Biodegradation Of Di-(2-ethylhexyl) Phthalate Research Articles

A novel and efficient strategy for the biodegradation of di(2-ethylhexyl) phthalate by Fusarium culmorum.

Di(2-ethylhexyl) phthalate (DEHP) is a plasticizer that is used worldwide and raises concerns because of its prevalence in the environment and potential toxicity. Herein, the capability of Fusarium culmorum to degrade a high concentration (3 g/L) of DEHP as the sole carbon and energy source in solid-state fermentation (SSF) was studied. Cultures grown on glucose were used as controls. The biodegradation of DEHP by F. culmorum reached 96.9% within 312 h. This fungus produced a 3-fold higher esterase activity in DEHP-supplemented cultures than in control cultures (1288.9 and 443.2 U/L, respectively). In DEHP-supplemented cultures, nine bands with esterase activity (24.6, 31.2, 34.2, 39.5, 42.8, 62.1, 74.5, 134.5, and 214.5 kDa) were observed by zymography, which were different from those in control cultures and from those previously reported for cultures grown in submerged fermentation. This is the first study to report the DEHP biodegradation pathway by a microorganism grown in SSF. The study findings uncovered a novel biodegradation strategy by which high concentrations of DEHP could be biodegraded using two alternative pathways simultaneously. F. culmorum has an outstanding capability to efficiently degrade DEHP by inducing esterase production, representing an ecologically promising alternative for the development of environmental biotechnologies, which might help mitigate the negative impacts of environmental contamination by this phthalate. KEY POINTS: •F. culmorum has potential to tolerate and remove di(2-ethylhexyl) phthalate (DEHP) •Solid-state fermentation is an efficient system for DEHP degradation by F. culmorum •High concentrations of DEHP induce high levels of esterase production by F. culmorum.

New mechanisms of biochar-assisted vermicomposting by recognizing different active di-(2-ethylhexyl) phthalate (DEHP) degraders across pedosphere, charosphere and intestinal sphere

Biochar-assisted vermicomposting can significantly accelerate soil DEHP degradation, but little information is known about the underlying mechanisms as different microspheres exist in soil ecosystem. In this study, we identified the active DEHP degraders in biochar-assisted vermicomposting by DNA stable isotope probing (DNA-SIP) and surprisingly found their different compositions in pedosphere, charosphere and intestinal sphere. Thirteen bacterial lineages (Laceyella, Microvirga, Sphingomonas, Ensifer, Skermanella, Lysobacter, Archangium, Intrasporangiaceae, Pseudarthrobacter, Blastococcus, Streptomyces, Nocardioides and Gemmatimonadetes) were responsible for in situ DEHP degradation in pedosphere, whereas their abundance significantly changed in biochar or earthworm treatments. Instead, some other active DEHP degraders were identified in charosphere (Serratia marcescens and Micromonospora) and intestinal sphere (Clostridiaceae, Oceanobacillus, Acidobacteria, Serratia marcescens and Acinetobacter) with high abundance. In biochar-assisted vermicomposting, the majority of active DEHP degraders were found in charosphere, followed by intestinal sphere and pedosphere. Our findings for the first time unraveled the spatial distribution of active DEHP degraders in different microspheres in soil matrices, explained by DEHP dynamic adsorption on biochar and desorption in earthworm gut. Our work highlighted that charosphere and intestinal sphere exhibited more contribution to the accelerated DEHP biodegradation than pedosphere, providing novel insight into the mechanisms of biochar and earthworm in improving contaminant degradation.

Biodegradation of di(2-ethylhexyl) phthalate by a new bacterial consortium.

