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[目的]纯化源自生防细菌K2-1的抗菌蛋白并进行抗菌蛋白于体外对主要水产病原菌的拮抗分析。[方法]利用硫酸铵沉淀,而后透析,获得抗菌蛋白粗品,应用凝胶色谱结合水浴,进一步对抗菌蛋白粗品进行纯化;采用琼脂扩散法,进行抗菌蛋白体外拮抗溶藻弧菌、嗜水气单胞菌、温和气单胞菌、荧光假单胞菌、副溶血弧菌、哈维氏弧菌及鳗弧菌实验,分析其抗菌活性。[结果]应用上述分离纯化方法,获得色谱纯的抗菌蛋白APK2,过程回收率为0.08%,拮抗实验表明,该抗菌蛋白对多数应试病原菌具有较强的抑制作用。[结论]抗菌蛋白APK2有可能作为化学抗生素的替代品,用于水产细菌病的绿色防治。
[Objective] The research aimed to purify the antibacterial protein from biocontrol bacterium K2-1 and conduct the antagonistic analysis of antibacterial protein against the major aquatic pathogenic bacteria in vitro. [Method] The precipitate of antimicrobial protein was obtained by precipitation with ammonium sulfate and dialysis to obtain crude antibacterial protein. The crude product was further purified by gel chromatography and water bath. The agar diffusion method was used to antagonize Vibrio alginolyticus, Aeromonas sobria, Aeromonas sobria, Pseudomonas fluorescens, Vibrio parahaemolyticus, Vibrio harveyi and Vibrio anguillarum. The antibacterial activity was analyzed. [Result] The chromatographic pure antibacterial protein APK2 was obtained by the above purification method. The recovery rate was 0.08%. The antagonistic experiment showed that the antibacterial protein had a strong inhibitory effect on most pathogenic bacteria. [Conclusion] The antimicrobial protein APK2 may be used as a substitute for chemical antibiotics for the green prevention and control of aquatic bacterial diseases.