Effect of post-mortem temperature on beef tenderness

Beef adductor muscles were incubated for 4 h post mortem at 10°C and for 4 h and 6 h post mortem at 30°C, 37°C and 42°C. Half of the muscles were cooked just after incubation and the other half was first stored for two days at 4°C and then cooked. Meat kept for 4 h or 6 h at 42°C and for 6 h at 37°C and cooked at once had a significantly (p<0·05) lower shear force than meat kept for 4 h at 37°C, 4 h at 30°C, 6 h at 30°C or 4 h at 10°C. The respective significant differences were also found when the meat was cooked two days after incubation. Organoleptic evaluation showed that meat incubated for 6 h at 37°C or for 4 h at 42°C was not significantly more tender than other samples. However, meat kept for 6 h at 42°C was more tender (p<0·5) than the other samples. After two days of storage, meat incubated for 6 h at 37°C and for 6 h at 42° was more tender (p<0·05) than meat kept for 6 h at 30°C. It was concluded that high temperature conditioning at 37°C or higher for 6 h (4 h at 42°C) just after slaughter makes meat more tender than conventional cooling systems.     

Titin content of beef in relation to tenderness

Steaks obtained from the longissimus dorsi muscle of 24 crossbred steers were subjected to four treatments (unaged raw, aged raw, unaged cooked, aged cooked) and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Titin migrated primarily as a single protein band in unaged raw samples (48 h post mortem), as a doublet in aged (16 days) raw samples, and as a triplet in unaged and aged cooked samples. Total titin band density remained constant among steaks that varied widely in Warner-Bratzler shear value, suggesting that beef steaks varying in tenderness contain the same amount of titin. It is concluded that titin content, as determined by gel electrophoresis, does not distinguish 'tough' from 'tender' beef.     

Effects of early post-mortem pH and temperature on beef tenderness

The tenderness of loin steaks, prepared from beef sides after chilling, is strongly influenced by muscle temperature in the first 3 h after slaughter. Maintenance of about 37°C within the Longissimus muscle during this time, whether by heavy fat cover or by ambient-temperature manipulation, results in appreciable tenderness enhancement. Early post-mortem muscle pH (which varies over a wide range) also affects tenderness significantly; provided early and exceptionally fast chilling does not induce cold shortening, slow glycolysis promotes tenderness. Low-frequency (2 Hz) electrical stimulation, which accelerates glycolysis but causes negligible tissue disruption, significantly toughens the loin; in its normal mode (50–60 Hz), therefore, stimulation produces its desirable tenderising effect mainly—and perhaps solely—by fibre fracture.     

钙蛋白酶系统对肉的嫩化作用研究进展

肉的嫩度是肉品质的一个重要指标,严重影响肉的食用价值和经济价值.钙蛋白酶系统通过水解肌原纤维蛋白,在宰后肌肉嫩化中起着决定性的作用.综述钙蛋白酶系统(包含钙蛋白酶、钙蛋白酶抑制蛋白和钙蛋白酶激活蛋白3部分)对肉嫩化的作用机制和研究现状,并探讨研究前景.     

Tensile properties of cooked beef in relation to rigor temperature and tenderness

A study of the tensile properties of cooked strips from beef sternomandibularis which had gone into rigor at various temperatures showed no clear relationships with earlier observed variations in shear force with rigor temperature. A consistent feature of the load extension curves was a sudden increase in extensibility at a ‘change point’ of about 1 kg/cm². Some curves, particularly those for strips cooked under restraint, showed a third intermediate extensibility. Evidence points to modification of a myofibrillar component as the reason for the sudden changes.     

Calcium chloride marination effects on beef steak tenderness and calpain proteolytic activity

A study was conducted in three phases to examine the effect of calcium chloride marination on tenderness. Steaks obtained 5 days postmortem were marinated in a 150 mM calcium chloride solution for 24 h and 48 h in phase 1, and for 48 h in phases 2 and 3. The steaks utilized were obtained from mature cows 8-11 years of age-phase 1; four control and four β-agonist fed steers-phase 2; and three Peidmontese and two Nelore 18-month-old steers-phase 3. Data were analyzed by analysis of variance for a split-plot design. In phase 1, marination failed to improve (P > 0·05) shear force values. However, shear force values were less than 5 kg which was uncommonly low for mature cows. In phase 2, marination improved (P < 0·05) meat tenderness regardless of diet. Yet, the steaks from the β-agonist fed steers remained less tender, even after marination, than the steaks from the control steers. In phase 3, shear force requirements were decreased (P < 0·01) with marination. Also, the activities of m-calpain and calpastatin decreased (P < 0·05) with calcium marination. It appeared that the improvement in tenderness was through the activation of m-calpain.     