Di(2-ethylhexyl) phthalate (DEHP) with continuous high concentration was used as the sole carbon and energy source to isolate a new bacterial consortium (K1) from agricultural soil covered with plastic film for a long time. Unclassified Comamonadaceae, Achromobacter, and Pseudomonas in K1 were identified as major genera of the consortium by high-throughput sequencing, and unclassified Commanadaceae was first reported to be related to DEHP degradation. Response surface method (RSM) showed that the optimum conditions for K1 to degrade DEHP were 31.4 °C, pH 7.3, and a concentration of 420 mg L-1. K1 maintains normal cell viability and stable DEHP degradation efficiency in the range of 10-3000 mg L-1 DEHP concentration, which is superior to existing research. The biodegradation of DEHP followed first-order kinetics when the initial concentration of DEHP was between 100 and 3,000 mg L-1. GC-MS analysis of different treatment groups showed that DEHP was degraded by the consortium group through the de-esterification pathway, and treatment effect was significantly better than that of the single bacteria treatment group. The subsequent substrate utilization experiment further confirmed that K1 could quickly mineralize DEHP. In addition, K1 has high degradation capacity for the most common phthalate acid esters in the environment.

Fungal Bioremediation of the Plasticizer Hazardous Compound di-2-Ethylhexyl Phthalate (DEHP) in Urine and Blood Bags

The hazardous compound di-2-ethylhexyl phthalate (DEHP) is widely used as polyvinyl chloride plasticizer. The present research studied the fungal biodegradation of DEHP contained in blood and urine bags. Soil-plate method was used for fungal isolation from heavily plastic polluted soil using Martin’s and Sabouraud’s agar media, where DEHP was the sole carbon source. Isolated fungal species were identified morphologically according to Moubasher (Soil Fungi in Qatar and Arab Countries University of Qatar, Qatar The Center for Scientific and Research, 1993) as Aspergillus nidulans, Aspergillus niger and Rhizopus nigricans. DEHP concentrations were determined in 1 g of soil, urine bags and blood bags to be 0.92, 2.5 and 2.6 g/l, respectively. Samples of both bags (as a sole carbon source) were artificially inoculated with the isolated fungi and incubated for 20 days. As the time increased, the growth increased where Rhizopus nigricans obtained the highest dry weight in urine bags after 20 days of incubation, while A. nidulans had the highest dry weight in blood bags. Also, a sharp declining of initial pH (6.8) reached 4.7 in urine bags with A. niger growth, while reached 2.5 after A. nidulans growth in blood bags. DEHP% decreased as time increased indicating a continuous DEHP utilization by the three fungal species. Aspergillus niger was the most DEHP degrading fungal species in both bags. Scanning electron microscope examination showed an uniform plastic network in both bags before fungal treatment. While, a microporous network was observed on the plastic surfaces in both bags after fungal treatment due to DEHP utilization. The most DEHP metabolizing fungal species were further identified molecularly using internal transcribed spacer primers to be Aspergillus niger and Aspergillus nidulans with accession numbers MZ832174 and MT919276, respectively.

Adaptation of bacterial community in maize rhizosphere for enhancing dissipation of phthalic acid esters in agricultural soil.

Rhizospheric degradation is a green and in situ strategy to accelerate dissipation of organic pollutants in soils. However, the mechanism on microbial degradation of phthalic acid esters (PAEs) in rhizosphere is still unclear. Here, the bacterial community and function genes in bulk and rhizospheric soils of maize (Zea mays L.) exposed to gradient concentrations of di-(2-ethylhexyl) phthalate (DEHP) were analyzed with 16S rRNA, metagenomic sequencing and quantitative PCR (qPCR). Maize rhizosphere significantly increased the dissipation of DEHP by 4.02-11.5% in comparison with bulk soils. Bacterial community in rhizosphere exhibited more intensive response and shaped its beneficial structure and functions to DEHP stress than that in bulk soils. Both rhizospheric and pollution effects enriched more PAE-degrading bacteria (e.g., Bacillus and Rhizobium) and function genes in rhizosphere than in bulk soil, which played important roles in degradation of PAEs in rhizosphere. The PAE-degrading bacteria (including genera Sphingomonas, Sphingopyxis and Lysobacter) identified as keystone species participated in DEHP biodegradation. Identification of PAE intermediates and metagenomic reconstruction of PAE degradation pathways demonstrated that PAE-degrading bacteria degraded PAEs through cooperation with PAE-degrading and non-PAE-degrading bacteria. This study provides a comprehensive knowledge for the microbial mechanism on the superior dissipation of PAEs in rhizosphere.

