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1.
A yeast strain AEH was isolated from oil contaminated soil and identified by analysis of 18S and 26S ribosomal DNA sequences asPichia anomala. Strain AEH was capable of degrading naphthalene, phenanthrene and chrysene, singly, and benzo(a)pyrene in combination. The yeast degraded 5.36 mg naphthalene l?1 within 2 days, and 5.04 mg phenanthrene l?1 and 1.54 mg chrysene 1?1 within 10 days. When a mixture of the four polycyclic aromatic hydrocarbons (PAHs) was treated at a concentration between 2.98 mg l?1 and 6.89 mg l?1, degradation rates were delayed for naphthalene and phenanthrene (3.79 mg l?1 and, 4.20 mg l?1 within 10 days, respectively), but enhanced for chrysene and benzo(a)pyrene (3.37 mg l?1 and, 1.91 mg l?1 within 10 days, respectively). In a binary system, all of the other 3 PAHs could be utilized as the carbon source for the cometabolic degradation of benzo(a)pyrene with naphthale ne as the best one.  相似文献   

2.
In order to study the mechanisms regulating the phenanthrene degradation pathway and the intermediate-metabolite accumulation in strain S. paucimobilis 20006FA, we sequenced the genome and compared the genome-based predictions to experimental proteomic analyses. Physiological studies indicated that the degradation involved the salicylate and protocatechuate pathways, reaching 56.3% after 15 days. Furthermore, the strain degraded other polycyclic aromatic hydrocarbons (PAH) such as anthracene (13.1%), dibenzothiophene (76.3%), and fluoranthene. The intermediate metabolite 1-hydroxy-2-naphthoic acid (HNA) accumulated during phenanthrene catabolism and inhibited both bacterial growth and phenanthrene degradation, but exogenous-HNA addition did not affect further degradation. Genomic analysis predicted 126 putative genes encoding enzymes for all the steps of phenanthrene degradation, which loci could also participate in the metabolism of other PAH. Proteomic analysis identified enzymes involved in 19 of the 23 steps needed for the transformation of phenanthrene to trichloroacetic-acid intermediates that were upregulated in phenanthrene cultures relative to the levels in glucose cultures. Moreover, the protein-induction pattern was temporal, varying between 24 and 96 h during phenanthrene degradation, with most catabolic proteins being overexpressed at 96 h—e. g., the biphenyl dioxygenase and a multispecies (2Fe–2S)-binding protein. These results provided the first clues about regulation of expression of phenanthrene degradative enzymes in strain 20006FA and enabled an elucidation of the metabolic pathway utilized by the bacterium. To our knowledge the present work represents the first investigation of genomic, proteomic, and physiological studies of a PAH-degrading Sphingomonas strain.  相似文献   

3.
A bacterial strain able to grow in pure culture with chrysene as sole added carbon and energy source was isolated from PAH-contaminated soil after successive enrichment cultures in a biphasic growth medium. Initially, growth occurred in the form of a biofilm at the interface between the aqueous and non-aqueous liquid phases. However, after a certain time, a transition occurred in the enrichment cultures, with growth occurring in suspension and a concomitant increase in the rate of chrysene degradation. The strain responsible for chrysene degradation in these cultures, named Sphingomonas sp. CHY-1, was identified by 16S rDNA sequencing as a novel sphingomonad, the closest relative in the databases being Sphingomonas xenophaga BN6T (96% sequence identity). Both these strains clustered with members of the genera Sphingobium and Rhizomonas, but could not be categorically assigned to either genus. Sphingomonas sp. CHY-1 was characterized in terms of its growth on chrysene and other PAH, and the kinetics of chrysene degradation and 14C-chrysene mineralization were measured. At an initial chrysene concentration of 0.5 g l(-1) silicone oil, and an organic/aqueous phase ratio of 1:4, chrysene was 50% degraded after 5 days incubation and 97.5% degraded after 35 days. The protein content of cultures reached a maximum value of 11.5 microg ml(-1) aqueous phase, corresponding to 92 mg g(-1) chrysene. 14C-labelled chrysene was 50% mineralized after 6-8 weeks incubation, 10.7% of the radioactivity was incorporated into cell biomass and 8.4% was found in the aqueous culture supernatant. Sphingomonas sp. CHY-1 also grew on naphthalene, phenanthrene and anthracene, and naphthalene was the preferred substrate, with a doubling time of 6.9 h.  相似文献   

4.

