【目的】糖丝菌属(Saccharothrix)是一类丝状稀有放线菌，在生物医药、工业酶制剂和环境修复等领域展现出应用价值。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)技术鉴定糖丝菌属放线菌的方法。【方法】检索基因组数据库，提取糖丝菌属测序菌株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糖丝菌属不同菌种之间及其模式菌株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质谱峰与糖丝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、质谱峰强度模式及特征质谱峰鉴定至属或种。【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技术鉴定糖丝菌属放线菌的方法，可用于定向筛选和快速鉴定糖丝菌。
[Objective] Saccharothrix is a genus of rare filamentous actinomycetes showing great application values in the biopharmaceutical field, industrial enzyme preparations, and environmental remediation. This study aims to establish a method to identify Saccharothrix spp. based on ribosomal protein biomarkers and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). [Methods] We searched two public genome databases to obtain the sequences of 15 ribosomal proteins from the genome-sequenced strains of Saccharothrix and calculated the theoretical molecular weights. We then compared the molecular weights to investigate the matching of 15 ribosomal proteins between different species of Saccharothrix and that between the type strains of Saccharothrix and its closely related three genera (Umezawaea, Actinosynnema, and Lentzea). The criteria for identifying Saccharothrix at the species and genus levels were proposed based on the number of ribosomal protein matches. Subsequently, the identification criteria were tested by MALDI-TOF MS 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 Saccharothrix type strain and the number of peaks that matched each strain’s biomarkers was obtained. Based on the maximum matches, peak intensity mode, and characteristic peaks, the organism can be identified at the genus or species level. [Conclusion] This study established a method for identifying Saccharothrix spp. based on 15 ribosomal protein biomarkers and MALDI-TOF MS, which can be used in directed screening and rapid identification of Saccharothrix spp.
孙伟,许杰,张薇,罗倩,台萃. 基于核糖体蛋白质标志物及基质辅助激光解吸电离飞行时间质谱技术快速鉴定糖丝菌属放线菌. 微生物学报, 2023, 63(12): 4800-4813复制