基于核糖体蛋白质标志物及MALDI-TOF MS技术快速鉴定伦茨菌属放线菌

Rapid identification of Lentzea members:based on ribosomal protein biomarkers and MALDI-TOF MS

【目的】伦茨菌属(Lentzea)放线菌(Actinobacteria)代谢产物具有广泛的生物活性,在医药领域展现出潜在的应用价值。本研究尝试建立以核糖体蛋白质为标志物,利用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization-time of flight mass spectrometry,MALDI-TOF MS)技术鉴定伦茨菌属放线菌的方法。【方法】检索基因组数据库,提取伦茨菌属菌种模式菌株15种核糖体蛋白质的序列并计算理论分子量;通过分子量比对分析伦茨菌属菌种模式菌株之间及其与邻近属菌种模式菌株之间15种核糖体蛋白质的匹配度,提出鉴定至菌种及属的核糖体蛋白质匹配数标准;选取目标属和非目标属菌种进行MALDI-TOF MS测试和分析并修正鉴定标准。【结果】将待测菌株的MALDI-TOF质谱峰与伦茨菌属各菌种模式菌株的15种核糖体蛋白质分别匹配,通过最大匹配数及质谱峰强度模式可鉴定至属或种。【结论】本研究建立了基于15种核糖体蛋白质标志物及MALDI-TOF MS技术鉴定伦茨菌属放线菌的方法,可为放线菌纲其他类群的快速鉴定提供借鉴。

[Objective]Metabolites from members of Lentzea(Actinobacteria)have a wide range of bioactivities,thus showing potential application value in biopharmaceutical field.This study aims to establish a method to identify members of Lentzea based on ribosomal protein biomarkers and matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS).[Methods]We searched public genome database to obtain the sequences of 15 ribosomal proteins from the type strains of Lentzea and calculated the theoretical molecular weights.The molecular weight comparison was performed to investigate the matching of 15 ribosomal proteins between the type strains of Lentzea and that between the type strains of Lentzea and its closely related genera(Actinosynnema and Saccharothrix).Identification criteria for Lentzea at the species and genus level were proposed based on the number of ribosomal protein matches.Subsequently,the identification criteria were tested with target and non-target organisms and improved.[Results]MALDI-TOF MS peaks from an unknown organism were compared with the weight list of 15 ribosomal protein biomarkers from each Lentzea type strain and the number of peaks that matched each strain’s biomarkers was obtained.Based on the maximum of matches and peak intensity mode,the organism can be identified at the genus or species level.[Conclusion]This study established a method for identifying Lentzea members based on 15 ribosomal protein biomarkers and MALDI-TOF MS,which can provide a reference for the rapid identification of other Actinobacteria groups.