Effect of ion fluid injection on beef tenderness in association with calpain activity

Sodium pyrophosphate plus sodium chloride (PPi) was injected into pre-rigor, hot boned biceps femoris (BF) and semimembranosus (SM) muscle from 12 heifer carcasses. The PPi injection caused higher pH values between 10 and 48 h post-mortem than found in the controls for both muscles (P<0.05). PPi injection resulted in faster decreases in the activities of μ-calpain and calpastatin than in the control for both muscles with time post-mortem (P<0.05). There were significant differences between treatments in both the BF and SM (P<0.05). There was evidence that PPi-injection elevated pH, and accelerated activation of calpains, resulting in improved tenderness. The rates of degradation of titin and troponin-T as well as the appearance of 95 and 30 kDa peptides were faster in the PPi-injected muscles than the controls. PPi-injection elevated muscle pH, which was attributed to acceleration of the calpain activation. It is concluded that PPi-injection improved beef tenderness by accelerating activation of calpain.     

Effect of stress-induced high post-mortem pH on protease activity and tenderness of beef

Forty-four Swiss Brown young bulls were stressed by regrouping unfamiliar animals before slaughter. M. longissimus thoracis (6-9th ribs) of carcasses were analysed for post-mortem pH, protease activities (m- and α-calpain, calpastatin and cathepsin B + L), Warner-Bratzler shear force and sensory tenderness and juiciness. Muscles were classified into three groups, according to ultimate pH values: > 6.3, 6.3-5.8 and < 5.8. The most significant difference related to high pH was a higher activity of m-calpain at 7th day post mortem. It was also found that meat showing the highest pH was significantly more tender and juicy. Sensory tenderness was highly correlated with activity of m-calpain at 7th day post mortem (r = 0.776) and with ultimate pH (r = 0.708). It is concluded that high ultimate pH induced by stress significantly increases m-calpain activity, and this results in a greatly enhanced tenderisation of beef meat.     

The effect of time and type of electrical stimulation on the calpain system and meat tenderness in beef longissimus dorsi muscle

Effects of type and time of electrical stimulation on the calpain system, sensory and objective meat quality in the M. longissimus thoracis et lumborum from 38 pasture-fed carcasses, were investigated under conventional chilling conditions. High voltage stimulation was applied to whole carcasses at 3 min post-mortem (pm) and to sides at either 40 or 60 min pm, whilst low voltage stimulation was applied to whole carcasses at 3 min pm and to sides at 40 min pm. Unstimulated sides served as controls. The levels of extractable μ-calpain and calpastatin decreased during stimulation by 28-44% and 8-17%, respectively. Shear force and adjusted tenderness score showed that stimulation at 3 min, irrespective of type of stimulation, resulted in significantly tougher meat (P<0.05) which was associated with an rapid rate of pH decline, compared to stimulation at 40 min. Higher calpastatin levels soon after stimulation at 3 min (P < 0.05) and lower μ-calpain level at 24 h pm for high voltage stimulation at 3 min (P<0.05) coincided with the tougher meat. On the other hand, high voltage stimulation at 40 and 60 min resulted in similar tenderness and levels of μ-calpain and calpastatin post-stimulation and 24 h pm. Significantly tougher meat from the control sides, with a higher μ-calpain levels at 24 h pm and similar sarcomere length, compared to those from low voltage stimulation at 40 min (P<0.001), appeared to be linked to the later activation of the calpain system. Results from the current study suggest that early application of stimulation may be associated with a very rapid decline in pH and consequently a reduction in meat quality.     

Levels of calpain and calpastatin in meat subjected to high pressure

The levels of μ-, m-calpain and calpastatin were assayed in pressurized rabbit muscle.

The crude calpain level from the pressurized muscle at 100 MPa was almost the same with that of control. Above 100 MPa, the level of calpain decreased rapidly with increased pressure. At 300 MPa, the calpain level was almost inactivated. When the crude extract was pressurized, the calpain level followed the same tendency as that in the pressurized muscle.

When the extracts from control or pressurized muscle were subjected to DEAE-Sephacel column chromatography, μ- and m-calpains and calpastatin lost their activity with increasing pressure, but the degree of loss was different for each.

Calpains resisted changes in pressurization at 200 MPa and were inactivated over 200 MPa. Inactivation of calpastatin at 100 MPa was faster than that of calpains.

From the results, it was concluded that calpain levels remained in muscle pressurized up to 200 MPa, whereas calpastatin levels were decreased by the pressurization. Thus the total activities of calpains in pressurized muscle appear to have been increased by the pressure treatment and this may result in tenderization of meat.     

