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1.
Microbially produced lipopeptide have been isolated and studied for microbial enhanced oil recovery. About 60 gram positive bacteria isolated from soil contaminated with crude oil, near the crude oil storage tank in Tehran Refinery, Tehran, Iran. However, most of these studies have produced lipopeptide by one of the pure-culture microbes isolated in a laboratory. Among the isolates, heamolytic tests revealed two biosurfactant producers. The isolated strains were designated as C2, E1. By using morphological, biochemical and molecular biology tests (16 SrRNA), the strains identified as Bacillus licheniformis and Bacillus subtitlis, respectively. Emulsification activity and measurement of surface tension indicated that, the isolates were high producers of biosurfactant. The product of C2 and E1 is mainly lipopeptide. This product reduce surface tension from 65 to 30 mN/m. Emulsified activity of crude oil was 92% for C2 and 90 % in case of E1. This is the first report of indigenous Bacillus licheniformis and Bacillus subtilis from a soil contaminated with oil in an Iranian refinery with ability to produce biosurfactant.  相似文献   

2.
Crude oil and its derivatives because of different events and accidents may cause pollution to the environment. A biological treatment is a novel technique that uses microorganisms to remove or neutralize pollutants from a contaminated site. Oil-contaminated soils were sampled, after isolating of soil bacteria, using quantitative and qualitative screening, biosurfactant-producing bacteria were identified and environmental factors on the growth of bacteria and biosurfactant were investigated. In this study, the Bacillus subtilis was identified as the best biosurfactant-producing strain which has the ability to grow in environments with high salinity and temperature and pH > 5. The produced biosurfactant from B. subtilis is stable to changes in temperature and salt concentration and pH (in the range of 5–12).The B. subtilis also showed that they are able to biodegrade aliphatic alkanes. The B. subtilis has necessary potential for bioremediation of oil pollution in the environment.  相似文献   

3.
Halotolerant bacteria are regarded as effective oil-scavengers in the polluted saltern and seawater. In this regard, a halotolerant Planococcus was isolated from oil-contaminated area of Dezful north springs, Iran, due to its capacity in biosurfactant (BS) production. To facilitate hydrocarbons degradation, in the current study, the efficiency of BS production as function of growth rate of the halotolerant Planococcus was investigated in the vicinity of heavy crude oil by emulsification index (E24). Subsequently, the BS characterization was made by thin-layer chromatography (TLC), gas chromatography (GC) and infrared spectra analysis, and the stability was determined by E24 value measurement over a certain pH (5–9), temperature (20–100 °C) and salt concentration (0–10 % w/v) ranges. The BS production was found to be growth-associated. Detection of a unique band on TLC and GC chromatogram showed the extensive refining capacity of the BS purification, using the medium supernatant under acetone alkaline precipitation followed by oil dissolution from the sediment by carbon tetrachloride. Accordingly, it was clarified that the BS ultimately accumulated outside the cells. The glycolipid quality of the BS was further determined by the routine chemical characterization on TLC and by IR spectra analysis. Moreover, there was no protein detected by lowery total protein assay. Finally, the optimal temperature, pH and NaCl concentration to reach highest E24 values (85.7, 77.0, and 79.0 %) were found at respective 40 °C, pH = 9 and 0 % w/v. Our results revealed the practically potential of strain Dezful Isolate for BS large-scale production as environmentally friendly oil-eliminating agents.  相似文献   

4.
Isolation of hydrocarbonoclastic bacteria from bilge oil contaminated water   总被引：1，自引：1，他引：0  
Two bacterial strains, i.e. Pseudomonas mendocina and Ochrobactrum sp. were isolated from bilge oil contaminated water of Mormugao harbour, Goa, India and grown in a culture medium with hexadecane as the sole carbon source. Pseudomonas mendocina was used in further studies as it was the dominant strain. This strain effectively degraded tetradecane, hexadecane and octadecane leaving a residual concentration of about 73 %, 54 % and 40 % respectively in 120 h. Sequence analysis of the dominant bands from the denaturing gradient gel electrophoresis profiles revealed the differences between the genera of bilge oil contaminated sea water and its enrichment culture on hexadecane indicating a shift in community structure based on the type of substrate available. Pseudomonas mendocina amplified for the following catabolic genes namely C23O, nid and ndo. Based on the catabolic gene study the potential of the bacterial strain isolated, i.e. Pseudomonas mendocina seems to be interesting as it will be able to degrade polyaromatic hydrocarbons as well. Physicochemical properties of Pseudomonas mendocina indicates production of exopolysaccharides based on the value of its isoelectric point.  相似文献   

