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1.
Armillaria sp. F022, a white-rot fungus isolated from decayed wood in tropical rain forest was used to biodegrade anthracene in cultured medium. The percentage of anthracene removal by Armillaria sp. F022 reached 13 % after 7 days and at the end of the experiment, anthracene removal level was at 87 %. The anthracene removal through sorption and transformation was investigated. 69 % of eliminated anthracene was transformed by Armillaria sp. F022 to form other organic structure, while only 18 % was absorbed in the mycelia. In the kinetic experiment, anthracene dissipation will not stop even though the biomass had stopped growing. Anthracene removal by Armillaria sp. F022 was correlated with protein concentration (whole biomass) in the culture. The production of enzyme was affected by biomass production. Anthracene was transformed to two stable metabolic products. The metabolites were extracted in ethyl-acetate, isolated by column chromatography, and then identified using gas chromatography–mass spectrometry (GC–MS).  相似文献   

2.
The effects of structure and concentration of surfactants on the biodegradation of fluoranthene, a three rings polycyclic aromatic hydrocarbon in the aqueous phase, as well as their effects on the biodegradation and enzyme activity were investigated. The toxicity ranking of studied surfactants is: non-ionic Tween 80 <anionic sodium dodecyl sulfate <cationic Tetradecyltrimethylammonium bromide. The maximum growth of Armillaria sp. F022 (>4,500 mg/L) was showed by Tween 80 (10 mg/L) culture, manifesting that the non-ionic surfactant present in the culture were beneficial to the fungal growth. Laccase showed the highest enzymes activity in all surfactants culture. Non-ionic Tween 80 showed a significant result for laccase activity (1,902 U/L) in the Armillaria sp. F022 culture. The increased enzymes cumulative activity may stem directly from the rising fluoranthene biodegradability as addition of appropriate surfactants. The biotransformation of fluoranthene was greatly improved by Tween 80, and totally fluoranthene degradation was obtained as Tween 80 was 10 mg/L. Two fluoranthene metabolites were isolated from the culture medium and analyzed by a thin layer chromatography, UV visible spectrometer and gas chromatography–mass spectrometry (GC–MS). The oxidation of fluoranthene is initiated by oxygenation at the C-2,3 positions resulting 9-fluorenone. At the end of experiment, one metabolite was detected in the culture extract and identified as phthalic acid. Evidently, Armillaria sp. F022 seems efficient, high effective and deserves further application on the enhanced bioremediation technologies for the treatment of fluoranthene-contaminated soil.  相似文献   

3.
Characterization of anthracene metabolites produced by Armillaria sp. F022 was performed in the enzymatic system. The fungal culture was conducted in 100-mL Erlenmeyer flask containing mineral salt broth medium (20 mL) and incubated at 120 rpm for 5–30 days. The culture broth was then centrifuged at 10,000 rpm for 45 min to obtain the extract. Additionally, the effect of glucose consumption, laccase activity, and biomass production in degradation of anthracene were also investigated. Approximately, 92 % of the initial concentration of anthracene was degraded within 30 days of incubation. Dynamic pattern of the biomass production was affected the laccase activity during the experiment. The biomass of the fungus increased with the increasing of laccase activity. The isolation and characterization of four metabolites indicated that the structure of anthracene was transformed by Armillaria sp. F022 in two routes. First, anthracene was oxidized to form anthraquinone, benzoic acid, and second, converted into other products, 2-hydroxy-3-naphthoic acid and coumarin. Gas chromatography–mass spectrometry analysis also revealed that the molecular structure of anthracene was transformed by the action of the enzyme, generating a series of intermediate compounds such as anthraquinone by ring-cleavage reactions. The ligninolytic enzymes expecially free extracellular laccase played an important role in the transformation of anthracene during degradation period.  相似文献   

