Advances in bacterial adsorption and transport of aromatic compounds / 生物工程学报

Aromatic compounds are a class of organic compounds with benzene ring(s). Aromatic compounds are hardly decomposed due to its stable structure and can be accumulated in the food cycle, posing a great threat to the ecological environment and human health. Bacteria have a strong catabolic ability to degrade various refractory organic contaminants (e.g., polycyclic aromatic hydrocarbons, PAHs). The adsorption and transportation are prerequisites for the catabolism of aromatic compounds by bacteria. While remarkable progress has been made in understanding the metabolism of aromatic compounds in bacterial degraders, the systems responsible for the uptake and transport of aromatic compounds are poorly understood. Here we summarize the effect of cell-surface hydrophobicity, biofilm formation, and bacterial chemotaxis on the bacterial adsorption of aromatic compounds. Besides, the effects of outer membrane transport systems (such as FadL family, TonB-dependent receptors, and OmpW family), and inner membrane transport systems (such as major facilitator superfamily (MFS) transporter and ATP-binding cassette (ABC) transporter) involved in the membrane transport of these compounds are summarized. Moreover, the mechanism of transmembrane transport is also discussed. This review may serve as a reference for the prevention and remediation of aromatic pollutants.

Polycyclic aromatic hydrocarbons degradation by marine-derived basidiomycetes: optimization of the degradation process

ABSTRACT Pyrene and benzo[a]pyrene (BaP) are high molecular weight polycyclic aromatic hydrocarbons (PAHs) recalcitrant to microbial attack. Although studies related to the microbial degradation of PAHs have been carried out in the last decades, little is known about degradation of these environmental pollutants by fungi from marine origin. Therefore, this study aimed to select one PAHs degrader among three marine-derived basidiomycete fungi and to study its pyrene detoxification/degradation. Marasmiellus sp. CBMAI 1062 showed higher levels of pyrene and BaP degradation and was subjected to studies related to pyrene degradation optimization using experimental design, acute toxicity, organic carbon removal (TOC), and metabolite evaluation. The experimental design resulted in an efficient pyrene degradation, reducing the experiment time while the PAH concentration applied in the assays was increased. The selected fungus was able to degrade almost 100% of pyrene (0.08 mg mL-1) after 48 h of incubation under saline condition, without generating toxic compounds and with a TOC reduction of 17%. Intermediate metabolites of pyrene degradation were identified, suggesting that the fungus degraded the compound via the cytochrome P450 system and epoxide hydrolases. These results highlight the relevance of marine-derived fungi in the field of PAH bioremediation, adding value to the blue biotechnology.

Draft genome of Nocardia farcinica TRH1, a linear and polycyclic aromatic hydrocarbon-degrading bacterium isolated from the coast of Trindade Island, Brazil

Abstract Here, we report the draft genome sequence and annotation of Nocardia farcinica TRH1, a petroleum hydrocarbons degrading Actinobacteria isolated from the coastal water of Trindade Island, Brazil.

Degradation of polynuclear aromatic hydrocarbons by two strains of Pseudomonas

ABSTRACT The goal of this investigation was to isolate competent polynuclear aromatic hydrocarbons degraders that can utilize polynuclear aromatic hydrocarbons of former industrial sites at McDoel Switchyard in Bloomington, Indiana. Using conventional enrichment method based on soil slurry, we isolated, screened and purified two bacterial species strains PB1 and PB2. Applying the ribotyping technique using the 16S rRNA gene analysis, the strains were assigned to the genus Pseudomonas (Pseudomonas plecoglossicida strain PB1 and Pseudomonas sp. PB2). Both isolates showed promising metabolic capacity on pyrene sprayed MS agar plates during the preliminary investigations. Using time course studies in the liquid cultures at calculated concentrations 123, 64, 97 and 94 ppm for naphthalene, chrysene, fluroanthene and pyrene, P. plecoglossicida strain PB1 and Pseudomonas sp. PB2 showed partial utilization of the polynuclear aromatic hydrocarbons. Naphthalene was degraded between 26% and 40%, chrysene 14% and 16%, fluroanthene 5% and 7%; pyrene 8% and 13% by P. plecoglossicida strain PB1 and Pseudomonas sp. PB2 respectively. Based on their growth profile, we developed a model R2 = 1 to predict the degradation rate of slow polynuclear aromatic hydrocarbon-degraders where all the necessary parameters are constant. From this investigation, we confirm that the former industrial site soil microbial communities may be explored for the biorestoration of the industrial site.