5.
This research work focuses on testing the bacterial strain Bacillus licheniformis for the bioremediation capacity of the crude oil. A biosurfactant and two different nanoparticles with different concentrations (0.05, 0.1, 0.2 g/100 ml) were applied separately to enhance the biodegradation process. The optimum biodegradation of crude oil was demonstrated at 60% of microcosms containing biosurfactant and nanoparticles after 7 days. The bacterial strain is highly potential to consume the total paraffins (iso- and n-paraffins) in crude oil samples. Accordingly, the best biodegradation of total paraffins was observed in microcosms containing (0.2 g) of Fe2O3, Zn5(OH)8Cl2 (nps) and biosurfactant separately. Additionally, the consumption of specific member rings of polyaromatics depends on the type and the concentration of nanoparticles. Thus, this bacterial strain was considered as a good candidate to be applied in the bioremediation process of petroleum-contaminated sites using biosurfactant and specific concentration of (Fe2O3 and Zn5OH8Cl2) nanoparticles.  相似文献   

6.
The lipA gene, encoding a solvent-tolerant extracellular lipase from Proteus sp. SW1, was displayed on the cell surface of Escherichia coli by fusing it to an antigen 43 anchoring motif. The display of LipA on the Escherichia coli cell surface was directly confirmed by immunofluorescence microscopy and flow cytometry. After 6 days of incubation in media containing 1 % used cooking oil, an Escherichia coli strain expressing surface displayed lipase was able to degrade 27 % of the oil. The biosurfactant, pseudopyronine B, was purified from culture supernatants of Pseudomonas sp. SL31. Its critical micelle concentration was determined to be 1400 mg/l, and the surfactant was stable within a temperature range from 0 to 120 °C and a pH range of 3–11. Pseudopyronine B-containing crude media extracts efficiently removed up to 51 % of the cadmium from contaminated water. We demonstrated the oil degradation ability of the mixed culture of four bacterial strains, namely the recombinant Escherichia coli expressing cell surface displayed lipase (pKKJlipA), His-tagged lipase (pETlipA), extracellular lipase-producing Proteus sp. SW1, and pseudopyronine B-producing Pseudomonas sp. SL31 by culturing in LB media containing 1 % oil. The consortium degraded 29 % of oil in one day and reached 84 % after 7 days.  相似文献   

7.
Production of microbial oil has attracted a great attention in recent years. The potential of lipid production by the yeast strains is the reason for using microorganisms for biodiesel production. Microbial lipid has high similarity to the oil obtained from plants and animals in type and composition. Production of oil from yeasts must be economical, so optimization of the cultivation condition to reach higher production must be done. Native oleaginous yeast, Cryptococcus albidus, was isolated from soil by the nitrogen-limited medium and screened by Nile red staining. Yarrowia lipolytica DSM 8218 was used for lipid production as a standard strain. C. albidus was an excellent oleaginous yeast, and the lipid quantity, dry biomass and lipid productivity of this strain were 11.81 g/l, 19.65 g/l and 60.1 %, respectively, in shaking flask cultivation at 150 rpm and 25 °C in nitrogen-limited medium containing per liter 75 g glucose, 1 g (NH4)2SO4, 1 g yeast extract, 3 g KH2PO4, 1.5 g MgSO4.7H2O, 0.15 g CaCl2, 0.06 g MnSO4.H2O, 0.02 g ZnSO4.7H2O and 0.15 g FeCl3.6H2O with pH adjusted to 6.5. Fourier transform infrared spectroscopy was used for analyzing and confirming the production of microbial oil in this study.  相似文献   

