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Meat Tenderness and the Calpain Proteolytic System in Longissimus Muscle of Young Bulls and Steers1

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Meat Tenderness and the Calpain Proteolytic System in Longissimus Muscle of Young Bulls and Steers1 Meat Tenderness and the Calpain Proteolytic System in Longissimus Muscle of Young Bulls and Steers1 J. B. Morgan**2, T. L. Wheeler?, M. Koohmaraie', J. W. Savell", and J. D. CrousetJ +Roman L. Hruska U.S. Meat Animal Research Center, ARS, USDA, Clay Center, NE 68933-0166 and *Department of Animal Science, Texas A&M University, College Station 77843-2471 ABSTRACT: The objectives of this study were to than meat from steers; however, sensory panelists examine the effects of castration on the calpain were unable (P > .05) to detect differences in proteinase system ( p-calpain, m-calpain, and cal- tenderness or other sensory traits between bulls and pastatin) activities and meat tenderness. Six each, steers. Activities of p- and m-calpain were not affected MARC 111 bulls and steers were slaughtered at (P > .05) by castration; however, calpastatin was approximately 12 mo of age. Longissimus muscle higher (P c .05) in muscles from the bull carcasses. samples were obtained for determining myofibril fragmentation index, Warner-Bratzler shear force, and Lower ( P c .05) myofibril fragmentation index values sensory panel evaluation at 1, 7, and 14 d postmortem, indicate that less proteolysis occurred in muscle from and p- and m-calpain and calpastatin activities at 24 h bulls than in muscle from steers during the first 7 d postmortem. Bulls produced leaner carcasses with postmortem. Greater calpastatin 24-h activity may be lower (Pc .05) quality grades than did steers. Meat associated with the increased shear force of meat from from bulls had higher ( P c .05) shear force values bulls. Key Words: Beef, Calpains, Calpastatin, Castration, Meat, Tenderness J. him. Sci. 1993. 71:1471-1476 Introduction 1986; Dikeman et al., 1986). The mechanism causing tenderness differences between bulls and steers is not Numerous research studies have been conducted to well defined. Numerous studies have attempted to assess growth and meat characteristic differences link bull meat toughness to higher amounts of between bulls and steers. In general, results have connective tissue than that of steers (Riley et al., indicated that bulls grow more rapidly (15 to 17%), 1983; Grouse et al., 1985; Vanderwert et al., 1986). utilize feed more efficiently (10 to 13%) to an age or However, a strong relationship between 24-h postmor- weight end point, and produce higher yielding car- tem calpastatin activity, myofibrillar proteolysis, and casses with less fat and more muscle than steers meat tenderization has been reported in steers and (Field, 1971; Seideman et al., 1982). However, in heifers (Whipple et al., 1990; Koohmaraie and Shack- most cases, meat from young bulls was more variable elford, 1991; Shackelford et al., 1992). The possible in tenderness than meat from steers (Burson et al., contribution of the calpain proteinase system to differences in postmortem tenderization of muscle from bulls and steers has not been reported. This lThe technical assistance of P. Ekeren and P. Gilmore is study was conducted to examine the effect of castra- gratefully acknowledged by the authors. Names are necessary to tion on palatability traits and 24-h postmortem report factually on available data; however, the USDA neither activities of p-calpain, m-calpain, and calpastatin of guarantees nor warrants the standard of the product, and the use of longissimus muscle in cattle. the name by USDA implies no approval of the product to the exclusion of others that may also be suitable. Technical Article no. 30570 from the Texas Agric. Exp. Station. Partial funding was provided by King Ranch, Inc. to J. B. Morgan and J. W. Savell. Materials and Methods 2Present address: Dept. of Anim. Sci., Colorado State Univ., Fort Collins, CO 80523. Animals. The Roman L. Hruska U.S. Meat Animal 'Present address: USDA, ARS, NPA, 1201 Oakridge Drive, Fort Collins, CO 80525-5562. Research Center Animal Care and Use Committee Received July 30, 1992. approved the use of animals in this study. Animals to Accepted January 15, 1993. remain bulls or to be castrated were randomly 1471 1472 MORGAN ET AL. assigned and castration was performed within 1 wk of from 25 to 350 mM NaC1. Activities were expressed as birth. Six each, MARC I11 composite (1/4 Red Poll, 1/4 the amount of calpain caseinolytic activity per gram of Pinzgauer, 114 Hereford, and 1/4 Angus) bulls and muscle. One unit of p- and m-calpain activity was steers weighing approximately 180 kg at approxi- defined as the amount of enzyme that catalyzed an mately 60 mo of age were given access to a diet increase of 1.0 absorbance unit at 278 nm in 1 h at formulated to meet NRC requirements for growing 25°C. One unit of calpastatin activity was defined as beef cattle (NRC, 1984). The