中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 乳杆菌胞外蛋白提取鉴定与双向电泳图谱建立 农业机械学报 2016,47 (9) 234-240
Position: Home > Articles > Extraction and Identification of Extracellular Proteins from Lactobacilli and a Reference Extracellular Proteomic Map of Lactobacillus paracasei L14 Transactions of the Chinese Society for Agricultural Machinery 2016,47 (9) 234-240

乳杆菌胞外蛋白提取鉴定与双向电泳图谱建立

作  者：
赵伟;郭慧媛;任发政;陈尚武
单  位：
中国农业大学食品科学与营养工程学院
关键词：
乳杆菌;胞外蛋白;提取方法;蛋白质组;双向电泳图谱
摘  要：
采用三氯乙酸(TCA)-丙酮沉淀法提取乳杆菌胞外蛋白并进行了优化,通过SDS-PAGE分析了该方法用于7株乳杆菌胞外蛋白的提取效果,同时对乳杆菌不同生长期胞外蛋白的分泌特性进行了研究,采用MALDI-TOF/TOFMS鉴定了胞外蛋白,最后通过双向电泳(2-DE)分析了L.paracasei L14的胞外蛋白组。结果表明:采用终质量分数为6%的TCA提取乳杆菌胞外蛋白效果最好,并且具有通用性;从乳杆菌稳定期提取出种类较多的胞外蛋白,成功鉴定出L.paracasei SW2胞外蛋白中3个蛋白质;使用半合成培养基培养乳杆菌,可以提取出符合2-DE样品纯度要求的胞外蛋白,从L.paracasei L14胞外蛋白组2-DE图谱上检测到(130±10)个蛋白质点,约占L.paracasei预测分泌蛋白组的46%。
译  名：
Extraction and Identification of Extracellular Proteins from Lactobacilli and a Reference Extracellular Proteomic Map of Lactobacillus paracasei L14
作  者：
Zhao Wei;Guo Huiyuan;Ren Fazheng;Chen Shangwu;College of Food Science and Nutritional Engineering,China Agricultural University;
关键词：
Lactobacillus;;extracellular proteins;;extraction method;;proteome;;2-D electrophoresis
摘  要：
Lactobacillus extracellular proteins are key molecular factors in promoting the probiotic-host crosstalk and signal transduction. Their extractions and identifications could contribute to unravelling the probiotic molecular mechanisms such as adhesion to intestinal surfaces,modulating immune response and enhancing antagonism towards pathogens. A protocol based on trichloroacetic acid( TCA)-acetone precipitation of proteins was presented and optimized. The method was tested on seven Lactobacillus strains after sodium dodecyl sulfate-polyacrylamide gel electrophoresis( SDS-PAGE). The extracellular proteins secretion capacity of L. paracasei SW2 in different growth phases was studied and some protein bands were selected for identification by tandem MS. The extracellular proteome of L. paracasei L14 were analyzed by 2-D electrophoresis( 2-DE). The results showed that the final concentration of 6% TCA was the most suitable for extracting extracellular proteins from lactobacilli. More extracellular proteins could be extracted in the stationary phase than mid-exponential growth phase. Three of extracellular proteins extracted from L. paracasei SW2 were successfully identified. The extracellular samples extracted from semi-defined medium( SDM) could be analyzed by 2-DE. 130 ± 10 spots,corresponding to about 46%coverage of the predicted secretome of L. paracasei,were detected in the 2-DE map of L. paracasei L14 extracellular proteome. In conclusion,this study preliminarily established the extraction method and 2-DE analysis system of extracellular proteins from lactobacilli and paved the way for a more comprehensive insight into relevant research.

相似文章

导出引用
计量
文章访问数: 11
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46