Seven gram-negative, aerobic bacteria were isolated from a mixed culture enriched for asphalt-degrading bacteria. The predominant genera of these isolates were Pseudomonas, Acinetobacter, Alcaligenes, Flavimonas, and Flavobacterium. The mixed culture preferentially degraded the saturate and naphthene aromatic fractions of asphalt cement-20. A residue remained on the surface which was resistant to biodegradation and protected the underlying asphalt from biodegradation. The most potent asphalt-degrading bacterium, Acinetobacter calcoaceticus NAV2, excretes an emulsifier which is capable of emulsifying the saturate and naphthene aromatic fractions of asphalt cement-20. This emulsifier is not denatured by phenol.