Protein profiling analysis based on matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry and its application in typing Streptomyces isolates

• Published in: Talanta (Oxford) 2020-02, Vol.208, p.120439-120439, Article 120439
• Main Authors: Sun, Xiaoshan, Wu, Peichun, Zhao, Chunxia, Zheng, Fujian, Hu, Chunxiu, Lu, Xin, Xu, Guowang
• Format: Article
• Language: English
• Subjects: Activity screening; Cluster analysis; Matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry; Protein profiling; Streptomyces isolates
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2019.120439

Marine Streptomyces is a potential source of novel bioactive natural products in medicine and agriculture. The current discrimination and screening method of Streptomyces isolates is not accurate and time-consuming, and a novel method is necessary. In this study, a protein profiling method based on an ultrahigh resolution 15 T Matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS) was established and applied for differentiation and bioactivity screening of marine Streptomyces isolates. To obtain robust protein profiling, the effects of the protein extraction method, the matrix-solvent, the sample deposition mode, and the culture time of isolates on protein profiling were thoroughly studied, the optimal conditions were obtained. To evaluate the performance of the developed MALDI-FTICR MS method, MALDI-time of flight (TOF) MS and 16S rRNA were applied in parallel to analyze 25 marine Streptomyces isolates. We found that the clustering result of MALDI-FTICR MS was more similar to that of 16S rRNA than MALDI-TOF MS. And MALDI-FTICR MS could effectively indicate the antibacterial activity of Streptomyces isolates against three plant pathogenic bacteria including Xanthomonas campestris, Xanthomonas oryzae and Erwinia carotovora. Furthermore, a differential protein/peptide was defined and successfully applied to predict antibacterial activity of blind samples. This study demonstrated that MALDI-FTICR MS has great potential to discriminate and screen complex microorganisms, especially those closely related strains. [Display omitted] •A robust protein profiling method by a 15T MALDI-FTICR MS was developed.•The method showed good performance to differentiate Streptomyces isolates.•Isolates with antibacterial activity were effectively identified.•A specific protein/peptide related to antibacterial activity was obtained.