The characteristics of de-emulsification of pertroleum emulsion by Nocardia amarae were investigated. Insoluble medium containing n-hexadecane was more effective than soluble medium in de-emulsification of emulsion containing diesel and bunker C as the organic phase. Emulsion made by the addition of xanthan or bioemulsifier was de-emulsified by N. amarae, and longer culture age was effective. In low viscosity range, organic phase with longer carbon chain was more effective. The contact, angle between bacterial film and water droplet in air increased from 16 degree for 4 day culture age to 26 degree for 15 day. The contact angle between bacterial film and water droplet in kerosene, n-heyxane or n-hexadecane also increased to greater than 100 degree after 3 day culture age. The hydrophobicity of bactgerial film increased according to the culture age.