Desulphurization of dibenzothiophene by bacteria

Four out of 187 strains, from enrichment cultures of dibenzothiophene (DBT), grew on DBT or thiophene 2-carboxylate as S sources. The four isolates, presumptively identified as Agrobacterium sp., Xanthomonas sp. and Corynebacterium spp., individually and together desulphurized DBT, producing 2-hydroxybiphenyl and sulphate.M. Constanti and A. Bordons are with the Departament de Bioquímica i Biotecnologia, and J. Giralt is with the Departament d'Enginyeria Química, both of the Universitat Rovira i Virgili, Pl. Imperial Tarraco 1, 43005 Tarragona, Catatonia, Spain     

Degradation of phenanthrene,methylphenanthrenes and dibenzothiophene by a Sphingomonas strain 2mpII

Strain Sphingomonassp. (2MPII), isolated from marine sediment, was able to utilize phenanthrene (P) or 2-methylphenanthrene (2MP) as the sole carbon source. However, 9-methylphenanthrene (9MP) and dibenzothiophene (DBT) were weakly degraded. The degradation rates of 9MP and DBT increased in the presence of 2MP, whilst the degradation rate of 2MP increased in the presence of 9MP. However, the presence of DBT inhibited the degradation of 2MP. DBT sulfone, a DBT metabolite, was not assimilated by the bacteria and its presence also decreased the degradation rate of 2MP.     

Combining co-culturing of Paenibacillus strains and Vitreoscilla hemoglobin expression as a strategy to improve biodesulfurization

Enhancement of the desulfurization activities of Paenibacillus strains 32O-W and 32O-Y were investigated using dibenzothiophene (DBT) and DBT sulfone (DBTS) as sources of sulphur in growth experiments. Strains 32O-W, 32O-Y and their co-culture (32O-W plus 32O-Y), and Vitreoscilla hemoglobin (VHb) expressing recombinant strain 32O-Yvgb and its co-culture with strain 32O-W were grown at varying concentrations (0·1–2 mmol l⁻¹) of DBT or DBTS for 96 h, and desulfurization measured by production of 2-hydroxybiphenyl (2-HBP) and disappearance of DBT or DBTS. Of the four cultures grown with DBT as sulphur source, the best growth occurred for the 32O-Yvgb plus 32O-W co-culture at 0·1 and 0·5 mmol l⁻¹ DBT. Although the presence of vgb provided no consistent advantage regarding growth on DBTS, strain 32O-W, as predicted by previous work, was shown to contain a partial 4S desulfurization pathway allowing it to metabolize this 4S pathway intermediate.     

Degradation of dibenzothiophene by Pseudomonas putida

The microbial degradation of dibenzothiophene (DBT) and other organosulphur compounds such as thiophene-2-carboxylate (T2C) is of interest for the potential desulphurization of coal. The feasibility of degradation of DBT and T2C by Pseudomonas putida and other bacteria was analysed. Pseudomonas putida oxidized sulphur from DBT in the presence of yeast extract, but it did not when DBT was the sole source of carbon.     

Plasmid-mediated degradation of dibenzothiophene by Pseudomonas species

The microbial transformation of dibenzothiophene (DBT) is of interest in the potential desulfurization of oil. We isolated three soil Pseudomonas species which oxidized DBT to characteristic water-soluble, sulfur-containing products. Two of our isolates harbored a 55-megadalton plasmid; growth in the presence of novobiocin resulted in both loss of the plasmid and loss of the ability to oxidize DBT. Reintroduction of the plasmid restored the ability to oxidize DBT to water-soluble products. The products resulting from the oxidation of DBT were characterized and included 3-hydroxy-2-formyl benzothiophene, 3-oxo-[3'-hydroxy-thionaphthenyl-(2)-methylene]-dihydrothionaph thene, and the hemiacetal and trans forms of 4-[2-(3-hydroxy)-thianaphthenyl]-2-oxo-3-butenoic acid. The products of DBT oxidation were inhibitory to cell growth and further DBT oxidation. DBT oxidation in our soil isolates was induced by naphthalene or salicylate and to a much lesser extent by DBT and was repressed by succinate.     

