meat and beef. This study demonstrated that the tenderness of water buffalo meat was superior to that of Brahman beef, which may have been due to the difference in pH decline and the subsequent effect on muscle protease activity.     

Effect of dietary ractopamine on tenderness and postmortem protein degradation of pork muscle

Twenty-four finishing pigs with a mean starting weight of 82 kg were assigned to two dietary regimens: (1) a corn–soybean meal basal diet (control; n = 12), and (2) the basal diet supplemented with 20 ppm ractopamine HCl (RAC; n = 12). After 28–30 days on the feeding trial, pigs were slaughtered, and the growth and carcass characteristics were measured. Furthermore, the 3rd–13th rib section of longissimus muscle was excised at 48 h postmortem, sliced into 19-mm thick chops, vacuum packaged, stored at 2 °C, and subjected to Warner–Bratzler shear force (WBSF) and electrophoretic tests after 2, 4, 7, 10, 14, and 21 days (postmortem). RAC feeding increased (P < 0.01) pig carcass weight and percent lean, but it also increased the day-2 muscle WBSF by 20% (P < 0.01). The shear force difference between control and RAC pig muscles gradually decreased and vanished by day 10 (P > 0.05) when both muscle groups became more tender. The muscle from RAC-fed pigs exhibited a slower protein degradation rate than muscle from the control animals, notably for proteins in the 15–45 kDa range. The results suggested that the tenderness difference between ractopamine-treated and control pig muscles was related to the proteolysis rate, and could be diminished with adequate postmortem ageing.     

宰后肉成熟过程中的风味变化

介绍宰后肉在成熟过程中风味的变化,分别阐述肌糖原、蛋白质、脂类、核苷酸等几类化合物对肉风味的贡献.生肉很少有香味.并且只具有一种类似血腥的味道,但在宰后肉成熟的过程中所产生的丰富的具有味觉属性的化合物.以及香味的前提物质和风味增强剂对肉的感官性质产生巨大影响.     

Effects of temperature conditioning on postmortem changes in physico-chemical properties in Korean native cattle (Hanwoo)

The objective of this study was to examine the changes in physico-chemical properties that occur after adjusting postmortem chilling temperatures in Hanwoo beef. The right sides of beef carcasses were chilled for 4h at 2°C, 4h at 12°C and 16h at 2°C. The left sides were used as controls, chilled for 24h at 2°C. It took 8h for the control carcasses to cool down to <10°C and 10h 20min for the treatment. Adjusting the chilling temperature could be effective in lowering the postmortem pH and glycogen content. Treatment muscles at least 8h postmortem had longer sarcomeres than the control (P<0.05). The shear force in treatment carcasses at day 1 was similar to that of the control at day 6. Alternate chilling temperature had no detrimental effect on drip, cooking, total loss or color. Total numbers of aerobic plate counts were not significantly different between the control and treatment.     

Influence of blade tenderization,moisture enhancement and pancreatin enzyme treatment on the processing characteristics and tenderness of beef semitendinosus muscle

The combined effect of blade tenderization (BT), moisture enhancement and enzymatic tenderization on drip loss, cook loss, Warner–Bratzler shear force (WBSF) and sensory characteristics of beef semitendinosus (ST) steaks from cattle under 30 months of age was investigated.     

Revisiting the conversion of muscle into meat and the underlying mechanisms

The conversion of muscle into meat is a complex process in which all mechanisms responsible for the development of meat qualities are very likely interdependent. Colour and flavour are thus both dependent on oxidative mechanisms. Oxidation and proteolysis are probably two processes involved in the development of meat tenderness. This paper reviewed the consequences of programmed cell death or apoptosis on muscle cells structure and biochemistry and on meat qualities as well. We therefore look at different new hypothesis susceptible to highlight the meat science field and provide new supports for a more dynamic meat research. One of them which would have appeared evident for our purpose since a decade, deals with the fact that, after animal bleeding, muscle cells have no other alternative to only enter the programmed cell death procedure or apoptosis. If we introduce an early phase corresponding to apoptosis, taking place before the rigor onset and overlapping it, we will see that the known consequences of that process bring forward possible answers to still unexplained observations. After an overview of the actual state-of-the-art in meat science, we will introduce the programmed cell death and its underlying mechanisms. We then described the strong analogies between the known consequences of apoptosis and the postmortem changes affecting a set of different muscle characteristics.     