diet consisted of 36% the amount that inhibited 1.0 unit of DEAE-Sephacel corn silage, 60% wet corn, and 4% supplement purified m-calpain activity. (supplement composition: 54% soybean meal, 17.9% Shear Force and Sensory Evaluation. Steaks were corn, 21.7% limestone, 2% dicalcium phosphate, 3% tempered at 2°C for 24 h before cooking. The steaks urea, .6% vitamin A, .2% trace minerals, 5% Rumen- were broiled to an internal temperature of 40"C, sin [containing 132 g of monensinkg], and .l% sulfur) turned, and broiled to an internal temperature of 70°C with a TDN of 84.16% and 10.93% CP. Animals were on Farberware electric broilers (Farberware, Bronx, weighed weekly. The experiment was conducted from NY) for determination of shear force and sensory November through May. Animals were housed in an panel analysis at 1, 7, and 14 d postmortem. The insulated barn (temperature maintained at approxi- internal temperature was monitored by a copper mately 13°C) in individual stalls. Lights in the constantan thermocouple probe attached to a Honey- building were on 9 h and off 15 h during each well recording thermometer (Honeywell, Scarborough, 24-h period. Animals were removed from the stalls ON, Canada). Weights were recorded before and after and allowed to exercise outside for 3 h, twice weekly. cooking for determination of percentage of cooking Animals were fed for 168 d and slaughtered according loss. Steaks were allowed to cool for 2 h at 23"C, and to standard humane procedures at approximately 12 10 1.3-cm cores were removed from each steak parallel mo of age. Carcasses were chilled for 24 h at 1°C. to the longitudinal orientation of the muscle fibers. Carcass Evaluation. At 24 h postmortem, the left Each core was sheared once with a Warner-Bratzler side of each carcass was ribbed between the 12th and shear machine. Shear force for each cut was recorded 13th ribs for determination of USDA quality and yield as the mean of the 10 cores. grade factors (USDA, 1989). Hot carcass weight, Steaks for sensory evaluation were cooked as dressing percentage, adjusted fat thickness, marbling described above. Warm l-cm3 samples were served to degree, longissimus muscle area, and percentage of an eight-member sensory panel trained according to kidney, pelvic, and heart fat were recorded. Dark, methods described by Cross et al. (1978). The panel coarse band formation (heat ring), lean color, lean evaluated each sample for juiciness, myofibrillar and firmness, and lean texture were scored on a overall tenderness, connective tissue amount, and 7-point scale (7 = severe, black, very soft, or very flavor intensity based on %point scales (8 = extremely coarse, and 1 = none, very light cherry red, very firm, juicy, tender, none, and intense; 1 = extremely dry, or very fine). tough, abundant, and bland). At 24 h postmortem, the longissimus muscle (right Myofibril Fragmentation Index. Samples were ex- side) from the 7th thoracic to the 5th lumbar vertebra cised from the longissimus muscle at 24 h postmortem was cut into steaks 2.5-cm thick and vacuum- and were vacuum-packaged and stored for an addi- packaged. Two steaks each were assigned to 1, 7, or 14 tional 6 or 13 d at 4°C. Myofibril fragmentation index d postmortem aging (2"C) by stratifying storage time ( MFI) was determined on fresh muscle at 1, 7, and 14 along the length of the longissimus muscle. One steak d postmortem according to the procedures of Culler et was used for Warner-Bratzler shear force and one for al. (1978). The biuret procedure was used to deter- sensory panel evaluation. The steaks were stored at mine protein concentrations (Gornall et al., 1949). -10°C for 2 to 4 wk until they were thawed and Statistical Analysis. Data were analyzed by analysis cooked. of variance with the GLM procedure of SAS ( 1985) for Calpain and Calpastatin Determination. At 24 h a split-plot design. The whole plot was animal sex postmortem, a 5-g sample of longissimus muscle was class (bull vs steer) and the split plot was postmortem taken from one of the shear force steaks from the 12th aging time (Warner-Bratzler shear, MFI, sensory rib region of the right side of each carcass for traits, and cooking loss). The whole-plot error term in determinations of p-calpain, m-calpain, and calpasta- the model was replication x sex class with animal tin activities according to the procedure described by representing replication. The split-plot error term in Wheeler and Koohmaraie (1991) using 50 mM Tris- the model was the residual error. Tukey's test was HC1, pH 8.3, instead of 50 mM sodium acetate, pH 5.8, used for mean separation when needed. as the extraction solution. Briefly, after homogeniza- tion, centrifugation, dialysis, and clarification, the muscle extracts were loaded onto DEAE-Sephacel Results and Discussion (Pharmacia LKB, Piscataway, NJ) columns and washed with 10 column volumes of elution buffer to Live performance traits of animals in this study remove unbound proteins.
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