The bphC gene-encoded 2,3-dihydroxybiphenyl-1,2-dioxygenase is involved in complete degradation of dibenzofuran by the biphenyl-degrading bacterium Ralstonia sp. SBUG 290.
OBJECTIVE
Biphenyl-degrading bacteria are able to metabolize dibenzofuran via lateral dioxygenation and meta-cleavage of the dihydroxylated dibenzofuran produced. This degradation was considered to be incomplete because accumulation of a yellow-orange ring-cleavage product was observed. In this study, we want to characterize the 1,2-dihydroxydibenzofuran cleaving enzyme which is involved in dibenzofuran degradation in the bacterium Ralstonia sp. SBUG 290.


RESULTS
In this strain, complete degradation of dibenzofuran was observed after cultivation on biphenyl. The enzyme shows a wide substrate utilization spectrum, including 1,2-dihydroxydibenzofuran, 2,3-dihydroxybiphenyl, 1,2-dihydroxynaphthalene, 3- and 4-methylcatechol and catechol. MALDI-TOF analysis of the protein revealed a strong homology to the bphC gene products. We therefore cloned a 3.2 kb DNA fragment containing the bphC gene of Ralstonia sp. SBUG 290. The deduced amino acid sequence of bphC is identical to that of the corresponding gene in Pseudomonas sp. KKS102. The bphC gene was expressed in Escherichia coli and the meta-fission activity was detected using either 2,3-dihydroxybiphenyl or 1,2-dihydroxydibenzofuran as substrate.


CONCLUSIONS
These results demonstrate that complete degradation of dibenzofuran by biphenyl degraders can occur after initial oxidation steps catalysed by gene products encoded by the bph-operon. The ring fission of 1,2-dihydroxydibenzofuran is catalysed by BphC. Differences found in the metabolism of the ring fission product of dibenzofuran among biphenyl degrading bacteria are assumed to be caused by different substrate specificities of BphD.


CONCLUSIONS
This study shows for the first time that the gene products of the bph-operon are involved in the mineralization of dibenzofuran in biphenyl degrading bacteria.
Wesche
J
J
Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, Greifswald, Germany. wesche@uni-greifswald.de
Hammer
E
E
Becher
D
D
Burchhardt
G
G
Schauer
F
F
D000595:Amino Acid Sequence; D001572:Benzofurans / Q000378:metabolism*; D001673:Biodegradation, Environmental; D017124:Conserved Sequence; D049308:Dioxygenases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli; D015870:Gene Expression; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D043367:Ralstonia / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D012988:Soil Microbiology
D001572:Benzofurans; C023614:dibenzofuran; D049308:Dioxygenases; C074272:2,3-dihydroxybiphenyl oxygenase