Influence of myosin heavy chain isoform expression and postmortem metabolism on the ATPase activity of muscle fibers

The objective of this study was to determine the effects of postmortem muscle pH and temperature declines on the actomyosin ATPase activity of muscle fibers expressing different MyHC isoforms. Using a quantitative histochemical procedure to determine ATPase activity, the maximum actomyosin ATPase activity was determined on individual fibers classified by MyHC expression. Samples were collected from the red (RST) and white (WST) semitendinosus muscles at 3 min and 24 h postmortem from electrically stimulated (ES) and control (NS) pork carcasses. In samples taken at 3 min postmortem, type I fibers had the lowest ATPase activity staining and type 2X and 2B had the highest activity staining, with type 2A fibers intermediate. Postmortem time and carcass treatment did not influence the ATPase activity staining of type I muscle fibers. ATPase activity staining of 2A fibers was lower (p<0.001) in 24 h samples than in 3 min samples from ES carcasses. In 3 min and NS-24 h samples, RST type 2A fibers had lower (p<0.05) activities than type 2A fibers from the WST. In type 2X fibers, ATPase activity staining decreased (p<0.01) from 3 min to 24 h postmortem in ES carcasses. This decrease was more severe in WST 2X fibers compared to RST 2X fibers. ATPase activity staining in type 2B fibers did not decrease from 3 min to 24 h postmortem in NS carcasses. In ES carcasses, activity staining of 2B fibers decreased (p<0.0001) with time postmortem. The results of the experiment indicate that fibers expressing fast MyHC isoforms have a higher ATPase activity early postmortem than slow muscle fibers but are more prone to inactivation by a rapid pH decline.     

Effects of blade tenderization,aging method and aging time on meat quality characteristics of Longissimus lumborum steaks from cull Holstein cows

The effects of blade tenderization (BT), two aging methods (dry (D) and wet (W)), and aging time (2 and 23 d) on tenderness, color, and sensory properties of Longissimus lumborum muscles from 12 cull Holstein cows were evaluated. Dry-aged loins had higher combined trim and aging losses than control (C) for both D- and W-aging, mostly because of excess trim losses. BT steaks had WBSF of 33.13 N while C steaks had WBSF of 41.46 N (P = 0.09). Aging decreased WBSF. Blade tenderized steaks had higher cook loss than C steaks. Aging, W-aging, and BT × W-aging improved myofibrillar (sensory) tenderness scores. Aging and/or BT improves sensory panel tenderness cull cow Longissimus lumborum steaks. Aging and blade tenderization combined can increase tenderness and value of Longissimus steaks from cull Holstein cows.     

Water distribution and mobility in meat during the conversion of muscle to meat and ageing and the impacts on fresh meat quality attributes--a review

This paper reviews current knowledge on the distribution and mobility of water in muscle (myowater) ante- and post mortem and factors affecting these in relation to fresh meat quality parameters; water-holding capacity (WHC), tenderness and juiciness. NMR transverse relaxometry (T(2)) using bench-top Low-Field Nuclear Magnetic Resonance (LF-NMR) has characterised myowater distribution and mobility as well as structural features in meat which directly affect WHC. The current literature demonstrates that WHC is correlated to the water located outside the myofibrillar network (extra-myofibrillar). This review identifies the critical stages which affect the translocation of water into the extra-myofibrillar space and thus the potential for decreased WHC during proteolysis (the conversion of muscle to meat). This review discusses how the intrinsic properties of the water held within the meat could contribute to juiciness and tenderness. Tenderness has been shown to correlate to T(2), however breed and species differences made it difficult to draw firm conclusions. Further understanding of the inherent water properties of fresh meat and the factors affecting water distribution and mobility using NMR technologies will increase the understanding of WHC and tenderisation of fresh meat.     

Effects of blade tenderization on some quality characteristics of the muscles of mature ewes

 The effects of blade tenderization (BT) on some of the quality characteristics of longissimus dorsi (LD) and semimembranosus (SM) muscles from the carcasses of mature ewes were evaluated in this research. Values describing panel tenderness (P<0.01) and residue after chewing (P<0.05) were significantly improved by BT. Also, the number of chews before swallowing was significantly reduced (P<0.01) in the tenderized samples. The LD muscle had a significantly better (P>0.01) panel tenderness score and number of chews value than the SM muscle. BT influenced significantly (P<0.05) the initial yield force values of the Instron Warner-Bratzler test, which were 5.42±0.40 kg and 6.45±0.42 kg for tenderized and non-tenderized samples, respectively. Peak force and work done values were also significantly (P<0.01) affected by BT. The LD muscle had a lower peak force value (5.88±0.39 kg vs 7.06±0.43 kg) than the SM muscle. However, other Instron Warner-Bratzler test parameters were not significantly (P>0.05) affected by muscle type. The chewiness, hardness, peak force 2 and work done 2 values determined by the Instron compression test were significantly affected by muscle type. Also, BT resulted in lower values for chewiness, hardness, peak force 1 and 2, and work done 1 and 2 values, but the differences between tenderized and non-tenderized samples were not statistically significant (P>0.05). Received: 16 July 1998 / Revised version: 14 October 1998     

Effect of South African beef production systems on post-mortem muscle energy status and meat quality

Post-slaughter muscle energy metabolism meat colour of South African production systems were compared; steers (n = 182) of Nguni, Simmental Brahman crossbreds were reared on pasture until A-, AB-, or B-age, in feedlot until A–AB-age. After exsanguination carcasses were electrically stimulated (400 V for 15 s). M. longissimus dorsi muscle energy samples were taken at 1, 2, 4 and 20 h. Post-mortem samples for meat quality studies were taken at 1, 7 and 14 days post-mortem. Production systems affected muscle glycogen, glucose, glucose-6-P, lactic acid, ATP, creatine-P glycolytic potential (P < 0.05), with the muscles of feedlot carcasses having a faster glycolysis rate than pasture carcasses. Energy metabolites correlated (0.4 < r < 0.9) with meat colour (CIE, L*a*b*), (0.3 < r > 0.5) water holding capacity, drip loss, and Warner Bratzler shear force. Muscle energy only affected muscle contraction of the A-age-pasture system (shortest sarcomere length of 1.66 μm vs 1.75 μm highest WBS of 6 kg vs 5 kg 7 days post-mortem).     

