Yin Manman, Liu Dan, Xu Feng, Xiao Liang, Wang Qianqian, Wang Beilei, Chang Yinlong, Zheng Jiemin, Tao Xia, Liu Guoyan, Zhang Liming
Marine Bio-pharmaceutical Institute, Second Military Medical University, Shanghai 200433, China; Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai 200433, China.
Department of Food Microbiology, College of Food Science and Engineering, Dalian Ocean University, Dalian 116021, China.
Int J Biol Macromol. 2016 Jan;82:488-96. doi: 10.1016/j.ijbiomac.2015.10.056. Epub 2015 Oct 31.
A bacterium strain, designated as CMF-2, was isolated from the jellyfish Cyanea capillata and its culture supernatant exhibited a significant antimicrobial activity. The strain CMF-2 was identified as Pseudomonas sp. based on the morphological, biochemical and physiological characteristics as well as 16S rRNA sequence analysis. In this study, an antimicrobial protein, named as CAP-1, was isolated from the culture of CMF-2 through ammonium sulfate precipitation and gel filtration chromatography. According to the result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), a major band indicated that the antimicrobial protein had a molecular mass of about 15 kDa, and it was identified as a hypothetical protein by MALDI-TOF-MS analysis and Mascot searching. CAP-1 displayed a broad antimicrobial spectrum against the indicator bacteria and fungus, including Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Candida albicans, especially some marine-derived microorganisms such as Vibrio vulnificus, Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio cholera, and Vibrio anguillarum, but showed little impact on tumor cells and normal human cells. The protein CAP-1 remained a stable antimicrobial activity in a wide range of temperature (20-80°C) and pH (2-10) conditions. These results suggested that CAP-1 might have a specific antimicrobial function not due to cytotoxicity.
从海月水母中分离出一株名为CMF-2的细菌菌株,其培养上清液表现出显著的抗菌活性。根据形态、生化和生理特征以及16S rRNA序列分析,菌株CMF-2被鉴定为假单胞菌属。在本研究中,通过硫酸铵沉淀和凝胶过滤色谱从CMF-2培养物中分离出一种抗菌蛋白,命名为CAP-1。根据十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结果,一条主要条带表明该抗菌蛋白的分子量约为15 kDa,通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析和Mascot搜索将其鉴定为一种假定蛋白。CAP-1对指示菌和真菌具有广泛的抗菌谱,包括金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌和白色念珠菌,特别是一些海洋来源的微生物,如创伤弧菌、溶藻弧菌、副溶血性弧菌、霍乱弧菌和鳗弧菌,但对肿瘤细胞和正常人细胞影响很小。蛋白CAP-1在广泛的温度(20-80°C)和pH(2-10)条件下保持稳定的抗菌活性。这些结果表明,CAP-1可能具有特定的抗菌功能,而非由于细胞毒性。