Staphylococcus sp. strain PN/Y, capable of utilizing phenanthrene as a sole source of carbon and energy, was isolated from petroleum-contaminated soil. In the degradation of phenanthrene by strain PN/Y, various metabolites, isolated and identified by a combination of chromatographic and spectrometric analyses, revealed a novel phenanthrene assimilation pathway involving 2-hydroxy-1-naphthoic acid. Metabolism of phenanthrene was initiated by the dioxygenation on the 1,2-position of phenanthrene followed by meta-cleavage of phenanthrene-1,2-diol, leading to 2-hydroxy-1-naphthoic acid, which was then processed via a novel meta-cleavage pathway, leading to the formation of trans-2,3-dioxo-5-(29-hydroxyphenyl)-pent-4-enoic acid and subsequently to salicylic acid. In the lower pathway, salicylic acid was transformed to catechol, which was then metabolized by catechol-2,3-dioxygenase to 2-hydroxymuconaldehyde acid, ultimately forming TCA cycle intermediates. The catabolic genes involved in phenanthrene degradation were found to be plasmid-encoded. This detailed study of polycyclic aromatic hydrocarbon (PAH) metabolism by a Gram-positive species involving a unique ring-cleavage dioxygenase in a novel phenanthrene degradation pathway provides a new insight into the microbial degradation of PAHs.
INTRODUCTIONPolycyclic aromatic hydrocarbons (PAHs) constitute a group of priority environmental pollutants, which are ubiquitous contaminants in soils and sediments and are of environmental concern because of their toxic, mutagenic and/or carcinogenic effects (Mastrangelo et al., 1996;Marston et al., 2001;Xue & Warshawsky, 2005). In recent years, the biodegradation of PAHs has received considerable attention and a variety of micro-organisms have been reported to play important roles in the process (Pothuluri & Cerniglia, 1994;Shuttleworth & Cerniglia, 1995;Kanaly & Harayama, 2000;Habe & Omori, 2003;Tortella et al., 2005). Bioremediation technologies have increasingly been proposed to decontaminate PAH-contaminated sites (Harayama, 1997;Samanta et al., 2002;Parrish et al., 2004;Vinas et al., 2005).Phenanthrene, a PAH with three condensed rings fused in angular fashion, has a 'bay-region' and a 'K-region' and is often used as a model substrate for studies on the metabolism of carcinogenic PAHs. Over the last 60 years, a number of studies on phenanthrene degradation by several Gram-negative and Gram-positive bacterial species have been reported (Evans et al., 1965;Kiyohara et al., 1976Kiyohara et al., , 1982Kiyohara & Nagao, 1978;Barnsley, 1983;Gibson & Subramanian, 1984;Houghton & Shanley, 1994;Adachi et al., 1999;Samanta et al., 1999), where various pathways and metabolic diversity involved in phenanthrene degradation were documented. In general, the metabolic pathway is initiated by the double hydroxylation of the bay-region of phenanthrene by a dioxygenase enzyme to form cis-3,4-phenanthrenedihydrodiol. The resultant dihydrodiol is then converted by the action of dihydrodiol dehydrogenase to 3,4-dihydroxyphenanthrene, which und...