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Peptide biomarkers for biogenic amine-producing bacterial characterization by LC-ESI-MS/MS

AuthorsAbril, A. G.; Ortea, Ignacio ; Osorio, A. P.; Rama, J. L. R.; Carrera, Mónica ; Böhme, K.; Barros-Velázquez, J. ; Cañas, Benito; Villa, T. G.; Calo-Mata, Pilar
Issue Date2019
CitationCongreso Nacional de Biotecnología Biotec (2019)
AbstractSome bacterial strains have the ability to decarboxylate specific amino acids producing biogenic amines (such as histamine from histidine). The detection of these strains having amino acid decarboxylase activity is important to lower the risk of biogenic amine in food products, especially histamine. Histamine is regulated in fish (lower than 100 mg/Kg-1, but it is generally recognized that the presence of other biogenic amines aggravate symptoms from histamine. Molecular methods for the detection of biogenic amine-producing bacteria in foods are based on the specific detection of decarboxylase genes by PCR allowing the detection before the biogenic amine is produced [1]. However, the difficulty is the availability of universal primers. Mass spectrometry [2] can provide a useful tool for the detection of strain-specific diagnostic decarboxylase peptides. In this study, proteins from biogenic amine producing strains were isolated, after tryptic digestion, the resulting peptides were analysed by LC-ESI-MS/MS (Liquid chromatography-electrospray ionizationtandem mass spectrometry). A total of 100μg protein extraction was digested with trypsin, cleaned on a C18 microSpinTM, following by LC-MS/MS analyse. Proteomic data was processed by SEQUEST (Proteome Discoverer 1.4 package, Thermo Scientific) against Bacteria in the UniProt/ TrEMBL database. MS data was analysed to identify the peptides from the amino acid decarboxylases and then compared to the GenBank database using the Blast tool (NCBI) to achieve peptide/protein identification. Thus, decarboxylase peptides were detected in studied strains that could be used as biomarkers in further studies
DescriptionPoster.-- Congreso Nacional de Biotecnología Biotec, Vigo, 10-13 de junio de 2019
Appears in Collections:(IIM) Comunicaciones congresos
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