8.
An investigation was carried out to compare the ability of two bacteria Pseudomonas aeruginosa PSA5 and Rhodococcus sp. NJ2 isolated from petroleum sludge for degradation of benzo(a)pyrene [B(a)P], a HMW PAH compound in MSM. During 25 days of incubation, 50 ppm B(a)P was degraded by 88 and 47 % by P. aeruginosa PSA5 and Rhodococcus sp. NJ2, respectively. Besides, involvement of different catabolic enzymes, that is, salicylate hydroxylase, 2-carboxybenzaldehyde dehydrogenase, catechol 1,2-dioxygenase and catechol 2,3-dioxygenase, was also examined to identify their differential role in B(a)P degradation. Among these enzymes, the highest induction of 2-carboxybenzaldehyde dehydrogenase (773.5 nmol mg?1 protein) was recorded in P. aeruginosa PSA5, while salicylate hydroxylase was highly expressed (839.6 nmol mg?1 protein) in Rhodococcus sp. NJ2. Both the bacteria were found biosurfactant (glycolipid) producing, and role of biosurfactant in PAH degradation was also ascertained by reduced surface tension, higher emulsification index and increased cell surface hydrophobicity.  相似文献   

9.
Biodegradation of petroleum hydrocarbons as a decontamination mechanism is a relatively slow process. This study aimed to investigate the impact of a tailored consortium of bacteria with higher capacities in biosurfactant production and biodegradation on the acceleration of the biodecontamination process. To this end, 18 biosurfactant producing bacteria were isolated from the crude oil-contaminated soil samples of Isfahan refinery, and the activity of the produced biosurfactants was measured in terms of surface tension reduction and emulsification E24 test. Then, the isolates screened for the biodegradation of kerosene hydrocarbons and chemical structure of the purified biosurfactants produced by the most efficient isolates were partially characterized. Next, the isolates were sorted based on their surfactant activity and biodegradation efficiency, and the higher ranked bacteria thus selected were utilized to form an efficient consortium removing hydrocarbons from the oil-contaminated soil samples in a slurry phase system. The consortium consisted of Bacillus subtilis tb1 and Pseudomonas aeruginosa species having the highest biodegradation capabilities and surface activities. The results revealed that the hydrocarbon removal efficiency of the consortium was at least 25 % higher than single species, and the final removal efficiency for the consortium could be reached in a considerably shorter time.  相似文献   

10.
Experimental study of enhanced in-situ micro-ecological remediation of petroleum contaminated loess soil was carried out in Zhongyuan oil production areas, and the enhanced in-situ micro-ecological remediation technique includes optimistic in-situ microbial communities, physical chemistry methods, alfalfa planting and regulation of soil environmental elements. Experiments showed that the oil content in the contaminated soil with oil content about 2 898.25 mg/kg can be reduced about 98.61% after in-situ micro-ecological remediation for 99 days, which demonstrated the effectiveness of in-situ micro-ecological remediation methods for petroleum contaminated soil in central plains of China, and explored the practical and feasible application of these methods.  相似文献   

11.
The Lower Yangtze region is one of the important marine sedimentation areas of oil and gas distribution in southern China,for its favorable source rocks,reservoirs and covers.However,the intense tectonic movements and complex hydrocarbon generation process made it highly impossible to form large-sized oil and gas reservoirs.So it was divided to different hydrocarbon-bearing preservation units in oil-gas exploration.Recent study shows that the Permian and Lower Triassic source rocks in the Lower Yangtze region are complicated in lithology.The hydrocarbon generation potential of limestone there is low while argillaceous source rocks are overall of high abundance with excellent organic types,now in the process of hydrocarbon generation,so differences in high maturity influence the evaluation of organic matter abundance and type.Biomarker characteristics indicate a reductive environment.n-alkanes are marked by a single peak,with no odd-even predominance.The composition and distribution of the carbon numbers of n-alkanes,and the high abundance of long-chain tricyclic terpanes are indicative of marine sedi-mentation.The high contents of pregnane,homopregnane,rearranged hopane suggest that the source rocks are of high maturity.There is a good linear correlation between methylphenanthrene index and vitrinite reflectance.The correlation of oil-source rocks indicated that the oil of Well HT-3 may come from the Permian Longtan Formation in the Huangqiao area,the oil of Wells Rong-2 and Juping-1 came from the Lower Triassic Qinglong Formation in the Jurong area.The exploration here is promising in those different source rocks which all have great potential in hy-drocarbon generating,and oil and gas were produced in the late stage of hydrocarbon generation.  相似文献   

