Degradation of gasoline aromatic hydrocarbons by two N2-fixing soil bacteria

Two bacterial strains, 3A and 5A, isolated from soil, were selected for their ability to degrade gasoline aromatic compounds and to fix N₂. Strains 3A and 5A have been ascribed to the genera Agrobacterium and Alcaligenes, respectively. Using gasoline as the sole carbon source these strains were as effective at degrading benzene, toluene and xylene as Pseudomonas putida ATCC12236, a reference biodegrading strain.     

微生物降解多环芳烃的研究进展

多环芳烃是一类长久存在于环境中,具有毒性、致突变与致癌等特性的环境优先污染物。本文对降解多环芳烃的微生物类群进行了阐述,介绍了在土壤与厌氧条件下细菌降解多环芳烃的研究情况,最后介绍了降解多环芳烃的相关酶类以及分子生物学的研究,并对消除环境中多环芳烃的相关生物技术提出展望。     

污染土壤中多环芳烃生物降解的调控研究

选用温度、湿度、表面活性剂ＴＷ８０和ＣＮＰ比４个因素为调控因子,采用正交法进行周期为１５０天的实验研究．结果表明,３０天后,土壤中ＰＡＨｓ的降解率可达４４．５～７４．６％,６０天后,达７０．４～９３．７％,降解率的不同与调控条件显著相关．在此期间,降解最佳条件为４０℃,湿度２５％,ＣＮＰ比为１２０１０１,ＴＷ８０分别为２００～５００ｍｇ·ｋｇ－１．实验结束时,土壤中ＰＡＨｓ的降解率达９１．２～９９．８％．降解的最佳条件是４０℃,湿度１５％．经Ｒ值判别表明,不同时期各因子对ＰＡＨｓ降解影响有所不同．温度对ＰＡＨｓ降解影响较大,表面活性剂对土壤中ＰＡＨｓ的生物降解有调控作用．     

Revealing potential functions of VBNC bacteria in polycyclic aromatic hydrocarbons biodegradation

The bioremediation of polycyclic aromatic hydrocarbon (PAH)‐contaminated sites is not running smoothly, because of the lower activity of PAH‐degrading bacteria in actual bioremediation applications. The phenomenon of “viable but nonculturable” (VBNC) state may be a main limiting factor for their poor biodegradation capabilities of PAHs. Due to their abilities of entering into the VBNC state, most of bacterial populations with PAH‐degradation potential remain unculturable. Resuscitation of VBNC bacteria will enhance the degradation capability of indigenous bacteria which will eventually obtain their better capabilities in environmental bioremediation. Although evidences have been presented indicating that resuscitation of VBNC bacteria in polychlorinated biphenyl (PCB)‐contaminated environments not only significantly enhanced PCB degradation, but also obtained novel highly efficient PCB‐degrading bacteria, scanty information is available on the VBNC bacteria in PAH‐contaminated sites. VBNC bacteria, as a vast majority of potential microbial resource could be the repository of novel highly efficient PAH‐biodegraders. Therefore, studies need to be done on resuscitation of VBNC bacteria to overcome key bottlenecks in bioremediation of PAH‐contaminated sites. This mini‐review provides a new insight into the potential functions of VBNC bacteria in PAHs biodegradation.

Significance and Impact of the Study

As the vast majority microbial resource, viable but nonculturable (VBNC) bacteria, which showed their potential functions in polycyclic aromatic hydrocarbons (PAHs) biodegradation, can be of great significance in environmental bioremediation. It is therefore important to resuscitate VBNC bacteria for their better capabilities. Meanwhile, preventing the indigenous functional community from entering into the VBNC state will also maintain the high activity of PAH‐degrading bacteria in actual bioremediation applications. Undoubtedly, much more work needs to be done to reveal indigenous micro‐organisms in the VBNC state from the perspective of environmental functions.     

