恶臭假单胞菌好氧降解高氯联苯的蛋白质组分析

Proteomic analysis of aerobic degradation of highly chlorinated PCBs by Pseudomonas putida HP1

利用固相pH梯度双向凝胶电泳分离恶臭假单胞菌以葡萄糖和Aroclor1260作为营养基质生长的菌株总蛋白,凝胶经银蓝显色后,采用PDQuest图像分析软件比较、分析识别差异表达蛋白,应用MOLDI-TOF-MS得到相应的肽质量指纹图谱,然后搜索数据库鉴定部分差异蛋白点.结果表明,获得了分辨率较高、重复性较好的不同营养基质条件下菌株蛋白的双向凝胶电泳图谱,比较分析共发现80个差异蛋白点,成功鉴定14个差异表达蛋白,包括应激应答蛋白、蛋白质生物合成、运载体和代谢相关酶类.提示与葡萄糖作为生长基质相比,菌体以多氯联苯作为碳源和能源生长时,产生应激,通过增强应激响应蛋白、蛋白质生物合成相关蛋白和相关代谢酶类的表达使细胞在胁迫条件下维持自身的稳定性与生长代谢.

The total proteins of Pseudomonas putida HP1 that use glucose and Arcolor 1260 as nutritional matrix for growth were separated respectively by immobilized pH gradient-based 2-DE.The differential expressed proteins were screened by PD Quest analysis software and identified by peptide mass fingerprint based on MOLDI-TOF-MS.Part of differential protein spots were identified in reference of the database.These results showed that a well-resolved and reproducible 2-DE pattern was obtained,and eighty differential expressed protein spots were found by comparative analysis.Fourteen differential expressed proteins were identified,including stress response proteins,proteins related to protein biosynthesis,transporter and metabolic enzymes.This suggested that the cell using PCBs as sole carbon and energy source produced stress response,and then maintained their stability,growth and metabolism by enhancing the expression of stress response proteins,proteins related to protein biosynthesis and enzymes related to metabolism.