12.
Bioremediation strategies continue to be developed to mitigate the environmental impact of petroleum hydrocarbon contamination. This study investigated the ability of soil microbiota, adapted by prior exposure, to biodegrade petroleum. Soils from Barrow Is. (W. Australia), a class A nature reserve and home to Australia’s largest onshore oil field, were exposed to Barrow production oil (50 ml/kg soil) and incubated (25 °C) for successive phases of 61 and 100 days. Controls in which oil was not added at Phase I or II were concurrently studied and all treatments were amended with the same levels of additional nutrient and water to promote microbial activity. Prior exposure resulted in accelerated biodegradation of most, but not all, hydrocarbon constituents in the production oil. Molecular biodegradation parameters measured using gas chromatography–mass spectrometry (GC–MS) showed that several aromatic constituents were degraded more slowly with increased oil history. The unique structural response of the soil microbial community was reflected by the response of different phospholipid fatty acid (PLFA) sub-classes (e.g. branched saturated fatty acids of odd or even carbon number) measured using a ratio termed Barrow PLFA ratio (B-PLFAr). The corresponding values of a previously proposed hydrocarbon degrading alteration index showed a negative correlation with hydrocarbon exposure, highlighting the site specificity of PLFA-based ratios and microbial community dynamics. B-PLFAr values increased with each Phase I and II addition of production oil. The different hydrocarbon biodegradation rates and responses of PLFA subclasses to the Barrow production oil probably relate to the relative bioavailability of production oil hydrocarbons. These different effects suggest preferred structural and functional microbial responses to anticipated contaminants may potentially be engineered by controlled pre-exposure to the same or closely related substrates. The bioremediation of soils freshly contaminated with petroleum could benefit from the addition of exhaustively bioremediated soils rich in biota primed for the impacting hydrocarbons.  相似文献   

13.
Batch experiments were conducted to investigate As mobilization from mine tailings in the presence of a biosurfactant (JBR425, mixed rhamnolipids) and to evaluate the feasibility of using biosurfactant in remediating As contaminated mine tailings/soils. Introduction of the biosurfactant increased As mobilization greatly. When the mass ratio was 10 mg biosurfactant/g mine tailings at pH 11, As mobilization by the biosurfactant was greatest after 24 h, with a corresponding concentration ratio (the ratio of As mobilization by the biosurfactant to that by distilled water at same adjusted pH, wt/wt) of 21.6. Selective sequential extraction indicated that As was easily mobilized from the weakly bound and relatively more mobile fractions by washing with the biosurfactant. A mobilization isotherm was developed to predict As mobilization from the mine tailings in the presence of biosurfactant. It was shown that biosurfactant sorption to the mine tailings is essential for As mobilization. Arsenic mobilization was found to be positively correlated with the mobilization of Fe and other metals (i.e., Cu, Pb and Zn), which might further enhance As mobilization by helping incorporate it into soluble complexes or micelles. Capillary electrophoresis analyses indicated that As redox or methylation reactions had insignificant effect on As mobilization. Biosurfactants might be used potentially to remove bulk As from mine tailings or contaminated soils under alkaline conditions.  相似文献   

14.
The detection of microorganisms with potential for biodeterioration and biodegradation in petroleum fields is of great relevance, since these organisms may be related to a decrease in petroleum quality in the reservoirs or damage in the production facilities. In this sense, petroleum formation water and oil samples were collected from the Campos Basin, Brazil, with the aim of isolating microorganisms and evaluating their ability to degrade distinct classes of hydrocarbon biomarkers (9,10-dihydrophenanthrene, phytane, nonadecanoic acid and 5α-cholestane). Twenty eight bacterial isolates were recovered and identified by sequencing their 16S rRNA genes. Biodegradation assays revealed that bacterial metabolism of hydrocarbons occurred through reactions based on oxidation, carbon–carbon bond cleavage and generation of new bonds or by the physical incorporation of hydrocarbons into microbial cell walls. Based on the biodegradation results, selective PCR-based systems were developed for direct detection in petroleum samples of bacterial groups of interest, namely Bacillus spp., Micrococcus spp., Achromobacter xylosoxidans, Dietzia spp. and Bacillus pumilus. Primer sets targeting 16S rRNA genes were designed and their specificity was confirmed in silico (i.e. computational analysis) and in PCR reactions using DNA from reference strains as positive and negative controls. Total DNA from oil was purified and the amplification tests revealed the presence of the target bacteria in the samples, unraveling a significant potential for petroleum deterioration in the reservoirs sampled, once proper conditions are present for hydrocarbon degradation. The application of molecular methods for rapid detection of specific microorganisms in environmental samples would be valuable as a supporting tool for the evaluation of oil quality in production reservoirs.  相似文献   

