Elucidation of the metabolic pathway for dibenzothiophene desulphurization by Rhodococcus sp. strain IGTS8 (ATCC 53968).
Rhodococcus sp. strain IGTS8 (ATCC 53968) is able to utilize dibenzothiophene (DBT) as a sole source of sulphur. The carbon skeleton of DBT is not metabolized and is conserved as 2-hydroxybiphenyl (HBP), which accumulates in the medium. This phenotype is due to the expression of the plasmid-encoded DBT-desulphurization (dsz) operon, which encodes three proteins, DszA, B and C. In this paper it is shown, using [35S]DBT radiolabelling studies, that sulphur is released in the form of inorganic sulphite. The pathway of DBT desulphurization is described in detail. In summary, DszC catalyses the stepwise S-oxidation of DBT, first to dibenzothiophene 5-oxide (DBTO) and then to dibenzothiophene 5,5-dioxide (DBTO2); DszA catalyses the conversion of DBTO2 to 2-(2'-hydroxyphenyl)benzene sulphinate (HBPSi-) and DszB catalyses the desulphination of HBPSi- to give HBP and sulphite. Studies with cell-free extracts show that DszA and DszC, but not DszB, require NADH for activity. 18O2-labelling studies show that each incorporated oxygen atom is derived directly from molecular oxygen. These results are consistent with the role of DszC as a mono-oxygenase, of DszA as an apparently unique enzyme which catalyses the reductive hydroxylation of DBTO2 leading to cleavage of the thiophene ring, and of DszB as an aromatic sulphinic acid hydrolase.
Oldfield
Christopher
C
Pogrebinsky
Olga
O
Energy Biosystems Corporation, 4200 Research Forest Drive, The Woodlands, TX 77381, USA.
Simmonds
Julie
J
Energy Biosystems Corporation, 4200 Research Forest Drive, The Woodlands, TX 77381, USA.
Olson
Edwin S
ES
Energy and Environmental Research Center, University of North Dakota, Grand Forks, ND 58202, USA.
Kulpa
Charles F
CF
Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556-0309, USA.
D015258:Acid Rain / Q000517:prevention & control; D001673:Biodegradation, Environmental; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D009876:Operon; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D011995:Recombination, Genetic; D012240:Rhodococcus / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D013447:Sulfites / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism; D013876:Thiophenes / Q000378:metabolism*
D015258:Acid Rain; D013447:Sulfites; D013876:Thiophenes; D009243:NAD; D013455:Sulfur; D010088:Oxidoreductases; C090100:dibenzothiophene monooxygenase; C016366:dibenzothiophene