The interaction between pH and temperature decline early postmortem on the calpain system and objective tenderness in electrically stimulated beef longissimus dorsi muscle

The study investigated the effects of pH and temperature decline early postmortem (pm) on calpains and calpastatin activities and objective tenderness in M. longissimus thoracis et lumborum. A total of 40 sides were used to generate a range of pH and temperature declines, by different stimulation treatments (i.e. low voltage stimulation applied for either 10 or 40 s 3 min pm), and chilling treatments (fast or slow). Both rapid glycolysis, or a slow chilling regime significantly reduced μ-calpain level at 4 h pm (P< 0.001 and P< 0.05, respectively). There were significant interactions between pH and temperature at 1.5 h pm on μ-calpain and calpastatin activities at 24 h pm (P< 0.05). Rapid glycolysis resulted in lower shear force and compression measurements at day 1, although because of low ageing rates, an intermediate pH decline (i.e. pH 5.9-6.2 at 1.5 h pm) resulted in the most tender meat after 14 days ageing. Higher shear forces in the slow glycolysing sides appeared to be associated with the later activation of tenderising process, as well as physical shortening. The optimum pH decline early pm varied with days ageing, with an intermediate pH decline resulting in the lower shear forces for meat aged for 14 days.     

Perimysial collagen crosslinking and meat tenderness in Belgian Blue double-muscled cattle

The relationship between intramuscular collagen and five collagen crosslink concentrations, and the tenderness of meat from Belgian Blue normal, heterozygous double-muscled (DM) and homozygous DM cattle was investigated using M. semitendinosus (St) and M. gluteobiceps (Gb). The histidinohydroxymerodesmosine (HHMD) concentration (per mol collagen) in St was less in DM animals than normal animals. Concentrations (per gram of wet meat) of HHMD and Erlich chromogen (EC) in Gb, and HHMD, EC, dihydroxylysinorleucine (DHLNL) and hydroxylysinorleucine (HLNL) in St were also lower in DM animals than normal animals. Shear force of raw meat was significantly greater in normal animals than DM for both muscles; cooked meat shear force was greater in the normal animals for the Gb muscles only, showing a good correlation with sarcomere length. Most correlations between shear force and collagen or crosslink concentrations were not significant and those that were highly significant were generally weak.     

Post-mortem kinetics of meat tenderness and the components of the calpain system in bull skeletal muscle

Eight strip loins (M. longissimus dorsi) from pasture fed Friesian bulls were aged at 15 °C for a range of times from 1 to 120 h. pH declined from 6.29 (SE 0.119) one hour post slaughter to an ultimate pH of 5.48 (SE 0.013). The activities of the components of the calpain system (μ-calpain, m-calpain and calpastatin) were determined after separation on a DEAE-sephacel column. There was a dramatic decline in μ-calpain activity post slaughter with a complete disappearance within 48 h. The rates of decline in m-calpain and calpastatin activity were slower with 30% and 50% remaining 120 h post slaughter, respectively. The rapid decline in μ-calpain activity relative to the calpastatin activity is likely to reduce the degree of tenderisation and ultimate tenderness of the meat.     

The effect of altered growth rates on the calpain proteolytic system and meat tenderness in cattle

The present study was conducted to determine the effects of growth pattern on the calpain system and meat tenderization. Twenty-four Friesian calves were randomly allocated to three treatment groups: FAST (fast growth rate), SLOW (severely restricted growth rate) and ALTER (restricted growth for 30 days followed by fast growth rate). Four animals from each group were slaughtered on day 32 or 45 after altering the growth rates. Samples of M. longissimus dorsi were rapidly frozen at slaughter for protein analysis by Western blotting. Restricted growth reduced the immunoreactivity of a calpastatin band (135 kDa) measured at 24 h postmortem. Immunoreactivity associated with the large subunit of μ- or m-calpain appeared to be unaffected by growth patterns. Shear force measurements taken after 14 days of conditioning were positively related to 135 kDa calpastatin at 24 h postmortem. In this study there was no clear relationship between shear force and growth pattern.     

Effects of genetic variants for the calpastatin gene on calpastatin activity and meat tenderness in Hanwoo (Korean cattle)

This study was designed to investigate the effects of calpastatin genotypes determined by PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) on calpastatin activity (CAC) and Warner-Bratzler Shear Force (WBS). Longissimus muscles were prepared from 379 Hanwoo bulls aged approximately 20 months. The selection of PCR primers was based on exons (27 and 28) of the bovine calpastatin cDNA sequences, and genetic variants were detected by SSCP analysis using Taq I restriction enzymes. Sequencing analysis confirmed 4 restriction sites (nucleotide positions 52, 67, 796, and 1369), and a genetic variant was verified at a nucleotide position 641 (C/T substitutions) based on sequences (AF281256). The CAST28 genotypes showing allele frequencies of C (0.429) and T (0.571) were significantly associated with CAC and WBS. A significant positive residual correlation (r = 0.121, P = 0.02) between CAC and WBS was obtained.     

