Degradation of unsaturated hydrocarbons by methanogenic enrichment cultures

The biodegradability of hydrocarbons under anaerobic conditions was studied in enrichment cultures using mineral media inoculated with sewage sludge or sediment samples of limnic and marine origin. No indication of methanogenic degradation was obtained with either n-hexane, nhexadecane, n-heptadecane, 1-hexene, cis-2hexene, trans-2-hexene, isoprene, 1-hexine, benzene, toluene, xylene, cyclohexene, cycloheptatriene, cyclopentadiene, styrene, naphthalene, azulene, or fl-carotene. Squalene was incompletely converted to methane and carbon dioxide. Complete degradation was observed with 1-hexadecene. Methanogenic subcultures were maintained on 1hexadecene and squalene. Both enrichments contained after several transfers Methanospirillum hungatei and Methanothrix soehngenii as prevalent methanogenic bacteria. Acetate (~< 80 #M) was the only intermediary product detected indicating that degradation proceeded via hydrogen-dependent syntrophic fl-oxidations. Short rods on hexadecene and cocci on squalene were found to be associated with substrate degradation. The resuits indicate that terminal double bonds can be sufficient to allow methanogenic degradation of hydrocarbons whereas branching and terminal ring closures may significantly contribute to hydrocarbon stability in anoxic environments.