贻贝粘蛋白中多巴氧化还原状态表征方法研究

Study on the 3,4-Dihydroxyphenylalanine Redox State Characterization Method of Mussel Adhesive Protein

摘要: 目的对高效液相色谱法表征贻贝粘蛋白中多巴氧化-还原状态可行性进行研究。方法用高效液相色谱法、Arnow法和考马斯亮兰法测定贻贝粘蛋白中多巴含量及蛋白质含量。结果随着氧化时间延长,贻贝粘蛋白样品中蛋白含量不变,还原态的多巴含量下降。加速氧化和自然存放样品的色谱峰保留时间均随着放置时间的延长发生偏移,与多巴的氧还还原状态变化有关。结论高效液相色谱法可以用来表征贻贝蛋白的氧化还原程度,对于产品设计研发与质量控制具有重要意义。

Abstract: Objective To investigate the feasibility of using liquid chromatography (HPLC) to characterize the 3, 4-Dihydroxyphenylalanine (DOPA) redox state of mussel adhesive protein (MAP).Methods The DOPA and protein contents of MAP were determined by HPLC, Arnow and Bradford methods respectively.Results With extended oxidation time, the protein contents of MAP samples remained unchanged whereas the DOPA contents declined. The retention times of main peaks in HPLC for both the accelerated oxidation and retained samples shifted as the storage time extended, which could be related to the changes of sample redox state.Conclusion The redox state of MAP can be characterized by the change of HPLC peak retention time. HPLC can be used in the research on the MAP redox state, which is beneficial to the product development and quality control.