4.
A diverse surfactant, including the nonionic Tween 80 and Brij 30, the anionic sodium dodecyl sulphate, the cationic surfactant Tetradecyltrimethylammonium bromide, and biosurfactant Rhamnolipid were investigated under fluorine-enriched medium by Armilaria sp. F022. The cultures were performed at 25 °C in malt extract medium containing 1 % of surfactant and 5 mg/L of fluorene. The results showed among the tested surfactants, Tween-80 harvested the highest cell density and obtained the maximum specific growth rate. This due Tween-80 provide a suitable carbon source for fungi. Fluorane was also successfully eliminated (>95 %) from the cultures within 30 days in all flasks. During the experiment, laccase production was the highest among other enzymes and Armillaria sp. F022-enriched culture containing Non-ionic Tween 80 showed a significant result for laccase activity (1,945 U/L). The increased enzyme activity was resulted by the increased biodegradation activity as results of the addition of suitable surfactants. The biotransformation of fluorene was accelerated by Tween 80 at the concentration level of 10 mg/L. Fluorene was initially oxidized at C-2,3 positions resulting 9-fluorenone. Through oxidative decarboxylation, 9-fluorenone subjected to meta-cleavage to form salicylic acid. One metabolite detected in the end of experiment, was identified as catechol. Armillaria sp. F022 evidently posses efficient, high effective degrader and potential for further application on the enhanced bioremediation technologies for treating fluorene-contaminated soil.  相似文献   

5.
Pleurotus pulmonarius F043, a fungus collected from tropical rain forest, was used to degrade pyrene, a four-rings polycyclic aromatic hydrocarbons (PAHs), in a mineral medium broth. A maximum degradation rate of pyrene (90 %) was occurred at pH 3 and the lowest degradation rate was found in the culture at pH 10 (2 %). More than 90 % pyrene degradation was achieved at pH ranged from 3 to 5, whereas the degradation rate significantly declined when the pH was >5. The degradation of pyrene increased from 2 to 96 % when the temperature rose from 4 to 25 °C. When the temperature was increased to 60 °C resulting the lowest degradation rate into 7 %. Among the agitation rates tested, 120 rpm was the best with 95 % degradation, followed by 100 rpm (90 %). The optimum agitation range for pyrene degradation by P. pulmonarius F043 was 100–120 rpm. Among all the concentrations tested, 0.5 % Tween 80 was the best with 98 % degradation, followed by 1 % Tween 80 (90 %). The optimum concentration of Tween 80 for pyrene degradation by P. pulmonarius F043 was 0.5–1 %. The degradation rate decreased, while the concentration of Tween 80 was increased. The metabolic product was found during degradation process through the identification of gentisic acid by TLC, UV-Spectrophotometer, and GC–MS.  相似文献   

6.
For the first time the dependence of completeness of pyrene degradation by the white-rot fungus Pleurotus ostreatus D1 on cultivation conditions was found. In Kirk’s medium about 65.6 ± 0.9% of the initial pyrene was metabolized after 3 weeks, with pyrene-4,5-dihydrodiol accumulating. This process was accompanied by laccase production only. In basidiomycetes rich medium, P. ostreatus D1 metabolized up to 89.8 ± 2.3% of pyrene within 3 weeks without pyrene-4,5-dihydrodiol accumulation throughout the time of cultivation. Phenanthrene and phthalic acid were identified as the metabolites produced from pyrene degradation under these conditions. Accumulation of phenanthrene with its subsequent disappearance was observed. One more metabolite probably was the product of phenanthrene degradation. Pyrene metabolism in basidiomycetes rich medium was accompanied first by laccase and tyrosinase production and later by versatile peroxidase production. The cell-associated activities of laccase, tyrosinase, and versatile peroxidase were found. The data obtained indicate that both enzymes (laccase and versatile peroxidase) are necessary for complete degradation of pyrene. Furthermore, both cell-associated and extracellular laccases can catalyse the first stages of pyrene degradation, and versatile peroxidase can be necessary for oxidation of the resulting metabolites.  相似文献   

