短小芽孢杆菌胞外蛋白质组与蛋白酶的研究

项目名称： 短小芽孢杆菌胞外蛋白质组与蛋白酶的研究

项目编号： No.31070071

项目类型： 面上项目

立项/批准年度： 2011

项目学科： 生物科学

项目作者： 王海燕

作者单位： 四川大学

项目金额： 8万元

中文摘要： 本实验室早期筛选到一株能产生碱性蛋白酶的短小芽孢杆菌，其发酵液具有良好的生皮脱毛效果。本课题以该短小芽孢杆菌菌株SCU11为材料，通过液相色谱与质谱联用，进行了胞外蛋白质全谱分析。研究了碱性胁迫下的短小芽孢杆菌胞外分泌蛋白的差异，通过双向电泳及质谱分析，鉴定出发酵液中的10种差异蛋白，对这些蛋白的细胞定位及参与的代谢过程进行了生物信息学分析。从短小芽孢杆菌SCU11中克隆到vpr、wpr、aprX 、bpf 4个胞外蛋白酶基因，它们与短小芽孢杆菌SAFR-032菌株相应基因的氨基酸序列同源性分别为：96%、90%、94%及70%，显示两个菌株之间存在较大差异。利用比较基因组学方法预测了短小芽孢杆菌转录调控网络，对短小芽孢杆菌胞外蛋白酶基因的候选转录因子进行了预测。

中文关键词： 短小芽孢杆菌；蛋白全谱分析；碱胁迫；蛋白酶；双向电泳

英文摘要： A novel B. pumilus strain SCU11 was screened out for its ability to produces alkaline protease with excellent dehairing effect in our former researches. In this project, the extracellular proteome of B. pumilus SCU11 was resolved by a combination of HPLC and LC-MS/MS. The changes of the extracellular proteome of B pumilus SCU11 in response to alkali stress was studied by 2-DE. Ten protein spots significantly affected by alkaline stress were identified by MALDI-TOF-TOF. All of the identified proteins were analyzed by SignalP to characterize their predicted mode of secretion. Four protease genes, including vpr,wpr,aprX and bpf, were cloned from SCU11 and sequenced. A transcriptional regulatory network of B pumilus was constructed by comparative genomics methods and some candidate transcription factors for protease were predicted.

英文关键词： Bacillus pumilus; extracellular proteome; alkali stress; protease; 2-DE