Tuberculosis (TB) is one of major causes of death worldwide. Bacillus Calmette-Guerin (BCG) is the only licensed TB vaccine and its inability to protect against adult pulmonary TB can be due to genetic differences among strains described since the 1940s. In this work, we compared the proteomic profile of the surface-associated proteins from M. bovis BCG Moreau, the Brazilian vaccine strain, and the BCG Pasteur reference strain. The methodology used was 2D-gel electrophoresis combined with mass spectrometry techniques (MALDI-TOF/TOF). We identified 116 proteins and 19 were more abundant in BCG Moreau whereas in Pasteur only 8 proteins presented higher levels of amount. Bioinformatic analysis using the programs MoonProt and MultiTask could reveal 27 moonlighting function among the 116 identified proteins