Biological contamination of surfaces, both in industry and in health care, plays an important role as a potential vector of disease transmission. Metals have been described to be effective antibiofilm agents, and the efficacy of silver ions as a disinfectant has been known for centuries. The activity of AgNO3 combined with the lipopeptide biosurfactant V9T14 has been studied against a preformed Escherichia coli biofilm on the Calgary Biofilm Device. Results indicated that the activity of silver can be synergistically enhanced by the presence of V9T14, both allowing for a reduction in the quantity of silver used and for greater antimicrobial activity. The concentration of silver needed to obtain this reduction in the silver-biosurfactant solution was from 129- to 258-fold less than the concentration of silver alone. To our knowledge, this is the first time that a synergistic interaction between a lipopeptide biosurfactant and silver has been observed.