Plasmid-mediated degradation of dibenzothiophene by Pseudomonas species.

The microbial transformation of dibenzothiophene (DBT) is of interest in the potential desulfurization of oil. We isolated three soil Pseudomonas species which oxidized DBT to characteristic water-soluble, sulfur-containing products. Two of our isolates harbored a 55-megadalton plasmid; growth in the presence of novobiocin resulted in both loss of the plasmid and loss of the ability to oxidize DBT. Reintroduction of the plasmid restored the ability to oxidize DBT to water-soluble products. The products resulting from the oxidation of DBT were characterized and included 3-hydroxy-2-formyl benzothiophene, 3-oxo-[3'-hydroxy-thionaphthenyl-(2)-methylene]-dihydrothionaph thene, and the hemiacetal and trans forms of 4-[2-(3-hydroxy)-thianaphthenyl]-2-oxo-3-butenoic acid. The products of DBT oxidation were inhibitory to cell growth and further DBT oxidation. DBT oxidation in our soil isolates was induced by naphthalene or salicylate and to a much lesser extent by DBT and was repressed by succinate.     

Conservation of plasmid-encoded dibenzothiophene desulfurization genes in several rhodococci.

The cloned sulfur oxidation (desulfurization) genes (sox) for dibenzothiophene (DBT) from the prototype Rhodococcus sp. strain IGTS8 were used in Southern hybridization and PCR experiments to establish the DNA relatedness in six new rhodococcal isolates which are capable of utilizing DBT as a sole sulfur source for growth. The ability of these strains to desulfurize appears to be an exclusive property of a 4-kb gene locus on a large plasmid of ca. 150 kb in IGTS8 and ca. 100 kb in the other strains. Besides a difference in plasmid profile, IGTS8 is distinguishable from the other strains in at least the copy number of the insertion sequence IS1166, which is associated with the sox genes.     

Microbial desulfurization of dibenzothiophene in the presence of hydrocarbon

The bacterium, Rhodococcus erythropolis H-2, which can utilize dibenzothiophene (DBT) as a sole source of sulfur in the presence of hydrocarbon, was isolated from soil samples. When this strain was cultivated in a medium containing 0.27 mM DBT and 40% n-tetradecane, DBT was metabolized stoichiometrically to 2-hydroxybiphenyl within 1 day. This strain grew in the presence of n-octane and longer-carbonchain hydrocarbons, but not with n-hexane, styrene, p-xylene, cyclooctane or toluene. DBT degradation proceeded in the resting cell system with lyophilized cells of this strain. The addition of n-tetradecane enhanced the reaction rate, the optimal concentration being 40%. DBT degradation occurred in the reaction mixture even in the presence of 70% n-tetradecane, whereas at concentrations above 80% n-tetradecane suppressed the degradation.     

Degradation of dibenzothiophene by Brevibacterium sp.DO

Dibenzothiophene, a polycyclic aromatic sulfur heterocycle, represents as a model compound the organic sulfur integrated in the macromolecular coal matrix. A pure culture of a Brevibacterium species was isolated, which is able to use dibenzothiophene as sole source of carbon, sulfur and energy for growth. During dibenzothiophene utilization sulfite was released in a stoichiometrical amount and was further oxidized to sulfate. Three metabolites of dibenzothiophene degradation were isolated and identified as dibenzothiophene-5-oxide, dibenzothiophene-5-dioxide and benzoate by cochromatography, UV spectroscopy and gas chromatographymass spectrometry analyses. Based on the identified metabolites a pathway for the degradation of dibenzothiophene by Brevibacterium sp. DO is proposed.Non-standard abbreviations DBT dibenzothiophene - PASH polycyclic aromatic sulfur heterocycle - PAH polycyclic aromatic hydrocarbons - GC-MS gas chromatography-mass spectrometry - HPLC high pressure liquid chromatography - IC ion chromatography     

Desulfurization of dibenzothiophene by Corynebacterium sp. strain SY1.

