Implementation of a Novel Method for Processing Proteins from Acetic Acid Bacteria via Liquid Chromatography Coupled with Tandem Mass Spectrometry
Juan J. Román-Camacho,
Juan C. Mauricio,
Irene Sánchez-León,
Inés M. Santos-Dueñas,
Carlos A. Fuentes-Almagro,
Francisco Amil-Ruiz,
Teresa García-Martínez,
Isidoro García-García
Affiliations
Juan J. Román-Camacho
Department of Agricultural Chemistry, Edaphology and Microbiology Agrifood Campus of International Excellence ceiA3, University of Cordoba, 14014 Cordoba, Spain
Juan C. Mauricio
Department of Agricultural Chemistry, Edaphology and Microbiology Agrifood Campus of International Excellence ceiA3, University of Cordoba, 14014 Cordoba, Spain
Irene Sánchez-León
Department of Agricultural Chemistry, Edaphology and Microbiology Agrifood Campus of International Excellence ceiA3, University of Cordoba, 14014 Cordoba, Spain
Inés M. Santos-Dueñas
Department of Inorganic Chemistry and Chemical Engineering, Agrifood Campus of International Excellence ceiA3, Institute of Chemistry for Energy and Environment (IQUEMA), University of Cordoba, 14014 Cordoba, Spain
Carlos A. Fuentes-Almagro
Proteomics Unit, Central Service for Research Support (SCAI), University of Cordoba, 14014 Cordoba, Spain
Francisco Amil-Ruiz
Bioinformatics Unit, Central Service for Research Support (SCAI), University of Cordoba, 14014 Cordoba, Spain
Teresa García-Martínez
Department of Agricultural Chemistry, Edaphology and Microbiology Agrifood Campus of International Excellence ceiA3, University of Cordoba, 14014 Cordoba, Spain
Isidoro García-García
Department of Inorganic Chemistry and Chemical Engineering, Agrifood Campus of International Excellence ceiA3, Institute of Chemistry for Energy and Environment (IQUEMA), University of Cordoba, 14014 Cordoba, Spain
Acetic acid bacteria (AAB) and other members of the complex microbiotas, whose activity is essential for vinegar production, display biodiversity and richness that is difficult to study in depth due to their highly selective culture conditions. In recent years, liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) has emerged as a powerful tool for rapidly identifying thousands of proteins present in microbial communities, offering broader precision and coverage. In this work, a novel method based on LC–MS/MS was established and developed from previous studies. This methodology was tested in three studies, enabling the characterization of three submerged acetification profiles using innovative raw materials (synthetic alcohol medium, fine wine, and craft beer) while working in a semicontinuous mode. The biodiversity of existing microorganisms was clarified, and both the predominant taxa (Komagataeibacter, Acetobacter, Gluconacetobacter, and Gluconobacter) and others never detected in these media (Asaia and Bombella, among others) were identified. The key functions and adaptive metabolic strategies were determined using comparative studies, mainly those related to cellular material biosynthesis, energy-associated pathways, and cellular detoxification processes. This study provides the groundwork for a highly reliable and reproducible method for the characterization of microbial profiles in the vinegar industry.
