Appl. Environ. Microbiol. 68: [2584][2585][2586][2587][2588] 2002). Here we demonstrate the aerobic bacterial catabolism of the ubiquitous toxic diaryl ether pollutant 1,2,3,4,7,8-hexachlorodibenzo-p-dioxin by this strain. The products of this biotransformation were identified as tetrachlorocatechol and 2-methoxy-3,4,5,6-tetrachlorophenol by comparing mass spectra recorded before and after nbutylboronate and N,O-bis(trimethylsilyl)-trifluoroacetamide derivatization with those of authentic compounds. Additional experiments showed that the less-chlorinated 1,2,3,7,8-pentachlorodibenzo-p-dioxin was not transformed by the strain RW1. The importance of substitution patterns for the degradability of individual congeners was illustrated by the fact that the 1,2,3-trichlorodibenzo-p-dioxin was catabolized to yield 3,4,5-trichlorocatechol, whereas the 2,3,7-trichlorodibenzo-p-dioxin was not attacked.Polychlorinated aromatic compounds such as the polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) are among the most problematic environmental pollutants because of their chemical inertness and recalcitrance, lipid solubility, and toxicity (1,3,8,13). Consequently, numerous studies have been carried out to elucidate the aerobic (7, 9, 19) bacterial degradation of PCDD/Fs, compounds produced by incineration processes and as unwanted by-products of the synthesis of pesticides and herbicides (4, 17). However, naturally occurring microorganisms have evolved to degrade and mineralize many, but by no means all, of these compounds. For example, several PCDD/Fs are degraded very slowly or not at all (12). In addition to genera such as Terrabacter (6,14) and Pseudomonas (6), several strains belonging to the genus Sphingomonas were reported to use diaryl ethers such as dibenzofuran and dibenzo-p-dioxin as the sole source of carbon and energy (3). Some of these isolates are examples for effective biocatalysts that can even catabolize some di-, tri-, and tetrachlorinated dibenzo-p-dioxins and dibenzofurans (6,7,9,19). However, the aerobic catabolism of important environmental pollutants such as 1,2,3,7,8-penta-or 1,2,3,4,7,8-hexachlorodibenzo-p-dioxin, giving rise to the formation of corresponding metabolites, has not been demonstrated previously.The catabolism of dibenzofurans and dibenzo-p-dioxins is in Sphingomonas wittichii RW1 initiated by a ring-hydroxylating dioxygenase (2,18,20), introducing two hydroxyl groups in the accessible angular positions of the diaryl ether, thereby producing unstable hemiacetals. The labile intermediate spontaneously decays in the case of dibenzo-p-dioxins to yield the corresponding 2,2Ј,3-trihydroxydiphenyl ether, which is further processed, yielding a catechol and the corresponding 2-hydroxy-cis,cis-muconic acid (3, 21). However, despite the fact that previous transformation experiments monitoring the disappearance of selected mono-to tetrachlorinated congeners of dibenzofurans and dibenzo-p-dioxins have indicated some depletion of tetrachlorodibenzo-p-dioxins is due to bact...