7.
The white-rot shiitake mushroom, Lentinula edodes, was used to degrade an environmentally hazardous compound, 2,4-dichlorophenol (DCP), using vanillin as an activator. Vanillin increased the mycelial growth from 74 to 118 mg/150 ml culture and accelerated laccase and Mn-peroxidase production from the maximum on days 24–28 without vanillin to days 10–14. It eliminated 92 % of 100 mM DCP with 50 mg vanillin/l compared with only 15 % without vanillin. GC–MS revealed that a diaryl ether dimer of DCP was formed in the culture without vanillin, whereas dimer formation was diminished with vanillin addition. This indicates that vanillin enhances the degradation of DCP and disrupts the formation of the toxic dimer. Therefore, lignin-derived phenol such as vanillin can be used as natural and eco-friendly activators to control white-rot mushrooms, thereby facilitating the effective degradation of environmentally hazardous compounds.  相似文献   

8.
Forty-two white-rot fungi in submerged cultures were tested to determine their dye decolorization capacity and the optimal conditions for the decolorization process. Trametes pubescens Cui 7571 was found to be the most effective strain in terms of decolorization performance on the azo dye Congo Red, and it exhibited excellent reusability as well as persistence in sequential decolorization experiments. Optimization of the decoloration process was also conducted to evaluate the effects of a number of chemical compounds, metal salts, inducers, and mediators on the dye decolorization rate. On the seventh day, a highest dye removal of 98.83 % was observed with addition of copper at 2.5 mmol L?1, Tween 80 at 1.0 % (v/v), and ferulic acid at 0.50 μmol L?1, respectively. The adsorption of mycelia to dyes was not a significant contributor to dye removal, and decolorization by the functional fungus T. pubescens depended on biodegradation by enzymes, as evidenced by the results of the moist heat sterilization treatment (121°C for 20 min), induction of extracellular enzymes, and scanning electron microscopy. Four dye degradation metabolites, i.e., naphthalene amine, biphenyl amine, biphenyl ,and naphthalene diazonium, were identified by Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry. The phytotoxicity tests indicated that degraded metabolites had almost a negligible effect on the plant seeds as compared to that of dye, which is indicative of the less toxic nature of the metabolites. Our results suggest that white-rot fungus T. pubescens could be developed into a novel azo dye bioremediation strategy.  相似文献   

9.
The white-rot fungus Pleurotus eryngii F032 showed the capability to degrade a three fused-ring aromatic hydrocarbons fluorene. The elimination of fluorene through sorption was also investigated. Enzyme production is accompanied by an increase in biomass of P. eryngii F032 during degradation process. The fungus totally degraded fluorine within 23 d at 10-mg l−1 solution. Fluorene degradation was affected with initial fluorene concentrations. The highest enzyme activity was shown by laccase in the 10-mg l−1 culture after 30 d of incubation (1620 U l−1). Few activities of enzymes were observed in the fungal cell at the varying concentration of fluorene. Three metabolic were detected and separated in ethylacetate extract, after isolated by column chromatography. The metabolites, 9-fluorenone, phthalic acid, and benzoic acid were identified using UV–vis spectrophotometer and gas chromatography–mass spectrometry (GC–MS). The results show the presence of a complex mechanism for the regulation of fluorene-degrading enzymes.  相似文献   

10.
A fusant strain F14 with high biodegradation capability of phenanthrene was obtained by protoplast fusion between Sphingomonas sp. GY2B (GenBank DQ139343) and Pseudomonas sp. GP3A (GenBank EU233280). F14 was screened and identified from 39 random fusants by antibiotic tests, scanning electron microscope (SEM) and randomly amplified polymorphic DNA (RAPD). The result of SEM analysis demonstrated that the cell shape of fusant F14 different from parental strains. RAPD analysis of 5 primers generated a total of 70 bands. The genetic similarity indices between F14 and parental strains GY2B and GP3A were 27.9 and 34.6 %, respectively. F14 could rapidly degrade phenanthrene within 24 h, and the degradation efficiency was much better than GY2B and GP3A. GC–MS analysis of metabolites of phenanthrene degradation indicated F14 had a different degradation pathway from GY2B. Furthermore, the fusant strain F14 had a wider adaptation of temperatures (25–36 °C) and pH values (6.5–9.0) than GY2B. The present study indicated that fusant strain F14 could be an effective and environment-friendly bacterial strain for PAHs bioremediation.  相似文献   