Biodegradation of high di-(2-Ethylhexyl) phthalate (DEHP) concentration by food waste composting and its toxicity assessment using seed germination test

Di-(2-ethylhexyl) phthalate (DEHP), a plasticizer derived from phthalate ester, is used as an additive in industrial products such as plastics, paints, and medical devices. However, DEHP is known as an endocrine-disrupting chemical, causing cancers and adverse effects on human health. This study evaluated DEHP biodegradation efficiency via food waste composting during 35 days of incubation. At high DEHP concentrations (2167 mg kg−1) in food waste compost mixture, the DEHP biodegradation efficiency was 99% after 35 days. The highest degradation efficiency was recorded at the thermophilic phase (day 3 - day 11) with the biodegradation rate reached 187 mg kg−1 day−1. DEHP was metabolized to dibutyl phthalate (DBP) and dimethyl phthalate (DMP) and would be oxidized to benzyl alcohol (BA) and mineralized into CO2 and water via various metabolisms. Finally, the compost's quality with residual DEHP was evaluated using Brassica chinensis L. seeds via 96 h of germination tests. The compost (at day 35) with a trace amount of DEHP as the end product showed no significant effect on the germination rate of Brassica chinensis L. seeds (88%) compared to that without DEHP (94%), indicating that the compost can be reused as fertilizer in agricultural applications. These results provide an improved understanding of the DEHP biodegradation via food waste composting without bioaugmentation and hence facilitating its green remediation and conversion into value-added products. Nevertheless, further studies are needed on DEHP biodegradation in large-scale food waste composting or industrial applications.

Biochemical pathways and enhanced degradation of endocrine disruptor di-2-ethylhexyl phthalate by an indigenous isolate Bacillus sp. MY156

Di-2-ethylhexyl phthalate (DEHP) widely exists in the environment and has raised an increasing global concern. A bacterial strain capable of degrading DEHP was isolated from the activated sludge at a local wastewater treatment plant and identified as Bacillus sp. MY156. A nearly complete degradation of DEHP at 500 mg L−1 was reached within 5 days when MY156 was grown at pH 8 and 30 °C. The biodegradation of DEHP (50–600 mg L−1) followed the first-order kinetics. Mass balance analysis showed the efficient degradation of DEHP with the ratio of biomass to DEHP at 0.72. The dehydrogenase activity was positively related to the biodegradation of DEHP, while the extracellular polymeric substance (EPS) might have played a key role in overcoming the potential inhibition caused by DEHP. The microelements (Fe3+, Zn2+, and Mn2+) supplements stimulated the DEHP degradation while copper (Cu2+) showed inhibitory. The proposed intermediate metabolites of the DEHP degradation included mono-ethylhexyl phthalate, diethyl phthalate, dimethyl phthalate, phthalic acid, and protocatechuate. Together with the micro-morphological observation, biodegradation experiments in saline and different real water environments suggested the isolate possessing strong halo-tolerance and potential applicable ability to adapt to various environmental conditions. The isolate could be used as a potential and efficient phthalic acid esters (PAEs) degrader for the bioremediation of contaminated sites.

Biodegradation of di-(2-ethylhexyl) phthalate by novel Rhodococcus sp. PFS1 strain isolated from paddy field soil.

