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Novel organization of genes in a phthalate degradation operon of Mycobacterium vanbaalenii PYR-1

¹ Division of Microbiology, National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA
² Institute of Molecular Biology, Slovak Academy of Sciences, 845 51 Bratislava, Slovakia

Correspondence
Ashraf A. Khan
Ashraf{at}nctr.fda.gov

Mycobacterium vanbaalenii PYR-1 is capable of degrading polycyclic aromatic hydrocarbons (PAHs) to ring cleavage metabolites. This study identified and characterized a putative phthalate degradation operon in the M. vanbaalenii PYR-1 genome. A putative regulatory protein (phtR) was encoded divergently with five tandem genes: phthalate dioxygenase large subunit (phtAa), small subunit (phtAb), phthalate dihydrodiol dehydrogenase (phtB), phthalate dioxygenase ferredoxin subunit (phtAc) and phthalate dioxygenase ferredoxin reductase (phtAd). A 6·7 kb EcoRI fragment containing these genes was cloned into Escherichia coli and converted phthalate to 3,4-dihydroxyphthalate. Homologues to the operon region were detected in a number of PAH-degrading Mycobacterium spp. isolated from various geographical locations. The operon differs from those of other Gram-positive bacteria in both the placement and orientation of the regulatory gene. In addition, the M. vanbaalenii PYR-1 pht operon contains no decarboxylase gene and none was identified within a 37 kb region containing the operon. This study is the first report of a phthalate degradation operon in Mycobacterium spp.

The GenBank/EMBL/DDBJ accession numbers for the sequences reported in this paper are AY365117 (M. vanbaalenii PYR-1 pht operon region), AY372761 and AY372763 (Mycobacterium sp. PAH2.135 phtAa and phtB PCR products, respectively), and AY372762 and AY372764 (M. flavescens PYR-GCK phtAa and phtB PCR products, respectively).

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