基于宰后蛋白质磷酸化研究L-组氨酸和L-赖氨酸对鸡胸肉加工特性的影响

Effect of L-Histidine and L-Lysine on the Processing Characteristics of Chicken Breast Meat Evaluated from the Perspective of Post-Slaughter Protein Phosphorylation

为研究低盐(1%NaCl)条件下添加L-组氨酸(L-histidine,L-His)及L-赖氨酸(L-lysine,L-Lys)调节宰后鸡胸肉肌原纤维蛋白磷酸化的变化规律及其对鸡胸肉加工特性(蒸煮损失、质构、蛋白溶解度、流变特性)的影响,提取腌制后鸡胸肉肌原纤维蛋白,进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,再通过荧光染色和液相色谱-串联质谱鉴定。结果表明:L-His和L-Lys的添加不影响肌原纤维蛋白的整体磷酸化水平,但是可以改变单个蛋白质的磷酸化水平;相比于1%NaCl处理组,1%NaCl+0.06%L-His/L-Lys处理组硬度和蛋白溶解度均有显著提高,蛋白流变特性改善,蒸煮损失无显著变化;相关性分析得出,多个蛋白质磷酸化水平与肉的蒸煮损失及蛋白溶解度等加工特性呈显著正相关或负相关,且单个蛋白质的磷酸化水平还会受到其他蛋白质磷酸化水平的影响;将11条磷酸化水平差异显著的条带进行质谱鉴定后,发现大多是与肌肉收缩或糖酵解有关的酶,表明L-His、L-Lys的添加可能通过对肌原纤维蛋白磷酸化水平产生正面效应影响宰后僵直成熟过程,进而影响肉品品质。

The effect of adding L-histidine(L-His)and L-Lysine(L-Lys)under the condition of low salt(1%sodium chloride,NaCl)on the phosphorylation of chicken breast myofibrillar protein after slaughter and the processing characteristics(cooking loss,texture,protein solubility and rheological properties)of chicken breast meat was studied by sodium dodecyl sulfatepolyacrylamide gel electrophoresis(SDS-PAGE),fluorescence staining and liquid chromatography tandem mass spectrometry(LC-MS/MS).The results showed that the addition of L-His and L-Lys did not affect the overall phosphorylation level of myofibrillar proteins,but could change the phosphorylation levels of individual proteins.Compared to the 1%NaCl-treated group,1%NaCl+0.06%L-His/L-Lys treatment significantly increased the heat-induced gel hardness and protein solubility,and improved the rheological properties of chicken myofibrillar protein during heat-induced gelation without having a significant effect on the cooking loss.Correlation analysis showed that the phosphorylation levels of multiple myofibrillar protein fractions were significantly positively or negatively correlated with the cooking loss and protein solubility,and that the phosphorylation levels of individual proteins were also influenced by those of other protein components.A total of 11 bands with significantly differential phosphorylation levels were identified,revealing that they were mostly enzymes related to muscle contraction or glycolysis.It was suggested that the addition of L-His and L-Lys may affect the post-slaughter stiffening and aging process through a positive effect on the phosphorylation level of myogenic fibronectin,which in turn affects meat quality.