Di-(2-ethylhexyl)-phthalate (DEHP) is the phthalate ester frequently utilized as a plasticizer, commonly found in cosmetics, packaging materials; moreover, it has carcinogenic and mutagenic effects on humans. In the current study, we isolated the soil bacterium Rhodococcus sp. PFS1 and to assess its DEHP degradation ability in various environmental conditions. The strain PFS1 was isolated from paddy field soil and identified by the 16S rRNA sequencing analyses. The strain PFS1 was examined for its biodegradation ability of DEHP at various pH, temperature, salt concentration, glucose concentration, and high and low concentrations of DEHP. Moreover, the biodegradation of DEHP at a contaminated soil environment by strain PFS1 was assessed. Further, the metabolic pathway of DEHP degradation by PFS1 was analyzed by HPLC-MS analysis. The results showed that the strain PFS1 effectively degraded the DEHP at neutral pH and temperature 30°C; moreover, expressed excellent DEHP degradation at the high salt concentration (up to 50g/L). The strain PFS1 was efficiently degraded the different tested phthalate esters (PAEs) up to 90%, significantly removed the DEHP contamination in soil along with native organisms which are present in soil up to 94.66%; nevertheless, the PFS1 alone degraded the DEHP up to 87.665% in sterilized soil. According to HPLC-MS analysis, DEHP was degraded into phthalate (PA) by PFS1 strain via mono(2-ethylehxyl) phthalate (MEHP); then PA was utilized for cell growth. These results suggest that Rhodococcus sp. PFS1 has excellent potential to degrade DEHP at various environmental conditions especially in contaminated paddy field soil.

Integrated Multi-omics Investigations Reveal the Key Role of Synergistic Microbial Networks in Removing Plasticizer Di-(2-Ethylhexyl) Phthalate from Estuarine Sediments.

ABSTRACTDi-(2-ethylhexyl) phthalate (DEHP) is the most widely used plasticizer worldwide, with an annual global production of more than 8 million tons. Because of its improper disposal, endocrine-disrupting DEHP often accumulates in estuarine sediments in industrialized countries at submillimolar levels, resulting in adverse effects on both ecosystems and human beings. The microbial degraders and biodegradation pathways of DEHP in O2-limited estuarine sediments remain elusive. Here, we employed an integrated meta-omics approach to identify the DEHP degradation pathway and major degraders in this ecosystem. Estuarine sediments were treated with DEHP or its derived metabolites, o-phthalic acid and benzoic acid. The rate of DEHP degradation in denitrifying mesocosms was two times slower than that of o-phthalic acid, suggesting that side chain hydrolysis of DEHP is the rate-limiting step of anaerobic DEHP degradation. On the basis of microbial community structures, functional gene expression, and metabolite profile analysis, we proposed that DEHP biodegradation in estuarine sediments is mainly achieved through synergistic networks between denitrifying proteobacteria. Acidovorax and Sedimenticola are the major degraders of DEHP side chains; the resulting o-phthalic acid is mainly degraded by Aestuariibacter through the UbiD-dependent benzoyl coenzyme A (benzoyl-CoA) pathway. We isolated and characterized Acidovorax sp. strain 210-6 and its extracellular hydrolase, which hydrolyzes both alkyl side chains of DEHP. Interestingly, genes encoding DEHP/mono-(2-ethylhexyl) phthalate (MEHP) hydrolase and phthaloyl-CoA decarboxylase—key enzymes for side chain hydrolysis and o-phthalic acid degradation, respectively—are flanked by transposases in these proteobacterial genomes, indicating that DEHP degradation capacity is likely transferred horizontally in microbial communities.IMPORTANCE Xenobiotic phthalate esters (PAEs) have been produced on a considerably large scale for only 70 years. The occurrence of endocrine-disrupting di-(2-ethylhexyl) phthalate (DEHP) in environments has raised public concern, and estuarine sediments are major DEHP reservoirs. Our multi-omics analyses indicated that complete DEHP degradation in O2-limited estuarine sediments depends on synergistic microbial networks between diverse denitrifying proteobacteria and uncultured candidates. Our data also suggested that the side chain hydrolysis of DEHP, rather than o-phthalic acid activation, is the rate-limiting step in DEHP biodegradation within O2-limited estuarine sediments. Therefore, deciphering the bacterial ecophysiology and related biochemical mechanisms can help facilitate the practice of bioremediation in O2-limited environments. Furthermore, the DEHP hydrolase genes of active DEHP degraders can be used as molecular markers to monitor environmental DEHP degradation. Finally, future studies on the directed evolution of identified DEHP/mono-(2-ethylhexyl) phthalate (MEHP) hydrolase would bring a more catalytically efficient DEHP/MEHP hydrolase into practice.