15.
微生物修复石油污染地下水的实验研究   总被引：6，自引：0，他引：6  
为修复陕北黄土区石油污染地下水,采用优化土著微生物菌群的生物技术,进行了地下水中石油的降解与修复实验研究。在实验装置内加入了1.5%的优化菌群制剂,优化出的菌群初步鉴定主要有:假单胞菌属、微球菌属、放线菌属、真菌类的青霉属和曲霉属等。实验结果显示,在实验装置中人为添加石油含量为182.5 mg/l、862.5 mg/l、1695.0 mg/l时,经过28~37 d的微生物修复,地下水中石油的降解率为27.47%~92.46%,而无菌对照中的石油含量变化在5%以内,说明在无菌条件下地下水中石油降解缓慢。该实验过程验证了微生物修复技术在地下水石油污染修复中的有效性,探索了应用的可行性。  相似文献   

16.
Biodegradation, one of the most important weathering processes, alters the composition of spilled oil, making it difficult to identify the source of the release and to monitor its fate in the environment. A laboratory experiment was conducted to simulate oil spill weathering process of microbial degradation to investigate compositional changes in a range of source- and weathering-dependent molecular parameters in oil residues, and the conventional diagnostic ratios for oil spill identification were also evaluated. The conventional diagnostic ratios of n-alkane displayed obvious changes after biodegradation, especially for Pr/n-C17 and Ph/n-C18 with relative standard deviation more than 118.84 %, which suggests they are invalid for oil source identification of the middle-serious spill. Many polycyclic aromatic hydrocarbons (PAHs) are more resistant to biodegradation process than their saturated hydrocarbon counterparts, thus making PAHs to be one of the most valuable fingerprinting classes of hydrocarbons for oil identification. Biomarker ratios of hopanes and steranes were also useful for source identification even after moderate biodegradation, and the diagnostic ratios from them could be used in tracking origin and sources of hydrocarbon pollution. Finally, the carbon isotopic type curve may provide another diagnostic means for correlation and differentiation of spilled oils, and be particularly valuable for lighter refined products or severely biodegraded oils, the source of which may be difficult to identify by routine biomarker techniques.  相似文献   

17.
The water and sediments of Lake Coeur d’Alene in northern Idaho (USA) have been impacted by decades of mining operations within the Coeur d’Alene mining district. Using a multivariate statistical approach, correlations were explored between the microbial community (via 16S rDNA microarray) in sediment cores and operationally defined heavy metal phases (via continuous sequential extractions). Candidate phyla NC10, OP8 and LD1PA were only detected in metal contaminated cores and diversity doubled among Natronoanaerobium in metal contaminated cores compared to the uncontaminated control site. This may suggest some increased fitness of these phyla in contaminated sediments. In contrast, diversity within the phyla Aquificae, Coprothermobacteria, and Synergistes was at least double in the uncontaminated control site. In linear models composed of two geochemical variables from the presumed sulfate reducing lineages detected in this study, orders Desulfobacterales, Desulfuromonadales, Desulfotomaculum, and Syntrophobacterales were highly correlated with Pb (positive influence) and Zn (negative influence) in the operationally defined residual fraction, and most taxa within orders from Desulfovibrionales. Bdellovibrionales highly correlated with Pb in the exchangeable/carbonate (negative influence) and oxyhydroxide (positive influence) phases. Diversity within families from metal reducing bacterial lineages Shewanellaceae, Geobacteraceae, and Rhodocyclaceae showed high correlation with Pb in the exchangeable/carbonate (negative influence) and oxyhydroxide (positive influence) phases. To our knowledge, this is the first time these techniques have been used in combination to describe a contaminated system. Resulting correlations suggest the diversity of the microbial community was influenced primarily by partitioning of heavy metals into exchangeable Pb over other Pb phases and, to a lesser extent, residual Pb to residual Zn phase partitioning.  相似文献   

