The microbial enzymatic degradation of a painting from the Skokloster Palace was investigated. Lipolytic activity on the paint surface and cellulase activity in the canvas support were characterized. The isolated microorganisms cultured from samples taken from the painting were investigated for optimal growth conditions for both lipolytic and cellulase activities. Time of culture, pH, and humidity on synthetic media were varied. Penicillinase, EDTA, and high glucose concentrations were used to inhibit microbial growth and/or enzymatic activities.