Modelling post-mortem tenderisation-IV: Role of calpains and calpastatin in conditioning

A generalised model, based on published data, was developed quantifying the mechanism by which the activities of calpains I and II are responsible for post-mortem tenderisation. Tenderisation is proposed to result from the activities of 'free' activated-calpains, the activities of which are controlled by the changes in the calcium ion concentration, the binding of calpains to calpastatin, the inactivations of 'free' activated-calpains and their proteolysis of calpastatin. At the low myoplasmic ('free') calcium ion concentrations prevailing soon after slaughter, calpains are largely 'inert' and little tenderisation occurs. As the pH declines, the 'free' calcium ion concentration rises and activates calpain I: however, most of this activated-calpain I is then bound to calpastatin. With a futher decline in pH, the binding of activated-calpain I to calpastatin is reduced and the level of 'free' activated-calpain I rises and tenderisation increases. A comparable process occurs with calpain II at higher 'free' calcium ion concentrations which occur as the pH declines further. As the level of 'free' activated-calpains rises, their proteolysis of calpastatin also increases, causing a lowering of levels of calpastatin and reducing the inhibition of calphins. Concurrently, 'free' activated-calpains are inactivated. This balance between inhibition, inactivation and activity of calpains and their decrease as the pH declines maintains the proteolytic activity of calpains and produces the gradual process of tenderisation, initially by calpain I and at the later stages mainly by calpain II. Eventually, the activities of calpains decline to zero and tenderisatin stops. Equations were derived to describe the changes from stunning to the completion of conditioning, and parameters were calculated to determine the activities of calpains and tenderisation in beef M. longissimus dorsi.     

Hind-limb protein metabolism and calpain system activity influence post-mortem change in meat quality in lamb

This study is concerned with the rate of protein turnover in the hind limb muscle bed of intact lambs, the activity of calpain proteolytic system in the M. biceps femoris, and subsequent rates of myofibre breakdown and tenderisation in the M. longissimus dorsi. Feed restriction increased protein degradation in hind-limb muscle of lambs (p<0.1), with a concominant decrease in the extractable activity of calpastatin (p<0.01), the endogenous inhibitor of calpain. IGF-1 analog treatment decreased both protein degradation and assayed μ-calpain activity (p<0.05) with no effect on the activity of calpastatin. β-Agonist treatment increased hind-limb protein synthesis (p<0.01), calpastatin activity (p<0.1) and decreased (p<0.01) μ-calpain activity, but did not effect protein degradation. Significant correlations were observed between Myofibril Fragmentation Index (MFI) values during post-mortem storage and initial post-slaughter calpastatin activity at days 3 (r=-0.34, p<0.1), 5 (r=-0.58, p<0.01) and 9 (r=-0.58, p<0.1), and μ-calpain activity at days 5 (r=0.35, p<0.1) and 9 (r=0.41, p<0.05). However, stronger correlations were observed between the ratio of μ-calpain to calpastatin, an estimate of potential μ-calpain proteolytic activity, and the rate of myofibril fragmentation (r=0.75, p<0.001) and tenderisation (r=-0.64, p<0.01) during aging. These results are consistent with the calpain system being the major proteolytic system involved in myofibril fragmentation and hence aging-related tenderisation of meat.     

Tensile strength and the tenderness of beef sternomandibularis muscle

In their relationship to shortening, the tensile strength of cooked meat along the fibres and the shearing force measured across them are strikingly similar. Both increase to maximum values at 40% shortening before falling by a half with an approach to 60% shortening. A high correlation (r = 0·81) exists between the two sets of values, whether toughness is increased by cold shortening or reduced by ageing. The mechanical strength of meat along its fibres is therefore a simple measure of tenderness and a useful basis for relating muscle structure to stength.     

Extensibility,strength and tenderness of beef cooked to various degrees

The response to loading of strips of ox sternomandibularis muscle varies greatly with degree of cooking. In raw or lightly cooked strips a yield point is reached where the myofibrils fail. Within this range, the yield point is independent of degree of cooking, and at 1.4 kg/cm² is well below the tension which can be developed in the pre-rigor muscle (2.3 kg/cm²). On cooking, the yield point vanishes between 10 and 40 min at 70°C and the strips behave more elastically. This degree of heating coincides with the sharpest change in shrinkage and other properties. By using strips in which the myofibrillar component has been destroyed by alkali, and others in which collagen has been destroyed by long cooking, an attempt has been made to separate the contributions of these components. Between 60 and 80°C a fall in connective tissue strength is matched by a rise in myofibrillar strength, maintaining a constant overall value of ca 5 kg/cm². Both components become more extensible and stretch in unison, until they fail together. After cooking at 100°C, only the myofibrillar component survives (ca 3 kg/cm²). Shear force values are generally in line with these results, but show a dip on cooking at 60°C, due to accelerated ageing. It is suggested that gap filaments may determine the tensile strength of the raw or cooked myofibril.