Biodegradability of di-(2-ethylhexyl) phthalate by a newly isolated bacterium Achromobacter sp. RX

Di-(2-ethylhexyl) phthalate (DEHP) is a chemical plasticizer that has been commonly used in the manufacture of polyvinyl chloride. DEHP is one of the environmental pollution sources. In this study, a gram-negative strain RX bacterium utilizing DEHP as sole carbon source was isolated from activated sludge through screening test. This strain RX was identified as Achromobacter sp. RX based on its morphology, physiological properties and 16S rRNA gene sequence analysis. The results showed that the optimal conditions for the DEHP degradation were 30.0 °C and pH 7.0. The DEHP degradation induced by strain RX demonstrated nitrogen source dependent, while followed a decreasing degradation rate under the source of: NO3− > NH4+ > NO2−. The biodegradability of Achromobacter sp. RX was enhanced with Masson pine seed powder as a co-metabolic substrate and Tween-80 as a solubilizing agent. Meanwhile, the degrading kinetics analysis was performed in the condition of DEHP as sole carbon source. The DEHP degradation curves fitted well with the first-order kinetic model at 50–300 mg/L of DEHP, with the half-life ranging from 13.0 to 16.4 h. During the biodegradation of DEHP, mono-(2-ehtylhexyl) phthalate (MEHP) was firstly generated through de-esterification, followed by the formation of phthalic acid and benzoic acid after further de-esterification of MEHP. Benzoic acid was finally mineralized to CO2 and H2O. The decontamination of DEHP-contaminated soil by Achromobacter sp. RX was investigated using a rotating-drum bioreactor. Evolution of total organic carbon from the contaminated soil showed that 86.4%–91.7% of DEHP was mineralized at pH 7.0 and 30.0 °C within 96 h. Reusability of Achromobacter sp. RX and its lifetime were observed over six consecutive cycles. Thus, Achromobacter sp. RX possessed high DEHP biodegradability, which provided a good potential in dealing with DEHP-contaminated soil.

Enzyme mimics based membrane reactor for di(2-ethylhexyl) phthalate degradation

Di(2-ethylhexyl) phthalate (DEHP), the most abundantly used plasticizer, was considered to be a hazardous chemical that was difficult to be degraded naturally. In this study, inspired by the “catalytic triad’’ in serine proteases, an enzyme mimic material was developed by combining the proteases’s active sites of serine, histidine and aspartate (S-H-D) with the self-assembling sequence of LKLKLKL and the aromatic group of fluorenylmethyloxycarbonyl (Fmoc). By mixing the monomer of peptides containing separate S, H and D residues with a ratio of 2:1:1, the enzyme mimics were found to co- assemble into nanofibers (Co-HSD) and showed the highest activity towards DEHP degradation because of the synergistic effects of active sites, orderly secondary structure and stable molecular conformation. To further improve ability and applicability, the high active mimetic enzyme was immobilized onto regenerated cellulose (RC) membranes for DEHP degradation in a continuous recycling mode. The RC membranes were first functionalized by the NaIO4 oxidation method to form aldehyde groups and then conjugated with the enzyme mimics via Schiff-base reaction. As a biocatalytic membrane, this membrane could not only effectively degrade DEHP, but also showed good stability, thus establishing a promising biomaterial for large scale biodegradation of DEHP in water decontamination and liquid food depollution.

Efficient biodegradation of DEHP by CM9 consortium and shifts in the bacterial community structure during bioremediation of contaminated soil

