MALDI-TOF MS analysis of ribosomal proteins coded in S10 and spc operons rapidly classified the Sphingomonadaceae as alkylphenol polyethoylate-degrading bacteria from the environment
MALDI-TOF MS analysis of ribosomal proteins coded in S10 and spc operons rapidly classified the Sphingomonadaceae as alkylphenol polyethoylate-degrading bacteria from the environment
复制标题
对 S10 和 spc 操纵子编码的核糖体蛋白进行 MALDI-TOF MS 分析，将鞘氨醇单胞菌科快速分类为环境中的烷基酚聚乙氧基化物降解细菌
DOI：
10.1111/j.1574-6968.2012.02525.x
复制
发表时间：
2012
影响因子：
2.1
通讯作者：
Yudai Hotta
中科院分区：
文献类型：
--
作者：
Chihiro Tachinami;Hiroki Senshu;Shigeru Ida;藤原幹;Yudai Hotta
采用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）技术，以S10-spc-alpha操纵子中编码的核糖体亚基蛋白为生物标记物，对环境中的鞘氨醇单胞菌进行分类。为构建核糖体蛋白质数据库，根据基因组测序菌株的核苷酸序列设计特异性引物，对鞘氨醇单胞菌科（Sphingomonadaceae）模式菌株及其相关烷基酚聚氧乙烯醚（APEOn）降解菌的S10-spc-α操纵子序列进行测序。观察到的完整细胞的MALDI质谱与构建的核糖体蛋白质数据库的理论质量进行了比较。S10-spc-α操纵子编码的9个生物标志物L18、L22、L24、L29、L30、S 08、S14、S17和S19可以成功区分Sphingopyxis terraeNBRC 15098 T和APEOn降解菌BSN 20，尽管16 S rRNA基因序列只有一个碱基差异。S10-GERMS（S10-spc-alphaoperon gene-encoded ribosomal protein mass spectrum）方法是一种在菌株水平上区分鞘氨醇单胞菌的有效方法，可用于环境中主要的APEOn降解菌的检测和监测。
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) using ribosomal subunit proteins coded in theS10–spc-alphaoperon as biomarkers was applied for the classification of theSphingomonadaceaefrom the environment. To construct a ribosomal protein database,S10-spc-alphaoperon of type strains of theSphingomonadaceaeand their related alkylphenol polyethoxylate (APEOn)-degrading bacteria were sequenced using specific primers designed based on nucleotide sequences of genome-sequenced strains. The observed MALDI mass spectra of intact cells were compared with the theoretical mass of the constructed ribosomal protein database. The nine selected biomarkers coded in theS10-spc-alphaoperon, L18, L22, L24, L29, L30, S08, S14, S17, and S19, could successfully distinguish theSphingopyxis terraeNBRC 15098Tand APEOn-degrading bacteria strain BSN20, despite only one base difference in the 16S rRNA gene sequence. This method, named theS10-GERMS (S10-spc-alphaoperon gene-encoded ribosomal protein mass spectrum) method, is a significantly useful tool for bacterial discrimination of theSphingomonadaceaeat the strain level and can detect and monitor the main APEOn-degrading bacteria in the environment.