Induction of aldehyde dehydrogenase by polycyclic aromatic hydrocarbons in rats

Short-term intragastric administration of selected polycyclic aromatic hydrocarbons (100 mg/kg daily for 4 days) to male Wistar rats resulted in marked changes in liver cytosolic aldehyde dehydrogenase activity. Non-carcinogenic anthracene, phenanthrene and chrysene produced a 2.5–3-fold increase in the activity assayed with propionaldehyde as substrate and NAD as coenzyme. Weakly carcinogenic 1,2-benzanthracene enhanced aldehyde dehydrogenase activity 9-fold and the potent carcinogens 3,4-benzpyrene and 3-methylcholanthrene 30-fold. With benzaldehyde as substrate and NADP as coenzyme the differences between the groups were even more pronounced. Somewhat similar but less manifest effects on aldehyde dehydrogenase activity were detected also in the liver microsomes and in the postmitochondrial fractions of the small intestinal mucosa. On the basis of their ability to induce aldehyde dehydrogenase activity the compounds could be divided into three groups. This classification was found to correlate well with the carcinogenic potency of the compounds. It appeared that the exposure to polycyclic aromatic hydrocarbons, especially the carcinogenic ones, was followed by synthesis of a new aldehyde dehydrogenase form. This new form was differentiated from the normally existing cytosolic aldehyde dehydrogenase by its ability to oxidize benzaldehyde in the presence of NADP.     

Utilization of aromatic compounds by phototrophic purple nonsulfur bacteria

Biodegradation of aromatic compounds byRhodopseudomonas blastica andRhodospirillum rubrum appears to be lacking in the literature. The above species grew phototrophically (illuminated anaerobic conditions) on a variety of organic compounds. They were found to degrade benzoate, benzyl alcohol, 4-hydroxy-3,5-dimethoxybenzoate (Syringate) and 4-hydroxy-3-methoxybenzoate (vanillate). The ability of the above species to photocatabolize aromatic compounds indicates that these organisms may be ecologically significant as scavengers of aromatic derivatives in illuminated anaerobic habitats in nature.     

Utilization of monocyclic aromatic hydrocarbons individually and in mixture by bacteria isolated from petroleum-contaminated soil

The fate of benzene, ethylbenzene, toluene, xylenes (BTEX) compounds through biodegradation was investigated using two different bacteria, Ralstonia picketti (BP-20) and Alcaligenes piechaudii (CZOR L-1B). These bacteria were isolated from extremely polluted soils contaminated with petroleum hydrocarbons. PCR and Fatty Acid Methyl Ester (FAME) were used to identify the isolates. In this study, BTEX biodegradation, applied as a mixture or as individual compounds by the bacteria was evaluated. Both bacteria were shown to degrade each of the BTEX compounds individually and in mixture. However, Alcaligenes piechaudii was a better degrader of BTEXs both in the mixture and individually. Differences between BTEX biodegradation in the mixture and individually were observed, especially in the case of benzene. The degradation of all BTEXs in the mixture was lower than the degradation of individual compounds for both bacteria tested. In the all experiments, toluene and m + p- xylenes were better removed than the other BTEXs. No intermediates of biodegradation were detected. Biosurfactant production was observed by culture techniques. In addition, 3-hydroxy fatty acids, important in biosurfactant production, were observed by FAME analysis. The test results indicate that the bacteria could contribute to bioremediation of aromatic hydrocarbon pollution.     

A novel bioreactor for the biodegradation of inhibitory aromatic solvents: Experimental results and mathematical analysis

A novel bioreactor for the biodegradation of toxic aromatic solvents, such as benzene, toluene, and xylenes in liquid effluent stream, was developed. Silicon tubing was immersed in the completely mixed and aerated bioreactor, and liquid toluene as a model solvent was circulated within the tubing. Toluene diffused out of the tube wall and was transferred at high rate into the culture broth, where biodegradation occurred. The effect of operating parameters on the toluene transfer rate was investigated. During continuous operation, the biodegradation rate was considerably higher than those obtained using conventional methods. A mathematical model was established for continuous biodegradation, and simulation results coincided with the experimental results. The performance and operational criteria of the bioreactor were analyzed on the basis of both the experimental and simulation results. (c) 1992 John Wiley & Sons, Inc.     