Di(2-ethylhexyl) phthalate (DEHP), the most extensively used plasticizer in plastic formulations, is categorized as a priority environmental contaminant with carcinogenic, teratogenic, and mutagenic toxicities. Many isolated microorganisms exhibit outstanding performance as pure cultures in the laboratory but are unable to cope with harsh environmental conditions in the field. In the present study, a microbial consortium (CM9) with efficient functionality was isolated from contaminated farmland soil. CM9 could consistently degrade 94.85% and 100.00% of DEHP (1000 mg/L) within 24 h and 72 h, respectively, a higher efficiency than those of other reported pure and mixed microorganism cultures. The degradation efficiencies of DEHP and di-n-butyl phthalate were significantly higher than those of dimethyl phthalate and diethyl phthalate (p < 0.05). The primary members of the CM9 consortium were identified as Rhodococcus, Niabella, Sphingopyxis, Achromobacter, Tahibacter, and Xenophilus. The degradation pathway was hypothesized to include both de-esterification and β-oxidation. In contaminated soil, bioaugmentation with CM9 and biochar markedly enhanced the DEHP removal rate to 87.53% within 42 d, compared to that observed by the indigenous microbes (49.31%) (p < 0.05). During simulated bioaugmentation, the dominant genera in the CM9 consortium changed significantly over time, indicating their high adaptability to soil conditions and contribution to DEHP degradation. Rhodococcus, Pigmentiphaga and Sphingopyxis sharply decreased, whereas Tahibacter, Terrimonas, Niabella, Unclassified_f_Caulobacteraceae, and Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium showed considerable increases. These results provide a theoretical framework for the development of in situ bioremediation of phthalate (PAE)-contaminated soil by composite microbial inocula.

High-effectively degrade the di-(2-ethylhexyl) phthalate via biochemical system: Resistant bacterial flora and persulfate oxidation activated by BC@Fe3O4

Di-(2-ethylhexyl) phthalate (DEHP) has been classified as a priority pollutant which increased the healthy risk to human and animals dramatically. Hence, a novel biochemical system combined by DEHP-resistant bacterial flora (B) and a green oxidant of persulfate (PS) activated by Nano-Fe3O4 was applied to degrade DEHP in contaminated soil. In this study, the resistant bacterial flora was screened from activated sludge and immobilized by sodium alginate (SAB). Nano-Fe3O4 was coated on biochar (BC@Fe3O4) to prevent agglomerating in soil. X-ray diffraction (XRD) and scanning electron microscope (SEM) were utilized to characterize BC@Fe3O4. Results demonstrated that the treatment of biochemical system (SAB + BC@Fe3O4 + PS) presented the maximum degradation rate about 92.56% within 24 days of incubation and improved soil microecology. The 16S rDNA sequences analysis of soil microorganisms showed a significantly different abundance and a similar diversity among different treatments. Kyoto Encyclopedia of Genes and Genomes (KEGG) functional genes difference analysis showed that some metabolic pathways, such as metabolism of cofactors and vitamins, energy metabolism, cell growth and death, replication and repair, were associated with the biodegradation of DEHP. Besides, DEHP was converted to MEHP and PA by biodegradation, while DEHP was converted to DBP and PA by persulfate and BC@Fe3O4, and then ultimately degraded to CO2 and H2O.

Biodegradability and biodegradation pathway of di-(2-ethylhexyl) phthalate by Burkholderia pyrrocinia B1213

This study was conducted to investigate the biodegradation of di-(2-ethylhexyl) phthalate (DEHP) by Burkholderia pyrrocinia B1213. The results showed that DEHP at concentration of 500 mg/L in a mineral salt medium containing 1.0% yeast extract can be almost completely degraded (98.05%) by strain B1213. The optimal condition for DEHP degradation was pH 7.0, temperature 30 °C. Moreover, B1213 shows better degradation effect on long-chain PAEs, such as DEHP, which provides a great potential for its use in bioremediation of soils contaminated with PAEs. The kinetic studies showed that DEHP depletion curves fit well to the modified Gompertz model. The mono(2-ethylhexyl) phthalate (MEHP), mono-dibutyl phthalate (MBP), phthalic acid (PA) and 4-oxo-hexanoic acid were identified as the metabolites of DEHP by HPLC-ESI-QTOFMS. The detection of MBP and 4-oxo-hexanoic acid as intermediates prompted us to propose a novel and more complete DEHP biodegradation pathway compared to the classic pathway: DEHP is first degraded to MEHP by esterases, which is then converted to MBP through β-oxidation. Then MBP is degraded to PA by esterases, which is then converted to protocatechuate (PCA) under aerobic conditions rapidly. PCA is ultimately cleaved to generate CO2 and H2O via 4-oxo-hexanoic acid.

