Our study aimed to investigate the proteome structure of R. akari and unveil immunoreactive surface-exposed proteins using the latest proteomics techniques. To achieve these goals, we applied the Shotgun protein identification approach; membrane surface protein enrichment methods combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and immunoblotting techniques. We identified aroun 300 unique proteins from the whole-cell extract exhibiting that the most represented proteins were those involved in translation, post-translational modifications, energy production, and cell wall development. A significant part belongs to amino acid transport and intracellular trafficking. Besides, several proteins affecting virulence were detected. Then, in silico analysis from detected surface proteins revealed that 25 putative predicted outer membrane proteins contain beta-barrel structure and 11 proteins were predicted to have secretion signal peptide sequence.