Metabolism of alkylbenzenes, alkanes, and other hydrocarbons in anaerobic bacteria

Aromatic and aliphatic hydrocarbons are the main constituents of petroleum and its refined products. Whereas degradation of hydrocarbons by oxygen-respiring microorganisms has been known for about a century, utilization of hydrocarbons under anoxic conditions has been investigated only during the past decade. Diverse strains of anaerobic bacteria have been isolated that degrade toluene anaerobically, using nitrate, iron(III), or sulfate as electron acceptors. Also, other alkylbenzenes such as m-xylene or ethylbenzene are utilized by a number of strains. The capacity for anaerobic utilization of alkylbenzenes has been observed in members of the -, -, - and -subclasses of the Proteobacteria. Furthermore, denitrifying bacteria and sulfate-reducing bacteria with the capacity for anaerobic alkane degradation have been isolated, which are members of the - and -subclass, respectively. The mechanism of the activation of hydrocarbons as apolar molecules in the absence of oxygen is of particular interest.The biochemistry of anaerobic toluene degradation has been studied in detail. Toluene is activated by addition to fumarate to yield benzylsuccinate, which is then further metabolized via benzoyl-CoA. The toluene-activating enzyme presents a novel type of glycine radical protein. Another principle of anaerobic alkylbenzene activation has been observed in the anaerobic degradation of ethylbenzene. Ethylbenzene in denitrifying bacteria is dehydrogenated to 1-phenylethanol and further to acetophenone; the latter is also metabolized to benzoyl-CoA. Naphthalene is presumably activated under anoxic conditions by a carboxylation reaction. Investigations into the pathway of anaerobic alkane degradation are only at the beginning. The saturated hydrocarbons are mostlikely activated by addition of a carbon compound rather than by desaturation and hydration, as speculated about in some early studies. An anaerobic oxidation of methane with sulfate as electron acceptor has been documented in aquatic sediments. The process is assumed to involve a reversal of methanogenesis catalyzed by Archaea, and scavenge of an electron-carrying metabolite by sulfate-reducing bacteria. Among unsaturated non-aromatic hydrocarbons, anaerobic bacterial degradation has been demonstrated and investigated with n-alkenes, alkenoic terpenes and the alkyne, acetylene.     

Degradation of volatile hydrocarbons from steam-classified solid waste by a mixture of aromatic hydrocarbon-degrading bacteria

Steam classification is a process for treatment of solid waste that allows recovery of volatile organic compounds from the waste via steam condensate and off-gases. A mixed culture of aromatic hydrocarbon-degrading bacteria was used to degrade the contaminants in the condensate, which contained approx. 60 hydrocarbons, of which 38 were degraded within 4 d. Many of the hydrocarbons, including styrene, 1,2,4-trimethylbenzene, naphthalene, ethylbenzene, m-/p-xylene, chloroform, 1,3-dichloropropene, were completely or nearly completely degraded within one day, while trichloroethylene and 1,2,3-trichloropropane were degraded more slowly.     

A rapid screening method for bacteria degrading polycyclic aromatic hydrocarbons

Aims:  A rapid procedure was developed to screen for bacteria that are able to grow on polycyclic aromatic hydrocarbons (PAHs).
Methods and Results:  A drop of ethyl ether-dissolved PAH is spread on a sterilized cellulose acetate/nitrate filter lying on the top of a mineral salts agarose plate. After the evaporation of ethyl ether, a serially diluted sample is spread over the filter and incubated. Subsequently, the PAH degrading bacteria can be counted and isolated.
Conclusions:  This procedure is a simple method for screening bacterial isolates for the ability to grow with PAHs.
Significance and Impact of the Study:  This technique is rapid to screen and/or count PAH-degrading bacteria and is also used to streak cultivation without disrupting the PAH layer on plate.     

Biodegradation of polycyclic aromatic hydrocarbons by Pichia anomala

Pichia anomala 2.2540, isolated from soil contaminated by crude oil, degraded naphthalene, dibenzothiophene, phenanthrene and chrysene, both singly and in combination. The yeast degraded 4.5 mg naphthalene l(-1) within 24 h. Phenanthrene was degraded after a lag of 24 h. When a mixture of all four polycyclic aromatic hydrocarbons was treated at either 0.1-1.6 mg l(-1) or 3.1-5.3 mg l(-1), naphthalene was completely degraded first within 24 h, followed by phenanthrene and dibenzothiophene after 48 h. Chrysene, which remained in the mixture even after 96 h, could be degraded along with naphthalene. Chrysene at 0.7 and 1 mg l(-1), in the presence of 4.3 and 65 mg naphthalene l(-1), respectively, was removed within 96 h.     

