Expressão das proteínas calpaína e calpastatina e suas relações com a qualidade da carne de bovinos da raça Nelore (Bos indicus)
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Data
2016-08-02
Autores
Dias, Victor Augusto Domingos [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A atuação das enzimas proteolíticas durante o post-mortem é um dos principais fatores que determinam a qualidade da carne, sendo as enzimas calpaína e calpastatina uma das principais atuantes neste processo. O objetivo deste estudo foi avaliar a relação entre a expressão gênica dos genes CAPN1, CAPN2 e CAST, a quantificação das enzimas µ-calpaína, m-calpaína e calpastatina, dentro de grupos contrastantes para maciez e crescimento animal. Foram utilizados dados de 90 animais machos não castrados da raça Nelore, com idade aproximada de 24 meses, permanecendo em confinamento por 95 dias. Após o abate foram colhidas amostras do músculo Longissimus thoracis entre a 9ª e 13ª costelas, da meia-carcaça esquerda. As amostras coletadas foram utilizadas para medir a área de olho de lombo (AOL), espessura de gordura subcutânea (EGS), índice de marmorização (IM), perdas por cozimento, coloração instrumental da carne e força de cisalhamento (FC). A partir do restante da carne, foram coletadas alíquotas para a realização das análises de índice de fragmentação miofibrilar (MFI) e lipídeos totais (LT). Foram coletadas alíquotas para a análise da expressão gênica e quantificação das enzimas. Os dados de força de cisalhamento foram utilizados como critério para a separação em carne macia e dura. Também foram utilizadas o ganho de peso para a separação quanto ao crescimento animal. Os seguintes grupos (n=10) foram formados: Leve, Pesado, Duro e Macio. As análises estatísticas foram realizadas utilizando-se o procedimento GLM e MIXED do programa SAS. No presente trabalho não foi encontrada diferença nas expressões dos genes CAPN1 e CAPN2 entre os grupos. Para o grupo contrastante para maciez também não foi encontrada diferença significativa (p<0,05 para a quantidade das enzimas µ e m-calpaína, e da calpastatina. Não foi encontrada correlação da quantidade das enzimas µ e m-calpaína, e calpastatina com as características PF e GP. Não foi encontrada diferença significativa (p<0,05) para o grupo contrastante para maciez da quantidade das enzimas µ e m-calpaína. Porém foi encontrada diferença significativa (p<0,05) para a quantidade da enzima calpastatina, onde os animais do grupo de carne dura apresentaram maiores valores em relação aos de carne macia. Além de apresentarem uma alta correlação entre a quantidade de calpastatina e as características FC (0) e FC (7) (r= 0,81 e 0,74, respectivamente). A diferença entre os grupos contrastantes e a alta correlação entre os níveis de calpastatina com as características de maciez demonstra a importância do sistema das calpainas na proteólise das miofibrilas, sendo os níveis de calpastatina um alvo potencial para a seleção de animais, com o objetivo de melhorar as características da qualidade da carne dos animais da raça Nelore.
The role of proteolytic enzymes during the post-mortem is one of the main factors that determine the quality of the meat, and the enzymes calpain and calpastatin are one of the main factors in this process. The aim of this study was to evaluate the gene expression of the genes CAPN1, CAPN2 and CAST, and quantify the enzymes μ-calpain, m-calpain and calpastatin within contrasting groups animal growth, and thougness. 90 animals data were used uncastrated male Nellore, aged approximately 24 months remaining in confinement for 95 days. After slaughter were harvested Longissimus muscle samples thoracis between the 9th and 13th ribs, the left half-carcase. The samples were used to measure the ribeye area (REA), subcutaneous fat thickness (SFT), marbling index (MI), cooking loss, instrumental color of the meat and shear force (SF). From the rest of the meat, aliquots were collected to perform the myofibrillar fragmentation index analysis (MFI) and total lipids (TL). Were collected aliquots for the analysis of gene expression and quantification of enzymes. The data of shear force were used as criteria for the separation of tender and tough meat. The characteristics of weight gain for the separation on the animal growth were also used. The following groups (n=10) were formed: lightweight, Heavy, tough meat and tender meat. Statistical analyzes were performed using the GLM and MIXED procedure of Statistical Analysis System program. In this study there was no difference in the expression of CAPN1 and CAPN2 genes between the groups. In this study there was no difference in the expressions of CAPN1 and CAPN2 genes between the groups. For the contrasting group for tenderness meat was also no significant difference (p<0.05) for the amount of enzymes μ and m-calpain, and calpastatin. There was no correlation between the amount of μ and m-calpain enzymes, and calpastatin with PF and GP. There was no significant difference (p<0.05) for the contrasting group for meat tenderness of the amount of enzyme μ and mcalpain. However, a significant difference (p<0.05) for the amount of enzyme calpastatin where animals of tough meat group had higher values on the tender meat. In addition to having a high correlation between the amount of characteristics calpastatin and SF (0) and SF (7) (r = 0.81 and 0.74, respectively). The difference between the contrasting groups and the high correlation between the calpastatin levels with tenderness characteristics demonstrates the importance of the system of calpains proteolysis of myofibrils, and the calpastatin levels of a potential target for the selection of animals for the purpose of improve the quality characteristics of the meat of Nellore.
The role of proteolytic enzymes during the post-mortem is one of the main factors that determine the quality of the meat, and the enzymes calpain and calpastatin are one of the main factors in this process. The aim of this study was to evaluate the gene expression of the genes CAPN1, CAPN2 and CAST, and quantify the enzymes μ-calpain, m-calpain and calpastatin within contrasting groups animal growth, and thougness. 90 animals data were used uncastrated male Nellore, aged approximately 24 months remaining in confinement for 95 days. After slaughter were harvested Longissimus muscle samples thoracis between the 9th and 13th ribs, the left half-carcase. The samples were used to measure the ribeye area (REA), subcutaneous fat thickness (SFT), marbling index (MI), cooking loss, instrumental color of the meat and shear force (SF). From the rest of the meat, aliquots were collected to perform the myofibrillar fragmentation index analysis (MFI) and total lipids (TL). Were collected aliquots for the analysis of gene expression and quantification of enzymes. The data of shear force were used as criteria for the separation of tender and tough meat. The characteristics of weight gain for the separation on the animal growth were also used. The following groups (n=10) were formed: lightweight, Heavy, tough meat and tender meat. Statistical analyzes were performed using the GLM and MIXED procedure of Statistical Analysis System program. In this study there was no difference in the expression of CAPN1 and CAPN2 genes between the groups. In this study there was no difference in the expressions of CAPN1 and CAPN2 genes between the groups. For the contrasting group for tenderness meat was also no significant difference (p<0.05) for the amount of enzymes μ and m-calpain, and calpastatin. There was no correlation between the amount of μ and m-calpain enzymes, and calpastatin with PF and GP. There was no significant difference (p<0.05) for the contrasting group for meat tenderness of the amount of enzyme μ and mcalpain. However, a significant difference (p<0.05) for the amount of enzyme calpastatin where animals of tough meat group had higher values on the tender meat. In addition to having a high correlation between the amount of characteristics calpastatin and SF (0) and SF (7) (r = 0.81 and 0.74, respectively). The difference between the contrasting groups and the high correlation between the calpastatin levels with tenderness characteristics demonstrates the importance of the system of calpains proteolysis of myofibrils, and the calpastatin levels of a potential target for the selection of animals for the purpose of improve the quality characteristics of the meat of Nellore.
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Palavras-chave
Proteólise, Qualidade da carne, Western blot, Proteolysis, Meat quality