Objectives

With the view of designing a single biocatalyst for biorefining, carbazole dioxygenase was cloned from Pseudomonas sp. and expressed in Rhodococcus sp.

Results

The recombinant, IGTS8, degraded both carbazole and dibenzothiophene at 400 mg/l in 24 h. Maximum carbazole degradation was in 1:1 (v/v) hexadecane/aqueous phase. Anthracene, phenanthrene, pyrene, fluoranthene and fluorine were also degraded without affecting the aliphatic component.

Conclusions

Recombinant Rhodococcus sp. IGTS8 can function as a single biocatalyst for removing major contaminants of fossil fuels viz. dibenzothiophene, carbazole and polyaromatic compounds.
  相似文献   

5.
A mixture of six polyaromatic hydrocarbons (naphthalene, phenanthrene, fluoranthene, pyrene, chyrysene and benzo[a]pyrene), varying in size from 2 to 5 rings, was dissolved in dodecane, and used as the delivery phase of a partitioning bioreactor. Two species of Sphingomonas were then used individually, and as a consortium, to determine which of the PAHs were degraded. Only low molecular weight PAHs (naphthalene, phenanthrene and fluoranthene) were degraded by the individual strains, but the consortium degraded all substrates either to completion or near completion.  相似文献   

6.
Mycobacterium PYR-1 was used in a two-phase partitioning bioreactor (TPPB) to degrade low and high molecular weight polycyclic aromatic hydrocarbons. TPPBs are characterized by a cell-containing aqueous phase, and an immiscible and biocompatible organic phase that partitions toxic substrates to the cells based on their metabolic demand and the thermodynamic equilibrium of the system. A bioavailable solvent, that is, a solvent usable as a carbon source, was used as the organic layer. Although bioavailable solvents are traditionally deemed unsuitable for use in TPPBs, bis(ethylhexyl) sebacate had superior chemical properties to other solvents examined and was cost-effective. In this system, 1 g of phenanthrene and 1 g of pyrene were completely degraded within 4 days, at rates of 168 mg l(-1) day(-1) and 138 mg l(-1 )day(-1), respectively, based on a 3-l aqueous volume. This is the highest pyrene degradation rate reported in the literature to date. Significant degradation of naphthalene and anthracene was also obtained. This work demonstrates that bioavailable solvents can be successfully used in TPPB systems, and may change the protocols commonly used to select solvents for TPPBs in the future.  相似文献   

7.
A two-phase partitioning bioreactor (TPPB) utilizing the bacterium Sphingomonas aromaticivorans B0695 was used to degrade four low molecular weight (LMW) polycyclic aromatic hydrocarbons (PAHs). The TPPB concept is based on the use of a biocompatible, immiscible organic solvent in which high concentrations of recalcitrant substrates are dissolved. These substances partition into the cell-containing aqueous phase at rates determined by the metabolic activity of the cells. Experiments showed that the selected solvent, dodecane, could be successfully used in both solvent extraction experiments (to remove PAHs from soil) and in a TPPB application. Further testing demonstrated that solvent extraction from spiked soil was enhanced when a solvent combination (dodecane and ethanol) was used, and it was shown that the co-solvent did not significantly affect TPPB performance. The TPPB achieved complete biodegradation of naphthalene, phenanthrene, acenaphthene and anthracene at a volumetric consumption rate of 90 mg l(-1) h(-1) in approximately 30 h. Additionally, a total of 20.0 g of LMW PAHs (naphthalene and phenanthrene) were biodegraded at an overall volumetric rate of 98 mg l(-1) h(-1) in less than 75 h. Degradation rates achieved using the TPPB and S. aromaticivorans B0695 are much greater than any others previously reported for an ex situ PAH biodegradation system operating with a single species.  相似文献   

