Characteristics of phenanthrene-degrading bacteria isolated from soils contaminated with polycyclic aromatic hydrocarbons

Ten bacterial strains were isolated from seven contaminated soils by enrichment with phenanthrene as the sole carbon source. These isolates and another phenanthrene-degrading strain were examined for various characteristics related to phenanthrene degradation and their ability to metabolize 12 other polycyclic aromatic hydrocarbons (PAH), ranging in size from two to five rings, after growth in the presence of phenanthrene. Fatty acid methyl ester analysis indicated that at least five genera (Agrobacterium, Bacillus, Burkholderin, Pseudomonas, and Sphingomonas) and at least three species of Pseudomonas were represented in this collection. All of the strains oxidized phenanthrene according to Michaelis-Menten kinetics, with half-saturation coefficients well below the aqueous solubility of phenanthrene in all cases. All but one of the strains oxidized 1-hydroxy-2-naphthoate following growth on phenanthrene, and all oxidized at least one downstream intermediate from either or both of the known phenanthrene degradation pathways. All of the isolates could metabolize (oxidize, mineralize, or remove from solution) a broad range of PAH, although the exact range and extent of metabolism for a given substrate were unique to the particular isolate. Benz[a]anthracene, chrysene, and benzo[a]pyrene were each mineralized by eight of the strains, while pyrene was not mineralized by any. Pyrene was, however, removed from solution by all of the isolates, and the presence of at least one significant metabolite from pyrene was observed by radiochromatography for the five strains in which such metabolites were sought. Our results support earlier indications that the mineralization of pyrene by bacteria may require unique metabolic capabilities that do not appear to overlap with the determinants for mineralization of phenanthrene or other high molecular weight PAH.