It was confirmed that malodor connected with an air-conditioner in an automobile is caused by microbial volatile organic compounds (MVOCs) produced by microorganisms and through microorganisms coexisting with each other to form a biofilm on the evaporator surface. A bacterium, Methylobacterium aquaticum, can form a biofilm on the evaporator surface. The biofilm was composed of 45.79% C (Carbon), 42.36% O (Oxygen), 1.85% Na (Sodium), 5.42% Al (Aluminum), 1.39% P (Phosphorus), 0.74% Cl (Chlorine) and 2.45% K (Potassium). This result matches the composition of the biofilm formed on the surface of the used evaporator. It was determined that sulfur compounds (Hydrogen sulfide, Dimethyl sulfide) and organic acids (n-Butyric acid, n-Valeric acid, iso-Valeric acid) in the air which was blown into the automobile were generated by Methylobacterium aquaticum and Aspergillus versicolor, respectively. On the other hand, volatile organic compounds (Toluene, Xylene, 2-Ethylhexanol, 2-Phenyl- 2-propanol, Ethylbenzene) were not found. It is estimated that the reason is due to the low concentration of generated MVOCs or is caused by the change of some MVOCs depending on the nutrients (medium).