8.
White shrimp Litopenaeus vannamei (Boone) held in 35 per thousand seawater were challenged with Vibrio alginolyticus at a dose of 3 x 10(5) colony-forming units (cfu) shrimp(-1), and then placed in water containing concentrations of Cu2+ at 0 (control), 1, 5, 10 and 20 mg l(-1). Mortality of shrimp in 5, 10 and 20 mg l(-1) Cu2+ was significantly higher than those in 1 mg l(-1) Cu2+ and the control solution after 24-96 h. In another experiment, L. vannamei which had been exposed to control, 1, 5, 10 and 20 mg l(-1) Cu2+ for 24, 48 and 96 h were examined for THC (total haemocyte count), phenoloxidase activity, respiratory burst (release of superoxide anion), phagocytic activity and clearance efficiency to V. alginolyticus. Copper concentrations at 1 mg l(-1) or greater for 24h resulted in decreased THC, phenoloxidase activity, phagocytic activity and clearance efficiency, whereas copper concentration at 20 mg l(-1) caused significant increase in respiratory burst of L. vannamei. In conclusion, concentration of Cu2+ at 1 mg l(-1) or greater increased the susceptibility of L. vannamei to V. alginolyticus infection by a depression in immune ability. The release of superoxide anion by L. vannamei exposed to 20 mg l(-1) Cu2+ was considered to be cytotoxic to the host.  相似文献   

9.
Beijerinckia mobilis 1f capable of degrading polycyclic aromatic hydrocarbons (PAHs) was isolated from a soil contaminated with creosote. Strain 1f could utilize phenanthrene and naphthalene as the sole sources of carbon. The mean rate of phenanthrene degradation during culture growth was 7-8 micrograms/(ml h). After cultivation under nonselective conditions, strain 1f retained its ability to degrade phenanthrene. Cometabolism considerably widened the range of PAHs that could be transformed by strain 1f. The strain was able to grow in a mineral medium with creosote as the sole source of carbon. After 30 days of cultivation in this medium, the total concentration of PAHs decreased from 665.5 mg/l to 170 mg/l.  相似文献   

10.
A newly isolated Rhodococcus sp. strain p52 could aerobically utilize dibenzofuran as the sole source of carbon and energy, and completely remove dibenzofuran at 500 mg?l?1 within 48 h. The strain metabolizes dibenzofuran by initial angular dioxygenation to yield 2,2′,3-trihydroxybiphenyl. Strain p52 could also remove 70 % of 100 mg?l?1 2-chlorodibenzofuran within 96 h and could metabolize a variety of aromatic compounds, namely dibenzo-p-dioxin, 2,8-dichlorodibenzofuran, dibenzothiophene, biphenyl, naphthalene, fluorene, phenanthrene, anthracene, carbazole, indole, xanthene, phenoxathiin, xanthone, and 9-fluorenone. Two distinct gene clusters encoding angular dioxygenases (DbfA and DfdA) were amplified and sequenced. The dbfA and dfdA gene clusters are located on two circular plasmids, pDF01 and pDF02, respectively. Both plasmids are self-transmissible; that is, they can transfer to the Gram-positive bacterium Bacillus cereus by conjugation.  相似文献   

