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Microbiology 152 (2006), 1929-1940; DOI  10.1099/mic.0.28783-0
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Microbiology 152 (2006), 1929-1940; DOI  10.1099/mic.0.28783-0
© 2006 Society for General Microbiology

Identification and functional analysis of the genes for naphthalenesulfonate catabolism by Sphingomonas xenophaga BN6

Andreas Keck1,{dagger}, Doris Conradt1, Anette Mahler1, Andreas Stolz2, Ralf Mattes1 and Joachim Klein1,{ddagger}

1 Institut für Industrielle Genetik, Universität Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany
2 Institut für Mikrobiologie, Universität Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany

Correspondence
Joachim Klein
joachim.klein{at}lonza.com

Sphingomonas xenophaga BN6 degrades various (substituted) naphthalenesulfonates to the corresponding (substituted) salicylates. A gene cluster was identified on the plasmid pBN6 which coded for several enzymes participating in the degradative pathway for naphthalenesulfonates. A DNA fragment of 16 915 bp was sequenced which contained 17 ORFs. The genes encoding the 1,2-dihydroxynaphthalene dioxygenase, 2-hydroxychromene-2-carboxylate isomerase, and 2'-hydroxybenzalpyruvate aldolase of the naphthalenesulfonate pathway were identified on the DNA fragment and the encoded proteins heterologously expressed in Escherichia coli. Also, the genes encoding the ferredoxin and ferredoxin reductase of a multi-component, ring-hydroxylating naphthalenesulfonate dioxygenase were identified by insertional inactivation. The identified genes generally demonstrated the highest degree of homology to enzymes encoded by the phenanthrene-degrading organism Sphingomonas sp. P2, or the megaplasmid pNL1 of the naphthalene- and biphenyl-degrading strain Sphingomonas aromaticivorans F199. The genes of S. xenophaga BN6 participating in the degradation of naphthalenesulfonates also shared the same organization in three different transcriptional units as the genes involved in the degradation of naphthalene, biphenyl, and phenanthrene previously found in Sphingomonas sp. P2 and S. aromaticivorans F199. The genes were flanked in S. xenophaga BN6 by ORFs which specify proteins that show the highest homologies to proteins of mobile genetic elements.

Abbreviations: DHNDO, 1,2-dihydroxynaphthalene dioxygenase; 2NSA, naphthalene-2-sulfonic acid; NSA, naphthalenesulfonic acid; NsaA, naphthalenesulfonate dioxygenase; NsaC, 1,2-dihydroxynaphthalene dioxygenase; NsaD, 2-hydroxychromene-2-carboxylate isomerase; NsaE, 2'-hydroxybenzalpyruvate aldolase; NsaF, salicylaldehyde dehydrogenase

The GenBank/EMBL/DDBJ accession number for the sequence determined in this work is U65001.

{dagger}Present address: Deutsches Ressourcenzentrum für Genomforschung, Heubnerweg 6, 14059 Berlin, Germany.

{ddagger}Present address: Lonza AG, Biotechnology Research and Development, CH-3930 Visp, Switzerland.






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