大鲵皮肤水溶性蛋白质成分分析及活性鉴定

Composition Analysis and Activity Identification of Skin Soluble Protein of Andrias davidianus

该试验旨在研究大鲵(Andrias davidianus)皮肤水溶性蛋白成分及其活性,揭示大鲵皮肤黏液的蛋白组分,鉴定具有潜在功能活性的蛋白质。通过物理和化学刺激的方法收集大鲵皮肤黏液,制备大鲵皮肤黏液水溶性蛋白样品,并通过SDS-PAGE垂直电泳、细菌凝集试验、氧化物清除效率试验以及蛋白质谱Label-free分析大鲵皮肤黏液水溶性蛋白的成分以及活性。结果表明:大鲵皮肤黏液中含有多种水溶性蛋白,分子量介于10~300 kDa之间,其中50 kDa分子量以上蛋白条带较为丰富,在100 kDa分子量以上含有较多蛋白,14 kDa以下蛋白含量较少。大鲵皮肤黏液水溶性蛋白对1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABST)2种自由基具有清除能力,说明其具有一定的抗氧化活性。大鲵皮肤黏液水溶性蛋白对大肠杆菌和嗜水气单胞菌2种细菌具有较强的凝集作用,暗示大鲵皮肤水溶性蛋白具有一定的抗菌活性。蛋白质谱Label-free分析表明这些蛋白是由66种蛋白质组成;通过分子注释有20种蛋白与免疫抗病功能相关,2种蛋白与抗氧化活性相关。研究揭示大鲵皮肤黏液水溶性蛋白含有66种蛋白,分子量介于10~300 kDa,活性鉴定结果显示大鲵黏液水溶性蛋白具有抗氧化活性和抗菌活性。

To study the components and activities of water-soluble proteins of the skin mucus in Chinese giant salamander(Andrias davidianus)and provide a reference for the further development of biological resources of skin mucus,in this study,the skin mucus was collected by physical and chemical stimulation methods to prepare samples of water-soluble proteins.The composition and activity of water-soluble proteins in the skin mucus of the salamander were analyzed by SDS-PAGE vertical electrophoresis assay,bacterial agglutination assay,oxide removal efficiency assay,and Label-free protein spectrum.Results showed the mucus of Chines giant salamander contained various water-soluble proteins,with molecular weight ranging from 10~300 kDa.Among them,protein bands for molecular weight above 50 kDa were more abundant,that above 100 kDa had more proteins,and that below 14 kDa contained less protein.The water-soluble protein from the skin mucus of Andrias davidianus has the scavenging ability to 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)and 2,2-binazo-bis(3-ethyl-benzothiazole-6-sulfonic acid)diamammonium salt(ABST),indicating that it has certain antioxidant activity.Besides,the skin mucus water-soluble protein of Andrias davidianus has strong agglutination effect on Escherichia coli and Aeromonas hydrophila,suggesting that the skin water-soluble protein of Salamander has certain antibacterial activity.Label-free analysis of protein spectrum showed that these proteins are composed of 66 proteins,and 20 proteins are related to immune resistance,and 2 proteins are related to antioxidant activity through molecular annotation.The water-soluble proteins in the skin mucus of the salamander contain 66 kinds of proteins with molecular weights ranging from 10-300 kDa.The results of activity identification suggest that the water-soluble proteins in the skin mucus of the salamander have antioxidant and antibacterial activities.