Analysis of bacterial proteomes can be used to obtain large amounts of information about adaptive microbial mechanisms to changing extracellular conditions. In the past, many bacterial species with the ability to degrade acrylamide were isolated. In this study differences in the Enterococcus faecalis proteome upon acrylamide exposure were investigated. We revealed substantial changes in the proteome of bacteria cultured in different environmental conditions. Microorganisms exposed to acrylamide showed higher accumulation of proteins associated with energy metabolism and its regulation. Moreover, several proteins involved in protection of cells from stress conditions were also identified. These biomacromolecules are involved in proper folding of newly synthesized polypeptides like chaperones or participate in mechanisms of DNA repair. In contradiction with previous reports, the presence of amidase was not detected. However, identification of aminopeptidase with activity to hydrolyze amino acid amides can indicate that it can degrade acrylamide to acrylic acid and ammonia instead of amidase. According to identified proteomic profiles, a new mechanism of acrylamide degradation by Enterococcus faecalis is proposed.