The complex interactions between novel DEHP-metabolising bacteria and the microbes in agricultural soils

The indigenous microorganisms with the ability of metabolising di-(2-ethylhexyl) phthalate (DEHP) in agricultural soils and their interactions with non-degrading microbes were revealed by DNA-based stable isotope probing coupled with molecular ecological network. Aside from the previously reported DEHP degraders (family Planococcaceae and genus Sphingobacterium), five OTUs representing bacteria affiliated with genus Brevundimona, class Spartobacteria, genus Singulisphaera, genus Dyella and class Ktedonobacteria were linked with DEHP biodegradation. The analysis of the constructed ecological network based on soil microbial communities demonstrated the negative relationships between DEHP degraders and the dominant family Oxalobacteraceae in soils. Additionally, two cultivable bacteria isolated from the same soils, Rhizobium-1 and Ensifer-1, had strong capabilities in degrading DEHP but their involvement in in situ DEHP degradation was questioned, as their DNA was not labelled with 13C from DEHP. These findings provide deeper understanding on the indigenous DEHP-degrading communities and will benefit the remediation of phthalate esters contaminated soils.

Biodegradation patterns of the endocrine disrupting pollutant di(2-ethyl hexyl) phthalate by Fusarium culmorum

Di(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer, which is considered an endocrine disrupting pollutant. Growth kinetics and esterases activity by biochemical tests and polyacrylamide gel electrophoresis were characterized for Fusarium culmorum grown in DEHP-supplemented (1000 mg/L) medium as the only carbon source and in control medium with glucose. Intermediate compounds of biodegraded DEHP were identified by GC-MS. F. culmorum degraded 92% of DEHP within 36 h. DEHP was degraded to butanol, hexanal, catechol and acetic acid. It is suggested that the first two compounds would transform into butanediol and the last two would enter into the Krebs cycle and would be mineralized to CO2 and H2O. DEHP induced eight esterase isoforms, which were different to those constitutive isoforms produced in the control medium. It is suggested that five enzymes (25.7, 29.5, 31.8, 97.6 and 144.5 kDa) detected during the first 36 h be involved in the primary biodegradation of DEHP. The rest of the enzymes (45.9, 66.6 and 202.9 kDa) might be involved in the final steps for DEHP metabolism. F. culmorum has a promising practical application in the treatment of DEHP-contaminated environments because it can secrete specific esterase to breakdown high concentrations of DEHP in a short period of time. This research represents the first approach for the study of esterase involved in the DEHP degradation by fungi using this phthalate as the sole source of carbon and energy.

A novel biodegradation pathway of the endocrine-disruptor di(2-ethyl hexyl) phthalate by Pleurotus ostreatus based on quantum chemical investigation

Di(2-ethyl hexyl) phthalate (DEHP) is a plasticizer that interfere with endocrine systems in mammals. Growth parameters for Pleurotus ostreatus grown on media containing glucose and different concentrations of DEHP (0, 500 and 1000mg/L) were evaluated. The highest biomass production was observed in medium supplemented with 1000mg of DEHP/L. Half-life of DEHP biodegradation, biodegradation constant of DEHP, and percentage of removal efficiency (%E) were also determined. P. ostreatus degraded 100% of DEHP after 504h. %E was 99.3% and 98.4% for 500 and 1000mg of DEHP/L, respectively. Intermediate compounds of biodegraded DEHP were identified by GC-MS and a DEHP biodegradation pathway was proposed using quantum chemical investigation. DEHP might be metabolized through three pathways; a de-esterification pathway, an oxidation pathway and an oxidation-hydrolysis pathway, forming phthalic acid, acetic acid and butanediol, respectively. P. ostreatus degrades and uses (as carbon and energy source) high concentrations of DEHP.

