title: Indicators of tenderization are detectable by 12 h postmortem in ovine longissimus.
abstract: Postmortem changes in osmotic pressure; ionic strength; pH; temperature; mu- and m-calpain; calpastatin; desmin degradation; and myofibril fragmentation index (MFI) were determined in ovine longissimus muscle. Our objectives were to characterize changes in these variables and to identify postmortem time points at which significant proteolysis and tenderization (as measured by change in MFI) could be detected. Seven crossbred (Dorset x Romanov) lambs were slaughtered, and samples of the longissimus muscle were removed at 0, 3, 6, 9, 12, 24, 72, and 360 h postmortem. Osmotic pressure increased (P < 0.05) from 379 to 528 mOsm during the postmortem storage period, with two-thirds of the increase occurring within the first 24 h. By measuring conductivity, we showed that ionic strength increased (P < 0.05) from 8.13 to 9.78 mS/cm during the storage period, which is equivalent to 79 and 97 mM NaCl solutions, respectively. In accordance with pH and temperature, conductivity reached ultimate levels at 24 h postmortem. Within 9 h postmortem, mu-calpain activity had decreased (P < 0.05) from at-death values and continued to decrease until 72 h, at which time it was undetectable. It was still possible to detect the 76-kDa isoforms (a product of the autolysis of the 80-kDa subunit of mu-calpain) immunologically, which implies that the loss of activity was not caused by extensive autolysis. In contrast, m-calpain activity remained constant throughout the aging period, whereas calpastatin activity was stable until 24 h postmortem, after which it gradually decreased. Autolysis products of mu-calpain were detected at 3 h postmortem, indicating that mu-calpain was activated some time between 0 and 3 h postmortem. Moreover, the effect of mu-calpain activity on myofibrillar substrates was first observed at 9 h postmortem, when a 23% loss of native desmin was detected. This degradation translated into an increase in MFI at 12 h. Collectively, these results imply that mu-calpain is active in postmortem muscle in the presence of calpastatin, and that effects of mu-calpain activity as determined by increased MFI are detectable during the first 12 h postmortem.
authors.affiliation: Department of Chemistry, Biotechnology, and Food Science, Agricultural University of Norway, 1432 Aas, Norway.
authors.forename: E
authors.initials: E
authors.lastname: Veiseth
authors.forename: S D
authors.initials: SD
authors.lastname: Shackelford
authors.forename: T L
authors.initials: TL
authors.lastname: Wheeler
authors.forename: M
authors.initials: M
authors.lastname: Koohmaraie
mesh_terms: D000818:Animals; D001831:Body Temperature; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002154:Calpain / Q000378:metabolism; D003893:Desmin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D008460:Meat / Q000592:standards*; D018482:Muscle, Skeletal / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism; D009994:Osmolar Concentration; D009997:Osmotic Pressure; D011180:Postmortem Changes; D012756:Sheep; D013997:Time Factors
chemical_list: D002135:Calcium-Binding Proteins; D003893:Desmin; C033668:calpastatin; D002154:Calpain