Sulphate-reducing bacteria,palladium and the reductive dehalogenation of chlorinated aromatic compounds

The surfaces of cells of Desulfovibrio desulfuricans,Desulfovibrio vulgaris and a new strain, Desulfovibrio sp. `Oz-7' were used to manufacturea novel bioinorganic catalyst via the reduction of Pd(II) to Pd(0) at the cell surface usinghydrogen as the electron donor. The ability of the palladium coated (palladised) cells to reductivelydehalogenate chlorophenol and polychlorinated biphenyl species was demonstrated. Dried, palladisedcells of D. desulfuricans, D. vulgaris and Desulfovibrio sp. `Oz-7'were more effective bioinorganic catalysts than Pd(II) reduced chemically under H₂ orcommercially available finely divided Pd(0). Differences were observed in the catalyticactivity of the preparations when compared with each other. Negligible chloride release occurredfrom chlorophenol and polychlorinated biphenyls using biomass alone.     

Colorimetric assays for biodegradation of polycyclic aromatic hydrocarbons by fungal laccases

Polycyclic aromatic hydrocarbons (PAHs) are highly toxic organic pollutants widely distributed in terrestrial and aquatic environments. In the present work, 2 colorimetric assays for laccase-catalyzed degradation of PAHs were developed based on studies of the oxidation of 12 aromatic hydrocarbons by fungal laccases from Trametes versicolor and Myceliophthora thermophila. Using a sodium borohydride water-soluble solution, the authors could reduce the single product of laccase-catalyzed anthracene biooxidation into the orange-colored 9,10-anthrahydroquinone, which is quantifiable spectrophotometrically. An assay using polymeric dye (Poly R-478) as a surrogate substrate for lignin degradation by laccase in the presence of mediator is also presented. The decolorization of Poly R-478 was correlated to the oxidation of PAHs mediated by laccases. This demonstrates that a ligninolytic indicator such as Poly R-478 can be used to screen for PAH-degrading laccases; it will also be useful in screening mutant libraries in directed evolution experiments. Poly R-478 is stable and readily soluble. It has a high extinction coefficient and low toxicity toward white rot fungi, yeast, and bacteria, which allow its application in a solid-phase assay format.     

Aerobic biodegradation of methyl tert-butyl ether (MTBE) by pure bacterial cultures isolated from contaminated soil

Summary Methyl tert-Butyl Ether (MTBE) has been used in gasoline as a substitute for lead-based additives, which have been demonstrated to be toxic. MTBE however, is persistent in soil and water, showing high affinity for water and low affinity for soil, and has become an important contaminant. Therefore, the aim of this work was to isolate and identify soil microorganisms capable of degrading MTBE. Two samples were taken from a gasoline-contaminated soil at a service station and 59 different bacterial strains were isolated by enrichment culture with three consecutive selective transfers. Biochemical and morphological characterization of the bacterial isolates classified them into the following groups: Bacillus, Rhodococcus, Micrococcus, Aureobacterium and Proteus. Twelve strains were selected for evaluation of MTBE biodegradation depending on visual growth and biomass production of the isolates in minimal salt broth. Six strains significantly reduced MTBE concentration (22–37%) compared to an abiotic control after 5 days of incubation. Although it has been considered that MTBE is degraded mainly by cometabolism, our results demonstrate that these microorganisms are able to reduce MTBE concentration when MTBE is the sole source of carbon.     

Biodegradation of central intermediate compounds produced from biodegradation of aromatic compounds

In this study I consider the incomplete biodegradation of aromatic compounds during the wastewater cycle between aerobic or anaerobic zones in biological nutrient removal processes, including aerobic biodegradation of compounds (such as cyclohex-1-ene-1-carboxyl-CoA) produced during the incomplete anaerobic biodegradation of aromatic compounds, and anaerobic biodegradation of compounds (such as catechol, protocatechuate, and gentisic acid) produced during the incomplete aerobic biodegradation of aromatic compounds. Anaerobic degradation of the aerobic central intermediates that result from the incomplete aerobic degradation of aromatic compounds usually leads to benzoyl-CoA. On the other hand, aerobic degradation of the anaerobic central intermediates that result from the incomplete anaerobic degradation of aromatic compounds usually leads to protocatechuate.