Characterization of the desulfurization genes from Rhodococcus sp. strain IGTS8.
Rhodococcus sp. strain IGTS8 possesses an enzymatic pathway that can remove covalently bound sulfur from dibenzothiophene (DBT) without breaking carbon-carbon bonds. The DNA sequence of a 4.0-kb BstBI-BsiWI fragment that carries the genes for this pathway was determined. Frameshift and deletion mutations established that three open reading frames were required for DBT desulfurization, and the genes were designated soxABC (for sulfur oxidation). Each sox gene was subcloned independently and expressed in Escherichia coli MZ1 under control of the inducible lambda pL promoter with a lambda cII ribosomal binding site. SoxC is an approximately 45-kDa protein that oxidizes DBT to DBT-5,5'-dioxide. SoxA is an approximately 50-kDa protein responsible for metabolizing DBT-5,5'-dioxide to an unidentified intermediate. SoxB is an approximately 40-kDa protein that, together with the SoxA protein, completes the desulfurization of DBT-5,5'-dioxide to 2-hydroxybiphenyl. Protein sequence comparisons revealed that the predicted SoxC protein is similar to members of the acyl coenzyme A dehydrogenase family but that the SoxA and SoxB proteins have no significant identities to other known proteins. The sox genes are plasmidborne and appear to be expressed as an operon in Rhodococcus sp. strain IGTS8 and in E. coli.
Department of Microbiology and Immunology, University of North Dakota School of Medicine, Grand Forks 58202.
S A
SA
Denome
C
C
Oldfield
L J
LJ
Nash
K D
KD
Young
D000595:Amino Acid Sequence; D001483:Base Sequence; D001673:Biodegradation, Environmental; D001713:Biphenyl Compounds / Q000378:metabolism; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D044943:Fatty Acid Desaturases / Q000235:genetics; D016368:Frameshift Mutation; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D012240:Rhodococcus / Q000201:enzymology / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013455:Sulfur / Q000378:metabolism*; D013876:Thiophenes / Q000378:metabolism*
D001713:Biphenyl Compounds; D003574:Cytochrome c Group; D011994:Recombinant Proteins; D013876:Thiophenes; C090101:dibenzothiophene-5,5-dioxide; D013455:Sulfur; C004369:2-phenylphenol; D010088:Oxidoreductases; D044943:Fatty Acid Desaturases; C090100:dibenzothiophene monooxygenase; C090099:diheme cytochrome c; C064500:flavocytochrome c sulfide dehydrogenase; C016366:dibenzothiophene