Strain SY1, identified as a Corynebacterium sp., was isolated on the basis of the ability to utilize dibenzothiophene (DBT) as a sole source of sulfur. Strain SY1 could utilize a wide range of organic and inorganic sulfur compounds, such as DBT sulfone, dimethyl sulfide, dimethyl sulfoxide, dimethyl sulfone, CS2, FeS2, and even elemental sulfur. Strain SY1 metabolized DBT to dibenzothiophene-5-oxide, DBT sulfone, and 2-hydroxybiphenyl, which was subsequently nitrated to produce at least two different hydroxynitrobiphenyls during cultivation. These metabolites were separated by silica gel column chromatography and identified by nuclear magnetic resonance, UV, and mass spectral techniques. Resting cells of SY1 desulfurized toluenesulfonic acid and released sulfite anion. On the basis of these results, a new DBT degradation pathway is proposed.     

Degradation of dibenzothiophene and its metabolite 3-hydroxy-2-formylbenzothiophene by an environmental isolate

Microbial degradation of dibenzothiophene (DBT) beyond 3-hydroxy-2-formylbenzothiophene (HFBT), a commonly detected metabolite of the Kodama pathway for DBT metabolism, and the catabolic intermediates leading to its mineralization are not fully understood. The enrichment cultures cultivated from crude oil contaminated soil led to isolation of ERI-11; a natural mixed culture, selected for its ability to deplete DBT in basal salt medium (BSM). A bacterial strain isolated from ERI-11, and tentatively named A₁₁, degraded more than 90 % of the initial DBT (270 µM), present as the sole carbon and sulfur source, in 72 h. Gas chromatography–mass spectrophotometry (GC–MS) analyses of the DBT degrading A₁₁ culture medium extracts led to detection of HFBT. The metabolite HFBT, produced using A₁₁, was used in degradation assays to evaluate its metabolism by the bacteria isolated in this study. Ultra violet–visible spectrophotometry and high-performance liquid chromatography analyses established the ability of the strain A₁₁ to deplete HFBT, present as the sole sulfur and carbon source in BSM. GC–MS analyses showed the presence of 2-mercaptobenzoic acid in the HFBT degrading A₁₁ culture extracts. The findings in this study establish that the environmental isolate A₁₁ possesses the metabolic capacity to degrade DBT beyond the metabolite HFBT. The compound 2-mercaptobenzoic acid is an intermediate formed on HFBT degradation by A₁₁.     

Desulfurization of dibenzothiophene by Corynebacterium sp. strain SY1.

Strain SY1, identified as a Corynebacterium sp., was isolated on the basis of the ability to utilize dibenzothiophene (DBT) as a sole source of sulfur. Strain SY1 could utilize a wide range of organic and inorganic sulfur compounds, such as DBT sulfone, dimethyl sulfide, dimethyl sulfoxide, dimethyl sulfone, CS2, FeS2, and even elemental sulfur. Strain SY1 metabolized DBT to dibenzothiophene-5-oxide, DBT sulfone, and 2-hydroxybiphenyl, which was subsequently nitrated to produce at least two different hydroxynitrobiphenyls during cultivation. These metabolites were separated by silica gel column chromatography and identified by nuclear magnetic resonance, UV, and mass spectral techniques. Resting cells of SY1 desulfurized toluenesulfonic acid and released sulfite anion. On the basis of these results, a new DBT degradation pathway is proposed.     

Microbial oxidation of dibenzothiophene by the thermophilic organism Sulfolobus acidocaldarius

The refractory organic sulfur compound dibenzothiophene (DBT) has been oxidized by the thermophilic, sulfur oxidizing organism Sulfolobus acidocaldarius. Sulfate ions were released into the medium as the oxidation product. The kinetics of this oxidation have been investigated on the basis of sulfate released as a result of oxidation. Dibenzothiophene was found to be inhibitory to the organisms for initial concentrations over 500 mg/L. The organism may prove to be capable of oxidizing thiophene compounds present in oil refinery wastewater, coal, and crude oil.     

The effect ofn-alkanes in the degradation of dibenzothiophene and of organic sulfur compounds in heavy oil by aPseudomonas sp.

