原核6-4 光修复酶中天线辅酶类型的研究  

Types of antenna coenzyme in prokaryotic 6-4 photolyase

作　　者：张薇 吕俊 尚雪 周鸿铭 徐蕾 ZHANG Wei;L Jun;SHANG Xue;ZHOU Hongming;XU Lei(Anhui Provincial Key Laboratory of Active Biological Macro-molecules Research,Wannan Medical College,Wuhu 241002,Anhui,China)

机构地区：[1]皖南医学院活性生物大分子研究安徽省重点实验室,安徽芜湖241002

出　　处：《皖南医学院学报》2025年第2期112-115,共4页Journal of Wannan Medical College

基　　金：国家自然科学基金面上项目(32271287,31971199);皖南医学院科学研究“揭榜挂帅”专项(WK2022J03);活性生物大分子研究安徽省重点实验室校级开放课题(LAB202205)。

摘　　要：目的:鉴定及转换原核6-4光修复酶中结合天线辅酶的类型,预测天线辅酶结合存在的进化规律并增加光修复效率。方法:突变关键位点的氨基酸残基,在大肠杆菌中异源表达及纯化相关蛋白,通过紫外吸收光谱和薄层层析的方法,检测并鉴定了天线辅酶的类型。结果:鉴定了来自B类的禽波氏杆菌结合的天线辅酶为DMRL,来自Ex类的微小杆菌结合8-HDF作为其天线辅酶,并且成功将农杆菌原核6-4光修复酶原始所结合的天线辅酶DMRL转换为8-HDF。结论:8-HDF作为天线辅酶可能是含天线辅酶光修复酶进化过程中的祖先,在酶的高效改造和临床应用方面发挥作用。Objective:To identify and transform the types of antenna binding coenzymes in prokaryotic 6-4 photolyase,predict the evolution of antenna coenzyme binding,and increase the efficiency of photorepair.Methods:Amino acid residues of key sites were mutated,and the related proteins were heterologously expressed and purified in Escherichia coli.The types of antenna coenzymes were detected and identified by UV absorption spectroscopy and thin layer chromatography.Results:DMRL was identified as the antenna coenzyme bound by Bordeella avium from class B,and 8-HDF was bound by Exiguobacterium from class Ex,and the antenna coenzyme DMRL originally bound to 6-4 photolyase of Agrobacterium fabrum was successfully converted to 8-HDF.Conclusion:As an antenna coenzyme,8-HDF may represent an ancestral form in the evolution of photolyase containing antenna coenzyme,and functions a significant role in the efficient engineering and clinical application of enzymes.

关 键 词：紫外损伤 光修复酶 辅酶转换 进化 

分 类 号：Q503[生物学—生物化学] R818[医药卫生—放射医学]