丙酮丁酸梭菌胞浆蛋白质组的系统研究

System Research of Cytoplasmic Proteome for Clostridium acetobutylicum

为了建立丙酮丁醇梭菌(Clostridium acetobutylicum)蛋白质组的研究方法,采用超声波破碎细胞,并结合丙酮沉淀分离蛋白的方法,提取丙酮丁醇梭菌胞浆蛋白质,发现加入罗氏蛋白酶抑制剂能有效地减少提取过程中蛋白质的降解。通过二维电泳技术对其胞浆蛋白进行有效分离,在pH 4～7的胶上分离出大约1230个蛋白点。利用基质辅助激光解析电离飞行时间质谱成功鉴定了19个蛋白质。这些蛋白质组研究方法对于缺少蛋白质组研究的梭菌属具有一定的参考价值。

To establish the proteomic methods of Clostridium acetobutylicum, the cytoplasmic proteins of C. acetobutylicum were extrcated by using sonication cell disruption method combined with acetone precipitation. The authors detected that added protease inhibitors cocktail （Roche diagnostics） significantly decreased protein degradation during sample preparation. The cytoplasmic proteins were separated by using two-dimensional polyacrylamide gel electrophesis, and approximately 1230 protein spots were detected on the pH 4-7 gel. 21 high abundance protein spots were excised and submitted to MALDI-TOF-MS/MS analysis. 19 protein spots were successfully identified by PMF and MS/MS. These proteomic methods will be useful for the Clostridium community of proteomic research.