Degradation of di-2-ethylhexyl phthalate (DEHP) by an indigenous isolate Acinetobacter sp. SN13

A bacterial strain capable of degrading di(2-ethylhexyl) phthalate (DEHP) from the artificially contaminated water (with over 90% removal within 5 days of incubation) was isolated from the activated sludge obtained from a regional wastewater treatment plant in Macau Special Administrative Region, China. The isolate was identified as Acinetobacter sp. SN13 by the 16S rRNA gene sequence analysis. Two major experimental parameters, temperature (25, 30, 35 °C) and pH (3–9), were further optimized to enhance the biodegradation efficiency for DEHP. The optimal temperature was 30 °C while there were no significant differences for the pH range tested, 6–9 (p = 0.87). The growth kinetics of this indigenous isolate on DEHP followed the inhibition model, with the maximum degradation rate, the half saturation constant, and the inhibition constant of 124.8 mg l−1 day−1, 272.3 mg l−1, and 720.5 mg l−1, respectively. The inhibition model for the specific growth rate was also simulated using Matlab software, and the respective maximum specific growth rate, half saturation constant, and inhibition constant were 0.1192 day−1, 137.6 mg l−1, and 850.3 mg l−1. The highest degradation rate was achieved when the initial concentration of DEHP was 400 mg l−1. Ferric ion (Fe3+ at 100–1000 μg l−1) showed the stimulatory effect on the DEHP biodegradation, while Mn2+ stimulated the biodegradation at lower concentrations (100 μg l−1) but inhibited at higher concentrations (500–1000 μg l−1). The respective DEHP degradation pathway for the isolate is also proposed by the identification of the following intermediates: mono-(2-ethylhexyl) phthalate (MEHP), phthalic acid (PA), protocatechuate, β-carboxy-cis,cis-muconic acid, and 3-katoadipate.

Effect of bioavailability on the fate of hydrophobic organic compounds and metal in treatment of young landfill leachate by membrane bioreactor

Complex dissolved organic matter (DOM) present in landfill leachate provides reliable media for adsorption of highly hydrophobic contaminants, such as Di 2-ethyl hexyl phthalate (DEHP). In this research, the feasibility of submerged membrane bioreactor (SMBR) for treatment of landfill leachate (LFL) was determined. Later, the operating conditions were optimized for removal of DEHP, COD, NH4+ and PO43−, and finally the effect of bioavailability was examined by introduction of different concentrations of humic acid into the influent. The result revealed that presence of complex agglomerated organic compounds increased the removal efficiency of DEHP and COD, even though DEHP biodegradation rate in sludge dramatically decreased (from 58.8% to 12.8%). MBR retention of different metals in the absence and in the presence of recalcitrant DOM was also studied. Like DEHP, ternary interaction between metals, DOM, and sludge play a pivotal role in their removal efficiency and their concentration in sludge.

Degradation of di(2-ethyl hexyl) phthalate by Fusarium culmorum: Kinetics, enzymatic activities and biodegradation pathway based on quantum chemical modelingpathway based on quantum chemical modeling

Di(2-ethylhexyl) phthalate (DEHP) is a plasticizer widely used in the manufacture of plastics, and it is an environmental contaminant. The specific growth rate (μ), maximum biomass (Xmax), biodegradation constant of DEHP (k), half-life (t1/2) of DEHP biodegradation and removal efficiency of DEHP, esterase and laccase specific activities, and enzymatic yield parameters were evaluated for Fusarium culmorum grown on media containing glucose and different concentrations of DEHP (0, 500 and 1000mg/L). The greatest μ and the largest Xmax occurred in media supplemented with 1000mg of DEHP/L. F. culmorum degraded 95% of the highest amount of DEHP tested (1000mg/L) within 60h of growth. The k and t1/2 were 0.024h−1 and 28h, respectively, for both DEHP concentrations. The removal efficiency of DEHP was 99.8% and 99.9% for 1000 and 500mg/L, respectively. Much higher specific esterase activity than specific laccase activity was observed in all media tested. The compounds of biodegradation of DEHP were identified by GC–MS. A DEHP biodegradation pathway by F. culmorum was proposed on the basis of the intermolecular flow of electrons of the identified intermediate compounds using quantum chemical modeling. DEHP was fully metabolized by F. culmorum with butanediol as the final product. This fungus offers great potential in bioremediation of environments polluted with DEHP.