温度、pH对离体模型中肌原纤维蛋白去磷酸化反应的影响

摘要/Abstract

摘要： 蛋白质磷酸化修饰是最为普遍的一种翻译后修饰方式，其反应可逆，可经磷酸酶催化发生去磷酸化反应，碱性磷酸酶是肉中常见的磷酸酶。已有研究表明畜禽宰后肉中存在多种磷酸化调节通路，且蛋白质磷酸化负向影响肉嫩度、色泽等品质。本研究旨在探究不同温度（25℃、15℃、4℃）和pH（5.2、5.8、6.4）对碱性磷酸酶活性及催化去磷酸化反应能力的影响，为改善肉品质提供理论依据。提取宰后羊肉中的肌原纤维蛋白建立离体模型，添加碱性磷酸酶催化肌原纤维蛋白发生去磷酸化反应。采用SDS-PAGE凝胶电泳、Pro-Q Diamond磷酸化蛋白染色和SYPRO Ruby全蛋白染色测定肌原纤维蛋白磷酸化水平，试剂盒测定碱性磷酸酶活性。结果表明，随着孵育时间延长，pH 5.8和pH 6.4条件下碱性磷酸酶组肌原纤维蛋白磷酸化水平显著下降（P < 0.05），pH 5.2条件下碱性磷酸酶组肌原纤维蛋白磷酸化水平无显著变化（P > 0.05）。对照组肌原纤维蛋白磷酸化水平在整个孵育期间无显著变化（P > 0.05）；4℃、pH 5.2条件下显著抑制（P < 0.05）碱性磷酸酶活性，使其催化去磷酸化时间延迟。测定碱性磷酸酶磷酸化水平发现，碱性磷酸酶不仅可以催化肌原纤维蛋白发生去磷酸化反应，也能催化其自身发生去磷酸化反应，且碱性磷酸酶自身发生去磷酸化后不会失活。因此在一定范围内升高温度和pH会提高碱性磷酸酶催化底物去磷酸反应的能力，降低磷酸化水平，且使去磷酸化时间提前，为改善肉品质及肉贮藏提供新思路。

关键词: 蛋白质磷酸化, 碱性磷酸酶, 去磷酸化, 温度, pH

Abstract: Protein phosphorylation is a universal posttranslational modification, and the reaction of protein phosphorylation is reversible. Under phosphatase catalyzing, protein will happen dephosphorylation. Alkaline phosphatase (AP) is a common phosphatase in postmortem muscle. Furthermore, different regulating pathways of phosphorylation on meat quality have been reported by previously studies, and phosphorylation negatively affects meat quality such as tenderness and color. This study mainly determined the effect of temperature (25℃, 15℃, 4℃) and pH (5.2, 5.8, 6.4) on AP activity and dephosphorylation of myofibrillar protein, which will provide theoretical basis for improving meat quality. Myofibrillar protein of postmortem muscle was extracted and AP was added to catalyze myofibrillar protein dephosphorylate. The phosphorylation levels were detected by using SDS-PAGE and gels were stained by Pro-Q Diamond and SYPRO Ruby. Moreover, AP activity was measured by kits. According to the results, under pH 5.8 and pH 6.4, the myofibrillar protein phosphorylation levels of AP group were significantly decreasing (P < 0.05). However, the myofibrillar protein phosphorylation levels of AP group were no significant difference (P > 0.05) when incubated at pH 5.2. That result of control groups were not significantly changes all the time (P > 0.05). When incubated at 4℃ and pH 5.2, the AP activity were restrain delayed catalyzing time (P < 0.05). Interestingly, AP not only could catalyze myofibrillar protein dephosphorylation, but also could catalyze itself dephosphorylate. The AP activity wouldn’t be inactivated after catalyzing itself dephosphorylate. Suitably, the increasing temperature and pH would promote dephosphorylation ability of AP which decreased phosphorylation levels and advanced catalyzing time. These foundings played an important role in improving meat quality and provided a new idea to store postmortem muscle.

Key words: protein phosphorylation, alkaline phosphatase, dephosphorylation, temperature, pH

中图分类号:

TS251.1

任驰侯成立李欣摆玉蔷张德权. 温度、pH对离体模型中肌原纤维蛋白去磷酸化反应的影响[J]. 食品科学, 0, (): 0-0.

Xin Li. Effect of temperature and pH on dephosphorylation of myofibrillar protein in vitro[J]. FOOD SCIENCE, 0, (): 0-0.

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