11.
As a discarded lignocellulosic biomass, chestnut shell is of great potential economic value, thus a sustainable strategy is needed and valuable for utilization of this resource. Herein, the feasibility of biological processes of chestnut shell with Dichomitus squalens, Phlebia radiata and their co-cultivation for lignin-modifying enzymes (LMEs) production and biodegradation of this lignocellulosic biomass was investigated under submerged cultivation. The treatment with D. squalens alone at 12 days gained the highest laccase activity (9.42 ± 0.73 U mg?1). Combined with the data of laccase and manganese peroxidase, oxalate and H2O2 were found to participate in chestnut shell degradation, accompanied by a rapid consumption of reducing sugar. Furthermore, specific surface area of chestnut shell was increased by 77.6–114.1 % with the selected fungi, and total pore volume was improved by 90.2 % with D. squalens. Meanwhile, the surface morphology was observably modified by this fungus. Overall, D. squalens was considered as a suitable fungus for degradation of chestnut shell and laccase production. The presence of LMEs, H2O2 and oxalate provided more understanding for decomposition of chestnut shell by the white-rot fungi.  相似文献   

12.

Background and aims

The selective inoculation of specific hydrocarbon-degrading microbes into the plant rhizosphere offers a useful means for remediating hydrocarbon-contaminated soils. The effect of inoculating a seed-borne filamentous fungus (Lewia sp.) on hydrocarbon removal by Festuca arundinacea and its growth was studied on perlite (model soil) and soil, both spiked with hydrocarbons.

Methods

A hydrocarbon mixture (1,500 mg kg?1) of two polycyclic aromatic hydrocarbons (PAH), phenanthrene and pyrene, blended with hexadecane (1.0:0.5:0.5 weight) was used. Greenhouse experiments were carried out for 45 days. Inoculated and non-inoculated plants were grown in dark cylindrical glass pots containing perlite or soil.

Results

Inoculation with Lewia sp. stimulated (100 %) root growth in spiked perlite. Inoculated plants showed higher phenanthrene removal (100 %) compared to non-inoculated plants in perlite and soil. Pyrene removal by inoculated plants was 37-fold higher than that by non-inoculated plants in perlite; in soil, pyrene removal by inoculated plants (97.9 %) differed significantly from that of non-inoculated plants (91.4 %). Accumulation of pyrene in roots (530.9 mg kg?1 of dry roots) was promoted in perlite.

Conclusions

Our results demonstrate that Lewia sp. (endophytic fungus) improved the efficiency of PAH removal by F. arundinacea, on both perlite and soil, stimulating pyrene accumulation in roots.  相似文献   

13.
Agrocybe sp. CU-43, a white-rot fungus isolated from Thailand, showed a high potential for degrading both low- and high-molecular weight polycyclic aromatic hydrocarbons. At 100 ppm fluorene was degraded by 99% within six days while at the same concentration 99 and 92% degradation of phenanthrene and anthracene, respectively, occurred in 21 days, and fluoranthene and pyrene were reduced by 80 and 75%, respectively, in 30 days. In a soil model, Agrocybe sp. CU-43 completely degraded 250 ppm fluorene at room temperature within four weeks. Laccase and manganese peroxidase activities, but not lignin peroxidase activity, were detected during the biodegradation of fluorene. Two of the metabolites from fluorene degradation by the fungus were identified via reversed-phase HPLC as 9-fluorenol and 9-fluorenone, the less toxic intermediates of fluorene. However, 9-fluorenol is not an end product for the degradation. These results suggest that fluorene degradation by Agrocybe sp. CU-43 may take place via the same pathway(s) employed by other ligninolytic and non-ligninolytic fungi. This is the first report of fluorene biodegradation by a fungus belonging to the genus Agrocybe.  相似文献   