18.
Carbon isotopic compositions were determined by GC–IRMS for individual n-alkanes in crude oils and the free, adsorbed and inclusion oils recovered by sequential extraction from reservoir rocks in the Tazhong Uplift and Tahe oilfield in the Tabei Uplift of Tarim Basin as well as extracts of the Cambrian–Ordovician source rocks in the basin. The variations of the δ13C values of individual n-alkanes among the 15 oils from the Tazhong Uplift and among the 15 oils from the Triassic and Carboniferous sandstone reservoirs and the 21 oils from the Ordovician carbonate reservoirs in the Tahe oilfield demonstrate that these marine oils are derived from two end member source rocks. The major proportion of these marine oils is derived from the type A source rocks with low δ13C values while a minor proportion is derived from the type B source rocks with high δ13C values. Type A source rocks are within either the Cambrian–Lower Ordovician or the Middle–Upper Ordovician strata (not drilled so far) while type B source rocks are within the Cambrian–Lower Ordovician strata, as found in boreholes TD2 and Fang 1. In addition, the three oils from the Cretaceous sandstone reservoirs in the Tahe oilfield with exceptionally high Pr/Ph ratio and δ13C values of individual n-alkanes are derived, or mainly derived, from the Triassic–Jurassic terrigenous source rocks located in Quka Depression.The difference of the δ13C values of individual n-alkanes among the free, adsorbed and inclusion oils in the reservoir rocks and corresponding crude oils reflects source variation during the reservoir filling process. In general, the initial oil charge is derived from the type B source rocks with high δ13C values while the later oil charge is derived from the type A source rocks with low δ13C values.The δ13C values of individual n-alkanes do not simply correlate with the biomarker parameters for the marine oils in the Tazhong Uplift and Tahe oilfield, suggesting that molecular parameters alone are not adequate for reliable oil-source correlation for high maturity oils with complex mixing.  相似文献   

19.
环境温度影响石油黏滞性,改变石油污染土的力学特性。借助无侧限抗压强度试验,研究不同温度条件下石油污染滨海盐渍土的强度及变形特性。结果证实:温度对石油污染盐渍土力学性质影响显著,温度越高,污染土抗压强度越低,20℃时,含油率15%污染土的抗压强度较未污染土下降近70%;石油污染滨海盐渍土的抗压强度随含油率的增加先增大后减小,环境温度与污染土强度峰值点处的含油率反相关,10℃、20℃、30℃条件下,强度峰值点处的含油率分别为15%、10%和5%;石油污染盐渍土表现为应变软化型破坏,含油率及环境温度影响石油污染盐渍土的抗变形能力,土样的破坏面积和裂缝宽度随含油率增加及温度升高而增大。  相似文献   

20.
The efficiency of inorganic fertilizers as stimulating agents for the bioremediation of oil-polluted environments can be increased with the addition of selected biostimulating compounds. In this study, the efficacy of different biostimulation treatments in the remediation of diesel-polluted soil in purpose-built microcosms has been evaluated. The treatments involved combinations of inorganic fertilizer with (a) Ivey surfactant, (b) Biorem organic fertilizer and (c) ethanol. Microbial activity was evaluated by monitoring the growth of heterotrophic and degrading bacteria and their dehydrogenase activity and carbon dioxide production. Hydrocarbon degradation was monitored by gas chromatography/mass spectrometry. The results showed that all treatments enhanced microbial activity in comparison with natural attenuation and also that the combined treatments generally enhanced hydrocarbon biodegradation in comparison to both natural attenuation and the single inorganic fertilizer treatment. The inorganic fertilizer plus Ivey? surfactant was the most efficient treatment in terms of Total Petroleum Hydrocarbon and light and heavy n-alkanes, showing an index of degradation of 1.4 and 1.3, respectively. Furthermore, biodegradation of heavy and branched n-alkanes was higher in microcosms treated with inorganic fertilizer plus ethanol (Index of degradation values of 1.6 and 1.5, respectively) indicating that combined treatments can be very effective in restoration of contaminated soil.  相似文献   

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