11.
The mixed bacterial culture MK1 was capable of degrading a wide spectrum of aromatic compounds both as free and as immobilized cells. By offering anthracene oil or a defined mixture of phenol, naphthalene, phenanthrene, anthracene and pyrene (in concentrations of 0.1–0.2 mm, respectively) as sources of carbon and energy, a specific degradation pattern correlating with the condensation degree was observed. Regarding the defined mixture of aromatic hydrocarbons, complete metabolism was reached for phenol (0.1 mm) after 1 day, for naphthalene (0.1 mm) after 2 days and for phenanthrene (0.1 mm) after 15 days of cultivation. The conversion of anthracene (0.1 mm) and pyrene (0.1 mm) resulted in minimal residual concentrations, analogous to fluoranthene and pyrene of the anthracene oil (0.1%). Maximal total degradation for the tricyclic compounds dibenzofurane, fluorene, dibenzothiophene, phenanthrene and anthracene of the anthracene oil (0.1%) occurred after 5 days. In general, a significant metabolisation of the tetracyclic aromatic hydrocarbons fluoranthene and pyrene was observed after the degradation of phenol, naphthalene and most of the tricyclic compounds. Doubling the start concentrations of the polycyclic aromatic hydrocarbons effected higher degradation rates. Cell growth occurred simultaneously with the conversion of phenol, naphthalene and the tricyclic compounds. The immobilized cells showed stable growth and, compared to freely suspended cells, the same degradation sequence as well as an equivalent degradation potential — even in a model soil system. Correspondence to: I. Wiesel  相似文献   

12.
Physiological, biochemical and histological indices in Clarias gariepinus broodstock, and teratogenic indices in embryos exposed to sublethal concentrations of naphthalene, phenanthrene and pyrene were investigated in 2014 using a static-renewal bioassay protocol. Phenanthrene (1.41 mg l?1) was the most toxic, followed by pyrene (1.53 mg l?1) and naphthalene (7.21 mg l?1), based on 96 h LC50 values. Hepatosomatic indices were significantly higher in naphthalene- and pyrene-treated males compared with solvent controls, whereas fecundity in females was significantly lower by factors of 2.4 (naphthalene), 2.8 (phenanthrene) and 2.4 (pyrene), compared with controls. Catalase levels were lower in female phenanthrene-treated fish compared with controls. Histological alterations observed in PAH-treated fish include oedema, inflammatory cells, epithelial lifting and hyperplasia in the gills, vacuolation, haemosiderin pigments and sinusoidal congestion in the liver, and degenerated zona radiata in the ovary. Teratogenic effects were not observed, as evidenced by the lack of histological alterations in embryos spawned from pre-exposed broodstock. Sex-specific responses and the utility of biomarkers at cellular and individual levels of organisation are therefore demonstrated for holistic evaluations of polycyclic aromatic hydrocarbons in ecotoxicological studies.  相似文献   

13.
Giant freshwater prawns Macrobrachium rosenbergii (14-19 g) were challenged with Enterococcus (3 x 10(5) cfu prawn(-1)) previously incubated in TSB medium for 24 h, then placed in water having concentrations of dissolved oxygen (DO) at 7.75, 4.75, 2.75 and 1.75 mg l(-1). Onset of mortality occurred after 6 h exposure to 1.75 mg l(-1) DO, and after 12 h exposure to 2.75 mg 1(-1) DO. Cumulative mortality of prawns at 1.75 mg l(-1) DO was significantly higher than that at 4.75 and 2.75 mg l(-1) DO, and cumulative mortality of prawns at 4.75 and 2.75 mg l(-1) DO was significantly higher than that at 7.75 mg l(-1) DO after 96 h. The prawns (20-30 g) which had been placed in water for 0 to 120 h at 7.75, 4.75, 2.75 and 1.75 mg l(-1) DO were examined for the THC (total haemocyte counts), DHC (differential haemocyte counts), phenoloxidase activity, respiratory burst, percentage phagocytosis and clearance efficiency. No significant difference in semi-granular cells and granular cells of prawns was observed among four treatments. The prawns following 120 h exposure to 2.75 mg l(-1) DO decreased significantly the hyaline cells and THC by 39% and 36%, respectively. Phenoloxidase activity and respiratory burst decreased significantly by 33% and 11% when the prawns were exposed to 2.75 mg l(-1) DO after 24 h, respectively. Percentage phagocytosis and clearance efficiency to Enterococcus decreased significantly by 44% and 54% for the prawns following 12 h exposure to 2.75 mg l(-1) DO, respectively. It is concluded that DO as low as 2.75 mg l(-1) and 4.75 mg l(-1) causes depression in immune system of M. rosenbergii, and increases its susceptibility to Enterococcus infection.  相似文献   