Summary The microbial degradation of organic sulfur compounds was examined in aerobic conditions employing a pure culture of aPseudomonas sp., isolated from the soil. The effect ofn-alkanes on the degradation of dibenzothiophene (DBT) showed that the assimilation of the sulfur compound by the microorganism is favoured byn-dodecane. Moreover, the saturated fraction was seen to enhance the degradation of the sulfur compounds to be found in a deasphaltenated heavy oil.     

Influence of incubation period on the plasmid content of proliferating Nocardioides sp.

S. SANDHYA. 1996. Nocardioides sp., isolated for organic sulphur degradation, harbours catabolic plasmid pSB1 of molecular weight 34.2 kb. A correlation was noticed between the plasmid content and degradation of the organic sulphur compound dibenzothiophene (DBT). The maximum oxidation of DBT was only after 64 h, during which the content of plasmid DNA was also optimum.     

Conversion of dibenzothiophene to biphenyl by sulfate-reducing bacteria isolated from oil field production facilities

Summary Numerous purified and characterized sulfate-reducing bacteria and bacterial communities isolated from oil-field production facilities were shown to convert dibenzothiophene (DBT) into biphenyl (BP). The maximum degree of conversion to biphenyl was 1.14 % and 0.39 % for purified sulfate-reducing bacteria and community bacteria, respectively. The purified sulfate-reducing bacteria and bacterial communities were identified by 16S rRNA ribotyping.     

Oxidation of dibenzothiophene (DBT) by Serratia marcescens UCP 1549 formed biphenyl as final product

ABSTRACT: BACKGROUND: The desulphurization of dibenzothiophene (DBT), a recalcitrant thiophenic fossil fuel component by Serratia marcescens (UCP 1549) in order for reducing the sulphur content was investigated. The study was carried out establishing the growth profile using Luria Bertani medium to different concentrations of DBT during 120hours at 28oC, and orbital shaker at 150rpm. RESULTS: The results indicated that concentrations of DBT 0.5, 1.0 and 2.0 mM do not affected the growth of the bacterium. The DBT showed similar Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MCB) (3.68 mM). The desulphurization of DBT by S. marcescens was used with 96 hours of growth on 2mM of DBT, and was determined by gas chromatography (GC) and GC-mass spectrometry. In order to study the desulphurization process by S. marcescens was observed the presence of a sulfur-free product at 16 hours of cultivation The results show that S. marcescens oxidizes DBT to its corresponding DBT-5 oxide and then to DBT-sulfone, without the formation of any biphenyl. CONCLUSIONS: The data suggests the use of metabolic pathway "4S" by S. marcescens (UCP 1549) and formed biphenyl. The microbial desulphurization process by S. Serratia can be suggest significant reducing sulphur content in DBT, and showed promising potential for reduction of the sulfur content in diesel oil.     

Biodegradation of dibenzothiophene by thermophilic bacteria

Anaerobic microbial biodegradation of dibenzothiophene (DBT) was studied using thermophilic bacteria obtained from crude oil. A mixed culture was obtained that degraded 98% of DBT at 0.5 mg ml^–1 at 65 °C over 15 days both in the presence and in the absence of Methyl Viologen.     

Oxidation of dibenzothiophene byCunninghamella elegans

The metabolism of dibenzothiophene (DBT) by the catabolically versatile fungusCunninghamella elegans (ATCC 36112) was investigated. This fungus has been previously shown to metabolize polycyclic aromatic hydrocarbons including nitropolycyclic aromatics, but has not been shown to metabolize DBT. The results show thatC. elegans oxidizes DBT to its corresponding DBT-5 oxide and then to DBT-sulfone, without the formation of any biphenyl intermediates.Paper from the Idaho Agricultural Experiment Station.     

Purification, characterization and crystallization of enzymes for dibenzothiophene desulfurization

DszC and DszA, DBT monooxygenase and DBT sulfone monooxygenase, respectively, involved in dibenzothiophene (DBT) desulfurization, were purified to homogeneity from Rhodococcus erythropolis D-1. The two enzymes were crystallized and enzymologically characterized. We found a high activity of flavin reductase in the non-DBT-desulfurizing bacterium, Paenibacillus polymyxa A-1, which is essential for DszC and A activities, and purified to homogeneity and characterized the enzyme.