Sphingomonas from petroleum-contaminated soils in Shenfu, China and their PAHs degradation abilities

Abstract Members of the Sphingomonas genus are often isolated from petroleum-contaminated soils due to their unique abilities to degrade polycyclic aromatic hydrocarbons (PAHs), which are important for in situ bioremediation. In this study, a combined phenotypic and genotypic approach using streptomycin-containing medium and Sphingomonas -specific PCR was developed to isolate and identify culturable Sphingomonas strains present in petroleum-contaminated soils in the Shenfu wastewater irrigation zone. Of the 15 soil samples examined, 12 soils yielded yellow streptomycin-resistant colonies. The largest number of yellow colony-forming units (CFUs) could reach 105 CFUs g-1 soil. The number of yellow CFUs had a significant positive correlation (p < 0.05) with the ratio of PAHs to total petroleum hydrocarbons (TPH), indicating that Sphingomonas may play a key role in degrading the PAH fraction of the petroleum contaminants at this site. Sixty yellow colonies were selected randomly and analyzed by colony PCR using Sphingomonas -specific primers, out of which 48 isolates had PCR-positive signals. The 48 positive amplicons generated 8 distinct restriction fragment length polymorphism (RFLP) patterns, and 7 out of 8 phylotypes were identified as Sphingomonas by 16S rRNA gene sequencing of the representative strains. Within these 7 Sphingomonas strains, 6 strains were capable of using fluorene as the sole carbon source, while 2 strains were phenanthrene-degrading Sphingomonas. To the best of our knowledge, this is the first report to evaluate the relationship between PAHs contamination levels and culturable Sphingomonas in environmental samples.

Genotoxicidade in vitro das frações orgânica e solúvel em água de material particulado de ar em três locais do Estado de São Paulo / Genotoxicity in vitro of organic and soluble water fractions of airborne particulate matter in three sites of São Paulo State

O aumento na poluição ambiental é na atualidade uma das grandes preocupações em nível mundial. Especificamente, a poluição atmosférica por material particulado tem demostrado ser um fator determinante no desenvolvimento de doenças cardiopulmonares e câncer de pulmão nas populações expostas. O material particulado é constituído por uma mistura complexa de compostos orgânicos e inorgânicos, muitos dos quais possuem potencial mutagênico e genotóxico. As características destes compostos variam em função da suas propriedades físicas, químicas e em razão das condições meteorológicas prevalecentes. A maior parte dos estudos tem se focado em avaliar o potencial genotóxico da fração orgânica do material particulado e poucos estudos têm explorado a fração solúvel em água e a contribuição diferencial das diversas espécies químicas presentes nesta fração para o dano genotóxico. O objetivo deste trabalho foi avaliar os efeitos mutagênicos e genotóxicos in vitro da fração orgânica e da fração solúvel em água de material particulado (MP10) coletado em três locais diferentes do estado de São Paulo e estabelecer a relação entre a composição química e o efeito biológico observado. Para isto, realizou­-se a extração orgânica e solúvel em água de 12 amostras de MP10. A mutagenicidade e genotoxicidade foram avaliadas usando o ensaio de Salmonella/microssoma e o teste de micronúcleos (MN) em células A549 e 16 hidrocarbonetos Policíclicos Aromáticos (HPAs) e 15 metais hidrossolúveis presentes nas amostras foram determinados quimicamente. Adicionalmente, foi determinada a metodologia de extração da fracção solúvel em água e se avaliou a estabilidade química e biológica desta fração. Os resultados indicam que a extração assistida por micro-ondas é um método eficiente para a extração da fração solúvel em agua do MP e que um tempo superior a 60 dias de armazenamento e congelamento deste tipo de extrato tem um efeito significativo sobre os resultados analíticos e a resposta biológica. Foi demonstrado ainda que as duas frações de MP estudadas são responsáveis pela indução do dano ao DNA e que não existe uma relação direta entre a concentração de MP e o efeito genotóxico observado, confirmando a importância do uso de bioensaios na avaliação da genotoxicidade de misturas complexas como o MP. Os HPAs prevalecentes nas amostras de PM10 foram fluorantene e benzo(ghi)perileno. Nos extratos solúveis em água, as maiores concentrações de metais foram determinadas para zinco, ferro e cobre. Confirmou-se que a indução de MN e o ensaio de Salmonella/microssoma representam uma poderosa ferramenta na avaliação da polução atmosférica e que as análises químicas por si só não são suficientes para a proteção e predição dos efeitos biológicos em populações expostas