14.
Pyrene degradation is known in bacteria. In this study, Mycobacterium sp. strain KMS was used to study the metabolites produced during, and enzymes involved in, pyrene degradation. Several key metabolites, including pyrene-4,5-dione, cis-4,5-pyrene-dihydrodiol, phenanthrene-4,5-dicarboxylic acid, and 4-phenanthroic acid, were identified during pyrene degradation. Pyrene-4,5-dione, which accumulates as an end product in some gram-negative bacterial cultures, was further utilized and degraded by Mycobacterium sp. strain KMS. Enzymes involved in pyrene degradation by Mycobacterium sp. strain KMS were studied, using 2-D gel electrophoresis. The first protein in the catabolic pathway, aromatic-ring-hydroxylating dioxygenase, which oxidizes pyrene to cis-4,5-pyrene-dihydrodiol, was induced with the addition of pyrene and pyrene-4,5-dione to the cultures. The subcomponents of dioxygenase, including the alpha and beta subunits, 4Fe-4S ferredoxin, and the Rieske (2Fe-2S) region, were all induced. Other proteins responsible for further pyrene degradation, such as dihydrodiol dehydrogenase, oxidoreductase, and epoxide hydrolase, were also found to be significantly induced by the presence of pyrene and pyrene-4,5-dione. Several nonpathway-related proteins, including sterol-binding protein and cytochrome P450, were induced. A pyrene degradation pathway for Mycobacterium sp. strain KMS was proposed and confirmed by proteomic study by identifying almost all the enzymes required during the initial steps of pyrene degradation.  相似文献   

15.
A new yeast strain was isolated from sugarcane cultivation field which was able to utilize lindane as sole carbon source for growth in mineral medium. The yeast was identified and named as Candida sp. VITJzN04 based on a polyphasic approach using morphological, biochemical and 18S rDNA, D1/D2 and ITS sequence analysis. The isolated yeast strain efficiently degraded 600 mg L?1 of lindane within 6 days in mineral medium under the optimal conditions (pH 7; temperature 30 °C and inoculum dosage 0.06 g L?1) with the least half-life of 1.17 days and degradation constant of 0.588 per day. Lindane degradation was tested with various kinetic models and results revealed that the reaction could be described best by first-order and pseudo first-order models. In addition, involvement of the enzymes viz. dechlorinase, dehalogenase, dichlorohydroquinone reductive dechlorinase, lignin peroxidase and manganese peroxidase was noted during lindane degradation. Addition of H2O2 in the mineral medium showed 32 % enhancement of lindane degradation within 3 days. Based on the metabolites identified by GC–MS and FTIR analysis, sequential process of lindane degradation by Candida VITJzN04 was proposed. To the best of our knowledge, this is the first report of isolation and characterization of lindane-degrading Candida sp. and elucidation of enzyme systems during the degradation process.  相似文献   

16.
Pyrene as high molecular weight polycyclic aromatic hydrocarbons (PAHs) is found in most terrestrial environment. However, the biodegradation of pyrene has been limited due to its bioavailability and toxicity. So it is vital to study degradation-capable microorganism and suitable co-substrate. In this study, the indigenous white-rot fungus Pseudotrametes gibbosa isolated from ChangBai Mountain located in northeast China was used to degrade pyrene, and 6 co-substrates were selected as co-metabolic carbon and energy sources. The results showed that P. gibbosa was able to utilize pyrene as sole carbon and energy source. The degradation efficiency achieved 28.33% within 18 days. Meanwhile, co-substrate wheat bran extract could stimulate laccase (Lac) production significantly by P. gibbosa, compared with other co-substrates and control (without co-substrate). In the presence of co-substrates, the biodegradation efficiency of pyrene ranged from 34.23% to 50.64% which was enhanced due to co-metabolism except for salicylic acid (25.91%) and phthalic acid (21.64%).  相似文献   