14.
Phenanthrene is a three-ring polycyclic aromatic hydrocarbon and commonly found as a pollutant in various environments. Degradation of phenanthrene by white rot fungus Trametes versicolor 951022 and its laccase, isolated in Korea, was investigated. After 36 h of incubation, about 46% and 65% of 100 mg/l of phenanthrene added in shaken and static fungal cultures were removed, respectively. Phenanthrene degradation was maximal at pH 6 and the optimal temperature for phenanthrene removal was 30 degrees C. Although the removal percentage of phenanthrene was highest (76.7%) at 10 mg/l of phenanthrene concentration, the transformation rate was maximal (0.82 mg/h) at 100 mg/L of phenanthrene concentration in the fungal culture. When the purified laccase of T versicolor 951022 reacted with phenanthrene, phenanthrene was not transformed. The addition of redox mediator, 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) or 1-hydroxybenzotriazole (HBT) to the reaction mixture increased oxidation of phenanthrene by laccase about 40% and 30%, respectively.  相似文献   

15.
Litopenaeus vannamei juveniles (total length 22+/-2.4 mm) were exposed to different concentrations of ammonia-N (un-ionized plus ionized ammonia as nitrogen), using the static renewal method at different salinity levels of 15 per thousand, 25 per thousand and 35 per thousand at pH 8.05 and 23 degrees C. The 24, 48, 72, 96 h LC50 values of ammonia-N on L. vannamei juveniles were 59.72, 40.58, 32.15, 24.39 mg l(-1) at 15 per thousand; 66.38, 48.83, 43.17, 35.4 mg l(-1) at 25 per thousand; 68.75, 53.84, 44.93, 39.54 mg l(-1) at 35 per thousand, respectively. The 24, 48, 72, 96 h LC50 values of NH(3)-N (un-ionized ammonia as nitrogen) were 2.95, 2.00, 1.59, 1.20 mg l(-1) at 15 per thousand; 2.93, 2.16, 1.91, 1.57 mg l(-1) at 25 per thousand; 2.78, 2.18, 1.82, 1.60 mg l(-1) at 35 per thousand, respectively. As the salinity decreased from 35 per thousand to 15 per thousand, susceptibility of ammonia-N increased by 115%, 132%, 140% and 162% after 24, 48, 72 and 96 h exposure, respectively. The "safety level" for rearing L. vannamei juveniles was estimated to be 2.44, 3.55, 3.95 mg l(-1) for ammonia-N and 0.12, 0.16, 0.16 mg l(-1) for NH(3)-N in 15 per thousand, 25 per thousand and 35 per thousand, respectively.  相似文献   

16.
Microbial degradation of carbazole (CA), a model of hard-removal heterocyclic nitrogen compounds contained in petroleum oil, was examined using Sphingomonas sp. CDH-7 isolated from a soil sample by screening for CA-assimilating microorganisms. CDH-7 used CA as a sole source of carbon and nitrogen, and metabolized CA to ammonia via anthranilic acid as an intermediate product. When CDH-7 was cultivated in the medium containing CA at the concentration of 500 mg/l (2.99 mM), CA was completely degraded within 50 h. By the reaction with the resting cells of CDH-7, 500 mg/l of CA was completely degraded within 4 h, with 1.64 mM of ammonia accumulated in the reaction mixture. When CA was added at the concentration of 100 mg/l (0.599 mM) periodically to the reaction mixture ten times, 925 mg/l (5.54 mM) of CA was degraded within 48 h by the resting cells, and 4.50 mM of ammonia was accumulated in the reaction mixture with a 75.1% molar conversion yield based on total CA added. The resting cells could almost completely degrade CA in a two-liquid-phase system which consists of water and organic solvent, even in the presence of 20% (v/v) isooctane, n-hexane, cyclohexane, and kerosene as a model petroleum oil. In the presence of an organic solvent system such as 20% (v/v) pxylene, toluene, and heptanol, however, CA degradation yields decreased.  相似文献   

