Instituto de Química Orgánica General (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain.
Anal Chim Acta. 2010 Jan 18;658(1):81-6. doi: 10.1016/j.aca.2009.10.053. Epub 2009 Oct 29.
Capillary electrophoresis (CE) coupled with laser-induced fluorescence detection (LIF) has allowed to obtain protein fingerprints, which have demonstrated to be useful in microorganisms characterization. In this work, protein fingerprints of two species of Staphylococcus grown in different culture media and submitted to temperature and nitrosative stress were studied by CE-LIF. After the growth of the bacteria, protein extracts were obtained by cell lysis using sonication. The water-soluble fraction of these lysates was derivatized on-capillary with a fluorogenic dye, 3-(2-furoyl)quinoline-2-carboxaldehyde. The fluorescent products were analyzed by CE using phosphate buffer containing submicellar concentrations of sodium pentanesulfate and detected by LIF. Different protein fingerprints were obtained depending on the bacterial species studied, indicating the usefulness of this method for the identification of different species of the same bacterial genus. It was also demonstrated that the CE protein fingerprints were dependent on the culture conditions, such as growth medium, or on stressing conditions, such as heat shock or nitrosative stress.
毛细管电泳(CE)与激光诱导荧光检测(LIF)相结合,能够获得蛋白质指纹图谱,这已被证明在微生物特征描述中非常有用。在这项工作中,通过 CE-LIF 研究了在不同培养介质中生长并经受温度和硝化应激的两种葡萄球菌的蛋白质指纹图谱。细菌生长后,通过超声细胞裂解获得蛋白质提取物。这些裂解物的水溶性部分在毛细管内用荧光染料 3-(2-呋喃酰基)喹啉-2-甲醛衍生化。使用含有亚胶束浓度的戊烷磺酸钠的磷酸盐缓冲液通过 CE 分析荧光产物,并通过 LIF 检测。根据所研究的细菌种类,得到了不同的蛋白质指纹图谱,表明该方法可用于鉴定同一细菌属的不同物种。还证明了 CE 蛋白质指纹图谱取决于培养条件,例如生长培养基,或应激条件,例如热休克或硝化应激。