The increase of environmental pollution is today one a major concern worldwide. Specifically, air pollution by particulate matter has been shown to be a determining factor in the development of cardiopulmonary disease and lung cancer in exposed populations. The particulate material consists of a complex mixture of organic and inorganic compounds, many of which have mutagenic and genotoxic activity. The characteristics of these compounds vary according to their physical and chemical properties and also to the prevailing weather conditions. Most studies have focused on evaluating the genotoxic potential of the organic fraction of particulate material, but few studies have explored the water­-soluble fraction, and the differential contributions of different chemical species present in this fraction to genotoxic damage. The aim of this study was to evaluate the in vitro mutagenic and genotoxic effects of organic and water-soluble fractions of 12 samples of particulate matter (PM10) collected at three different sites in the state of São Paulo and establish the relationship between the chemical composition and the biological effect observed. The mutagenicity and genotoxicity were evaluated using the Salmonella/microssome test and the micronucleus assay (MN) in A549 cells and 16 Polycyclic Aromatic Hydrocarbons (PAHs) and 15 water-soluble metals present in the samples were chemically determined. Additionally, the extraction method of water- soluble fraction was determined and the chemical and biological stability of this fraction evaluated. The results indicate that the microwave­-assisted extraction is an efficient method for the extraction of the water-soluble compounds of PM and that the freezing and storage of the extract over 60 days has a significant effect on the mutagenic and analytical results of PM samples. It was demonstrated that the two PM fractions studied are responsible for the induction of DNA damage and that there is no direct relationship between the MP concentration and the genotoxic effect observed, confirming the importance of using bioassays in the genotoxicity evaluation of complex mixtures as PM. The PAHs prevailing in our samples were fluoranthene and benzo(ghi)perylene. In the water-soluble extracts, highest concentrations of the elements studied were found for zinc, iron, and copper in the three places of sampling. We confirmed that MN induction and Salmonella/microsome assay represents a powerful tool to evaluate the atmospheric air pollution and that the total concentration of PM and the chemical analyses alone would not be sufficient for the prognosis of biological effects in exposed populations

Morphological changes and growth of filamentous fungi in the presence of high concentrations of PAHs

In this study, we evaluated the effect of low and high molecular weight polycyclic aromatic hydrocarbons (PAHs), i.e., Phenanthrene, Pyrene and Benzo[a]pyrene, on the radial growth and morphology of the PAH-degrading fungal strains Aspergillus nomius H7 and Trichoderma asperellum H15. The presence of PAHs in solid medium produced significant detrimental effects on the radial growth of A. nomius H7 at 4,000 and 6,000 mg L⁻¹ and changes in mycelium pigmentation, abundance and sporulation ability at 1,000–6,000 mg L⁻¹. In contrast, the radial growth of T. asperellum H15 was not affected at any of the doses tested, although sporulation was observed only up to 4,000 mg L⁻¹ and as with the H7 strain, some visible changes in sporulation patterns and mycelium pigmentation were observed. Our results suggest that fungal strains exposed to high doses of PAHs significantly vary in their growth rates and sporulation characteristics in response to the physiological and defense mechanisms that affect both pigment production and conidiation processes. This finding is relevant for obtaining a better understanding of fungal adaptation in PAH-polluted environments and for developing and implementing adequate strategies for the remediation of contaminated soils.

Bioremediation of polyaromatic hydrocarbons (PAHs) using rhizosphere technology

The remediation of polluted sites has become a priority for society because of increase in quality of life standards and the awareness of environmental issues. Over the past few decades there has been avid interest in developing in situ strategies for remediation of environmental contaminants, because of the high economic cost of physicochemical strategies, the biological tools for remediation of these persistent pollutants is the better option. Major foci have been considered on persistent organic chemicals i.e. polyaromatic hydrocarbons (PAHs) due to their ubiquitous occurrence, recalcitrance, bioaccumulation potential and carcinogenic activity. Rhizoremediation, a specific type of phytoremediation that involves both plants and their associated rhizospheric microbes is the creative biotechnological approach that has been explored in this review. Moreover, in this review we showed the significance of rhizoremediation of PAHs from other bioremediation strategies i.e. natural attenuation, bioaugmentation and phytoremediation and also analyze certain environmental factor that may influence the rhizoremediation technique. Numerous bacterial species were reported to degrade variety of PAHs and most of them are isolated from contaminated soil, however few reports are available from non contaminated soil. Pseudomonas aeruginosa, Pseudomons fluoresens, Mycobacterium spp., Haemophilus spp., Rhodococcus spp., Paenibacillus spp. are some of the commonly studied PAH-degrading bacteria. Finally, exploring the molecular communication between plants and microbes, and exploiting this communication to achieve better results in the elimination of contaminants, is a fascinating area of research for future perspective.