17.
Many reports describe the decolourization of dyes by fungal enzymes. However, these enzymes do not contribute to dye mineralization but only to its biotransformation into less coloured or colourless molecules persisting in solution. Therefore, it is essential to analyse the identity of the metabolites produced during enzymatic treatments and its biodegradation into an appropriate system. The present work examines the decolourization/detoxification of a simulated effluent (containing Acid Blue 74) by fungal enzymes and proposes a secondary treatment using an anaerobic system to improve the enzymatic decolourization through the complete mineralization of the dye. Ligninolytic enzymes were produced by solid culture using the thermo-tolerant fungus Fomes sp. EUM1. The enzymes produced showed a high rate of decolourization (>95 % in 5 h) and were stable at elevated temperature (40 °C) and ionic strength (NaCl, 50 mM). Isatin-5-sulphonic acid was identified via 1H-NMR as oxidation product; tests using Daphnia magna revealed the non-toxic nature of this compound. To improve the enzymatic degradation and avoid coupling reactions between the oxidation products, the effluent was subjected to an anaerobic (methanogenic) treatment, which achieved high mineralization efficiencies (>85 %). To confirm the mineralization of isatin-5-sulphonic acid, a specific degradation study, which has not been reported before, with this single compound was conducted under the same conditions; the results showed high removal efficiencies (86 %) with methane production as evidence of mineralization. These results showed the applicability of an anaerobic methanogenic system to improve the enzymatic decolourization/detoxification of Acid Blue 74 and achieve its complete mineralization.  相似文献   

18.
Kraft lignin (KL) is the major pollutant in black liquor. The bacterial strain Pandoraea sp. B-6 was able to degrade KL without any co-substrate under high alkaline conditions. At least 38.2 % of chemical oxygen demand and 41.6 % of color were removed in 7 days at concentrations from 1 to 6 g L?1. The optimum pH for KL degradation was 10 and the optimum temperature was 30 °C. The greatest activities of 2,249.2 U L?1 for manganese peroxidase and 1,120.6 U L?1 for laccase were detected on the third and fifth day at pH 10, respectively. Many small molecules, such as cinnamic acid, ferulic acid, 2-hydroxy benzyl alcohol, and vanillyl methyl ketone, were formed during the period of KL degradation based on GC–MS analysis. These results indicate that this strain has great potential for biotreatment of black liquor.  相似文献   

19.
Polyporus sp. S133 decolorized the Amaranth in 72 h (30 mg L?1) under static and shaking conditions. Liquid medium containing glucose has shown the highest decolorization of Amaranth by Polyporus sp. S133. When the effect of increasing inoculum concentration on decolorization of Amaranth was studied, maximum decolorization was observed with 15 % inoculum concentration. Significant increase in the enzyme production of laccase (102.2 U L?1) was observed over the period of Amaranth decolorization compared to lignin peroxidase and manganese peroxidase. Germination rate of Sorghum vulgare and Triticum aestivum was less with Amaranth treatment as compared to metabolites obtained after its decolorization. Based on the metabolites detected by GC–MS, it was proposed that Amaranth was bio-transformed into two intermediates, 1-hydroxy-2-naphthoic acid and 1,4-naphthaquinone. Overall findings suggested the ability of Polyporus sp. S133 for the decolorization of azo dye and ensured the ecofriendly degradation of Amaranth.  相似文献   

20.
In this study, nitriles were used as sole sources of nitrogen in the enrichments to isolate nitrile-converting microorganisms. A novel fungus named ZJB-09150 possessing nitrile-converting enzymes was obtained with 3-cyanopyridine as sole source of nitrogen, which was identified by morphology, biology and 18S rDNA gene sequence as Fusarium proliferatum. It was found that F. proliferatum had ability to convert nitriles to corresponding acids or amides and showed wide substrate specificity to aliphatic nitriles, aromatic nitriles and ortho-substituted heterocyclic nitriles. The nitrile converting enzymes including nitrilase and nitrile hydratase in ZJB-09150 were induced by ε-caprolactam. Nitrilase obtained in this study showed high activity toward 3-cyanopyridine. It was active within pH 3.0–12.0 and temperature ranging from 25 to 65 °C with optimal at pH 9.0 and temperature 50–55 °C. The enzyme was thermostable and its half-life was 12.5 and 6 h at 45 and 55 °C, respectively. Under optimized reaction conditions, 60 mM 3-cyanopyridine was converted to nicotinic acid in 15 min, which indicated ZJB-09150 has potentials of application in large scale production of nicotinic acid.  相似文献   

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