17.
Pseudomonas sp. strain HKT554 degrades naphtho[1,2- b]thiophene and two other isomers, naphtho[2,1- b]thiophene and naphtho[2,3- b]thiophene, by cometabolism, in the absence of any specific inducer, at similar degradation rates. A mutant of strain HKT554, deficient in dibenzothiophene degradation, was generated by using a recently developed transposition system. Sequence analysis of the mutant revealed that the knocked out gene was almost identical to naphthalene dioxygenase (EC 1.14.12.12). The mutant, HKT554M, degraded neither the naphthothiophene isomers nor dibenzothiophene, suggesting that the naphthalene dioxygenase is responsible for the initial catabolic reactions onto naphthothiophenes and dibenzothiophene.  相似文献   

18.
An algal-bacterial consortium formed by Chlorella sorokiniana and a phenanthrene-degrading Pseudomonas migulae strain was able to biodegrade 200-500 mg/l of phenanthrene dissolved in silicone oil or tetradecane under photosynthetic conditions and without any external supply of oxygen. Phenanthrene was only removed when provided in organic solvent, which confirms the potential of two-phase systems for toxicity reduction. Phenanthrene was degraded at highest rates when provided in silicone oil rather than in tetradecane since this solvent probably sequestered the PAH, reducing its mass transfer to the aqueous phase. The influence of phenanthrene concentration, amount of inoculum and light intensity on pollutant removal was also investigated and, under the best conditions, phenanthrene was degraded at 24.2 g m(-3).h(-1). In addition to being cost-effective and mitigating the release of greenhouse gases into the atmosphere, photosynthetic oxygenation was especially beneficial to the use of two-phase partitioning bioreactors since it prevented solvent emulsification and/or volatilization and evidence was found that the microalgae release biosurfactants that could further enhance phenanthrene degradation.  相似文献   

19.
In this study the degradation of hydrophobic petroleum model compounds (phenanthrene, pristane, octadecane and dibenzothiophene) added to a submersed hypersaline microbial mat was investigated. Montmorillonite with an artificially altered, hydrophobic surface was used as carrier material, forming an organo-clay complex (OCC) with the attached mixture of petroleum model compounds. 6 mg/cm2 OCC were applied to cyanobacterialmat pieces, containing 33.3 g/mg OCC of each compound. The degradationexperiment was performed under controlled laboratory conditions and accompanied bychemical analyses by GC/GC-MS, molecular analyses by PCR and DGGE as well asfunctional analyses by microsensor measurements of oxygen, photosynthesis, sulfide,pH and light. All applied model compounds were degraded, but residues were stillpresent after 18 weeks. The aromatic compounds phenanthrene (5.1 g/mg OCC)and dibenzothiophene (4.3 g/mg OCC) were preferentially degraded compared to the alkanes pristane (12.4 g/mg OCC) and n-octadecane (13.4 g/mg OCC). Metabolic changes during the degradation process could not be detected by microsensor measurements. The molecular population analyses did not reveal any significant community changes concomitant with the decrease of the petroleum model compounds. We conclude, that the pristine mats represent an intact, robust ecosystem in which the enzymatic requirements for the degradation of the applied pollutants exist. The slow degradation process did not affect the usual high internal turnover rates and did not favor a certain population in the community of the mats.  相似文献   

20.
A partitioning bioreactor, consisting of an aqueous phase containing Sphingomonas aromaticivorans and an immiscible organic phase (dodecane), loaded with naphthalene and phenanthrene, was operated at two scales, 5 l and 150 l. Complete degradation of 15 g and 300 g, respectively, of these polyaromatic hydrocarbon (PAH) mixtures was achieved in 21 h in both cases resulting in a volumetric PAH degradation rate of 238 mg l–1 h–1 based on reactor aqueous volumes.  相似文献   

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