A study of the status of exposure to polycyclic aromatic hydrocarbons (PAHs) in relation to its metabolites among workers in a Korean chemical factory / Um estudo sobre o estado de exposição a hidrocarbonetos aromáticos policíclicos (HAPs), em relação aos seus metabolitos, entre os trabalhadores em fábricas de produtos químicos coreana

The study was conducted to evaluate the status of worker exposure to polycyclic aromatic hydrocarbons (PAHs) through the measurement of urinary metabolites such as 1-hydroxypyrene (OHP) and 2-naphthol. A survey using a questionnaire involving 326 workers with measurement of urinary metabolites of 1-OHP and 2-naphthol was conducted. The differences in urinary 1-OHP and 2-naphthol concentrations, and changes in work, smoking habits and lifestyle were analyzed. The number of male subjects was 314 (96.3%), the largest age group was the fifth decade (170 cases, 52.1%). The urinary 1-OHP and 2-naphthol concentrations were significantly higher in the production workers. The urinary 1-OHP and 2-naphthol concentrations were significantly higher in smokers. In a multiple regression model, log (1-OHP) increased in smokers and production workers, while log (2-naphthol) only increased in smokers. Our results suggest that workers in this factory were exposed to PAHs from non-occupational as well as occupational sources. The occupational exposure to PAHs can be reduced through the improvement of the process, but the exposure due to smoking can be prevented only by giving up smoking.

O presente estudo foi realizado para avaliar o estado de exposição a hidrocarbonetos aromáticos policíclicos (HAPs) em trabalhadores, por medição de metabólitos urinários, tais como 1-hidroxipireno (OHP) e 2-naftol. Foi realizada uma pesquisa por questionário envolvendo 326 trabalhadores e a mensuração dos metabólitos urinários de 1-OHP e 2-naftol. Foram analisadas as diferenças na urinária 1-OHP e as concentrações de 2-naftol e mudanças pelo trabalho, hábito de fumar e estilo de vida. O número de indivíduos do sexo masculino foi de 314 (96,3%), a maior faixa etária foi a quinta década (170 casos, 52,1 %). Com relação aos metabólitos urinários 1 -OHP e 2-naftol, as concentrações foram significativamente maiores nos trabalhadores produtivos. As concentrações dos metabólitos urinários 1-OHP e 2-naftol foram significativamente maiores nos fumantes. Em um modelo de regressão múltipla, log (1-OHP) aumentou em fumantes e em trabalhadores produtivos, enquanto que log (2-naftol) aumentou apenas em fumantes. Nossos resultados sugerem que os trabalhadores desta fábrica foram expostos tanto a HAPs de fontes não ocupacionais como ocupacionais. A exposição ocupacional a HAPs pode ser diminuída através da melhoria do processo, mas a exposição devido ao fumo só pode ser impedida interrompendo esse hábito.

Detection of polycyclic aromatic hydrocarbon exposure from automobile exhaust fumes using urinary 1-hydroxypyrene level as an index.

Polycyclic aromatic hydrocarbons (PAHs) are hydrocarbon compounds which originate from incomplete combustion. In humans, PAHs are bioactivated to reactive metabolites which can bind covalently to DNA and subsequently initiate mutation and carcinogenesis. The measurement of PAHs exposure may be used as an index to classify a cancer risk group. The purpose of the present study was to measure the level of urinary 1-hydroxypyrene, a metabolite of PAHs, in subjects exposed to automobile exhaust fumes compared to non-exposed subjects. A urine sample was collected from each individual subject after the end of a working day and quantitated for 1-hydroxypyrene and creatinine by HPLC and spectrophotometric method, respectively. The results showed that average urinary 1-hydroxypyrene level in exposed subjects was significantly higher than non-exposed subjects (mean +/- SD of 0.0035 +/- 0.0032 and 0.0011 +/- 0.0010 micromol/l, respectively; P = 0.000). Average urinary creatinine level in exposed subjects was also significantly higher than non-exposed subjects (mean +/- SD of 0.01 +/- 0.005 and 0.008 +/- 0.006 mol/l, respectively; P = 0.040). The ratio of urinary 1-hydroxypyrene/mol creatinine level, of the exposed subjects was significantly higher than that of the non-exposed subjects (mean +/- SD of 0.37 +/- 0.28 and 0.19 +/- 0.22 micromol/mol creatinine, respectively; P = 0.002). CONCLUSION: Automobile exhaust fume exposed subjects have a higher risk to be exposed to PAHs than the non-exposed subjects. Urinary 1-hydroxypyrene level can be used as an index for an exposure of PAHs which have originated from automobile exhaust fumes and other sources as well.

Metabolism of polycyclic aromatic hydrocarbons by Aspergillus niger.

Aspergillus niger metabolised polycyclic aromatic hydrocarbons such as naphthalene, anthracene and phenanthrene. The maximum weight of mycelium was obtained at 8-10 days of incubation. TLC and HPLC analysis revealed the accumulation of metabolites in resting cell culture. The metabolism of naphthalene and anthracene follows the gentisate pathway, where as phenenthrene was metabolised via protocatechuate in this organism.