Alternating frequency to increase the response to stimulation from medium voltage electrical stimulation and the effect on objective meat quality

The use of alternating frequencies during stimulation can increase stimulation response of a medium voltage electrical stimulation unit (MVS) by increasing the rate of pH decline. Various combinations of frequency modulation were tested in experiment 1 to determine the treatment resulting in the greatest stimulation response; the lowest initial pH, fastest rate of pH decline, highest temperature at pH 6 and the highest number of carcasses with a pH of 6 by 25(o)C and the treatment achieving the highest number of carcasses in the pH temp window (temperature at pH 6 between 18-25(o)C). The objective meat quality of these treatments compared to an unstimulated treatment was tested in experiment 2. Modulating the frequency (Hz) across the 6 segmented electrodes of the MVS by 10, 15, 25, 10, 15, 25Hz (Treatment 6, using a pulse width: 2.5ms, current: 1A) resulted in the greatest stimulation response. This treatment may be suitable for abattoirs that hot bone sheepmeat and require fast pH declines to ensure minimal cold shortening of meat. However, this treatment did not result in the tenderer meat despite the higher stimulation response. This treatment may have induced a greater number of contractions overall and therefore a greater pH decline response but resulted in less myofibrillar disruption compared to the other treatments due to a concomitant decreased force of contraction thus reducing potential tenderisation. Maintaining a constant frequency of 15Hz (Treatment 1; pulse width: 2.5ms, current: 1A) resulted in a higher number of carcasses in the pH temp window required (temperature at pH 6 between 18-25(o)C) in part A (P<0.05) and in addition to the higher tenderness levels this treatment may be more appropriate to satisfy the overall demands of abattoirs using these systems. This paper has also demonstrated electrical stimulation results in tenderer meat compared to unstimulated meat even after 30d of ageing (2.53±0.4 compared to 2.85±0.1 for the loin (M. longissimus thoracis et lumborum) (P<0.01) possibly due to a protective benefit of stimulation on meat tenderness. Overall, no detrimental effects of modulating frequency were observed on drip loss or retail colour display despite a greater rate of colour change observed with the modulated frequency treatment and the longer aged product.     

Genetic and environmental effects on the muscle structure response post-mortem

This paper reviewed the mechanisms by which glycolytic rate and pre-rigor stretching of muscle impact on meat quality. If muscle is free to shorten during the rigor process extremes in glycolytic rate can impact negatively on meat quality by inducing either cold or rigor shortening. Factors that contribute to variation in glycolytic rate include the glycogen concentration at slaughter and fibre type of the muscle. Glycolysis is highly sensitive to temperature, which is an important factor in heavy grain fed carcasses. An alternative solution to controlling glycolysis is to stretch the muscle pre-rigor so that it cannot shorten, thus providing an insurance against extremes in processing conditions. Results are presented which show a large reduction in variance (both additive and phenotypic) in tenderness caused by pre-rigor stretching. Whilst this did not impact on the heritability of shear force, it did reduce genotype differences. The implications of these results on the magnitude of genotype effects on tenderness is discussed.     

Purification and partial characterization of ostrich skeletal muscle cathepsin D and its activity during meat maturation

Cathepsin D was purified from ostrich (Struthio camelus) skeletal muscle using pepstatin-A chromatography. The enzyme was comprised of two subunits (29.1 and 14 kDa). The N-terminal amino acid sequence of both subunits were determined and showed high amino acid sequence identity to other cathepsin D homologs. Ostrich cathepsin D was optimally active at pH 4 and at a temperature of 45°C, and was strongly inhibited by pepstatin-A (K(i)=3.07×10(-9)M) and dithiothreitol. Cathepsin D activities from five ostriches were monitored over a 30-day period. Cathepsin D remained substantially active throughout the 30-day storage period with an average remaining activity of 112±8.57% at day 30 (mean value from 5 ostriches).     

生姜汁在鹅腿肉嫩化中的应用研究

以剪切力为指标研究生姜汁对鹅腿肉的嫩化效果,通过单因素实验对生姜汁浓度、pH、处理温度进行了研究,并且进一步通过正交实验选择出最佳嫩化工艺。结果表明:生姜汁对鹅腿肉的嫩化效果十分显著(P<0.01),生姜汁对鹅腿肉嫩化的最佳工艺条件是:生姜汁浓度6%、pH6.0、处理温度为50℃。