Aims Pulsed light (PL) technology is a surface decontamination process that can be used on food, packaging or water. PL efficiency may be limited by its low degree of penetration or because of a shadow effect. In these cases, surviving bacteria will be able to perceive PL as a stress. Such a stress was mimicked using low transmitted energy conditions, and its effects were investigated on the highly environmental adaptable bacterium Enterococcus faecalis V583. Methods and Results In these laboratory conditions, a complete decontamination of the artificially inoculated medium was performed using energy doses as low as 1.8 J cm-2, while a treatment of 0.5, 1 and 1.2 J cm-2 led to a 2.2, 6 and 7-log10 CFU ml-1 reduction in the initial bacterial population, respectively. Application of a 0.5 J cm-2 pretreatment allowed the bacteria to resist more efficiently a 1.2 J cm-2 subsequent PL dose. This 0.5 J cm-2 treatment increased the bacterial mutation frequency and affected the abundance of 19 proteins as revealed by a global proteome analysis. Conclusions Enterococcus faecalis is able to adapt to a PL treatment, providing a molecular response to low-energy PL dose, leading to enhanced resistance to a subsequent treatment and increasing the mutation frequency. Significance and Impact of the Study This study gives further insights on Ent. faecalis capacities to adapt and to resist to stress.