Gene cloning and in vivo characterization of a dibenzothiophene dioxygenase from Xanthobacter polyaromaticivorans.
Xanthobacter polyaromaticivorans sp. nov. 127W is a bacterial strain that is capable of degrading a wide range of cyclic aromatic compounds such as dibenzothiophene, biphenyl, naphthalene, anthracene, and phenanthrene even under extremely low oxygen [dissolved oxygen (DO)< or = 0.2 ppm] conditions (Hirano et al., Biosci Biotechnol Biochem 68:557-564, 2004). A major protein fraction carrying dibenzothiophene degradation activity was purified. Based on its partial amino acid sequences, dbdCa gene encoding alpha subunit terminal oxygenase (DbdCa) and its flanking region were cloned and sequenced. A phylogenetic analysis based on the amino acid sequence demonstrates that DbdCa is a member of a terminal oxygenase component of group IV ring-hydroxylating dioxygenases for biphenyls and monocyclic aromatic hydrocarbons, rather than group III dioxygenases for polycyclic aromatic hydrocarbons. Gene disruption in dbdCa abolished almost of the degradation activity against biphenyl, dibenzothiophene, and anthracene. The gene disruption also impaired degradation activity of the strain under extremely low oxygen conditions (DO< or = 0.2 ppm). These results indicate that Dbd from 127W represents a group IV dioxygenase that is functional even under extremely low oxygen conditions.
Hirano
Shin-Ichi
S
Department of Material and Life Science, Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan.
Haruki
Mitsuru
M
Takano
Kazufumi
K
Imanaka
Tadayuki
T
Morikawa
Masaaki
M
Kanaya
Shigenori
S
D000595:Amino Acid Sequence; D000873:Anthracenes / Q000378:metabolism; D003001:Cloning, Molecular; D019976:Cloning, Organism; D049308:Dioxygenases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D017353:Gene Deletion; D023061:Gene Order; D006841:Hydrocarbons, Aromatic / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010802:Phylogeny; D021122:Protein Subunits / Q000235:genetics; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013876:Thiophenes / Q000378:metabolism*; D020584:Xanthobacter / Q000201:enzymology* / Q000235:genetics
D000873:Anthracenes; D006841:Hydrocarbons, Aromatic; D021122:Protein Subunits; D013876:Thiophenes; D049308:Dioxygenases; C034020:anthracene; C016366:dibenzothiophene