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The bioflocculant producing bacterial strain – UKD24 was isolated from the domestic sewage treatment plant. The isolated strain was identified as Klebsiella pneumoniae by using 16S rRNA gene sequencing. The K. pneumoniae UKD24 showed remarkable flocculation rates when grown with the carbon sources namely glucose, sucrose and lactose, and many commercial nitrogen sources. Furthermore, the fish waste extract (FE) was used to enhance the productivity of the bioflocculant as a nitrogen supplement and it showed a significant level of flocculation rate similar to the commercial nitrogen sources. The Box–Behnken experiments were designed to predict the optimal conditions for bioflocculant production and it suggested that glucose – 3.247 g L−1, FE – 0.5 g L−1 and inoculum size – 1% are the suitable levels for bioflocculant production. The FTIR analysis of the bioflocculant showed the functional groups related to the polysaccharides and the EEM analysis showed the fluorescence components related to the proteins and humic acids. The biochemical composition of the bioflocculant was identified as polysaccharides (24.36 ± 1.5%) and protein (12.15 ± 0.2%). The tested optimum conditions of the bioflocculant to induce flocculation were tested in the kaolin wastewater and it showed that the optimum dosage of the flocculant was 5 mg L−1 and the pH range was broad as 5–10. The cation dependency tests revealed that the monovalent and divalent cations are highly suitable for flocculation while the trivalent cations showed moderate flocculation. The Cr(VI) removal efficiency of the bioflocculant showed that ∼35% of heavy metal is trapped into flocks during the flocculation.

The aqueous extract of various plants like Coriandrum sativum (AECS), Alternanthera tenella colla (AEAT), Spermacoce hispida (AESH) and Mollugo verticillata (AEMV) was studied for its hexavalent chromium (CrVI) reduction property. Even though antioxidant activity was present, AEAT, AESH and AEMV did not reduce CrVI. AECS showed rapid and dose-dependent CrVI reduction. The efficient reduction of 50 mg/L of CrVI using AECS was attained in the presence of 250 µg/mL of starting plant material, incubating the reaction mixture at pH 2, 30°C and agitation at 190 rpm. Under such conditions, about 40 mg/L of CrVI was reduced at 3 h of incubation. FT-IR analysis revealed the involvement of phenols, alcohols, alpha-hydroxy acid and flavonoids present in the AECS for the CrVI reduction. These results indicate that not all the plant extracts with rich antioxidants are capable of reducing CrVI. Using the conditions standardized in the present study, AECS reduced about 80% of CrVI present in the tannery effluent...

The main concern of extensive production of biobutanol has been associated with the high cost of the substrate and the relatively low tolerance of Clostridia to biobutanol production. In this study, the use of fermentable cassava waste residue (CWR) as substrate for biobutanol production was investigated using solvent-tolerant Clostridium sp. Four of obligatory, solvent-producing bacteria were isolated from sago industry waste sites. The NSW, PNAS1, SB5 and SBI4 strains showed identical profiles of 16S rRNA gene sequence similarity of Bacillus coagulans, Clostridium bifermentans and Clostridium sp. (97% similarity) and a wide range of carbohydrate substrate; however, the CWR was found to be suitable for the production of biobutanol considerably. Batch culture study was carried out using parameters such as time and temperature and carbon sources have been studied and optimized. Using pre-optimized CWR medium, significant amount of solvent production was observed in NSW, PNAS1, SB5 and SBI4 with 1.53, 3.36, 1.56 and 2.5 g L

The continuous release of toxic persistent organic pollutants (POPs) into the environment has raised a need for effective cleanup methods. The tremendous natural diversity of microbial catabolic mechanisms suggests that catabolic routes may be applied to the remediation of POP-contaminated fields. A large number of the recalcitrant xenobiotics have been shown to be removable via the natural catabolic mechanisms of microbes, and detailed biochemical studies of the catabolic methods, together with the development of sophisticated genetic engineering, have led to the use of synthetic microbes for the bioremediation of POPs. However, the steric effects of substituted halogen moieties, microbe toxicity, and the low bioavailability of POPs still deteriorate the efficiency of removal strategies based on natural and synthetic catabolic mechanisms. Recently, abiotic redox processes that induce rapid reductive dehalogenation, hydroxyl radical-based oxidation, or electron shuttling have been reasonably coupled with microbial catabolic actions, thereby compensating for the drawbacks of biotic processes in POP removal. In this review, we first compare the pros and cons of individual methodologies (i.e., the natural and synthetic catabolism of microbes and the abiotic processes involving zero-valent irons, advanced oxidation processes, and small organic stimulants) for POP removal. We then highlight recent trends in coupling the biotic–abiotic methodologies and discuss how the processes are both feasible and superior to individual methodologies for POP cleanup. Cost-effective and environmentally sustainable abiotic redox actions could enhance the microbial bioremediation potential for POPs.

The dioxin isomer 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TeCDD) has been reported as the deadliest compound known to science. Due to its highly recalcitrant nature and low bioavailability, it is stubborn toward bioremediation and chemical treatment. Efforts to degrade it using one single technique have not accomplished the desired results. In this study, we have tried to develop an integrated 2,3,7,8-TeCDD removal process using palladized iron nanoparticles (Pd/nFe) for initial reductive dechlorination under anoxic conditions and subsequent oxidative biomineralization. Using laboratory synthesized Pd/nFe, 2,3,7,8-TeCDD was completely dechlorinated to form the end product dibenzo-p-dioxin (DD). Oxidative degradation of DD was successfully achieved by growing active cells of a dioxin-degrading microorganism Sphingomonas wittichii RW1 (DSM 6014) under aerobic culture conditions. Metabolite identification was done by high performance liquid chromatography (HPLC) and whole cell protein was measured as the indicator for cell growth. To the best of our knowledge, this is the first report on integrated hybrid degradation method for 2,3,7,8-TeCDD.

BACKGROUND: Polybrominated diphenyl ethers (PBDEs) are emerging persistent organic pollutants. Degradation of PBDEs is a significant challenge owing to their extreme persistence and recalcitrance nature. The objective of this study was to evaluate the effect of a sequential nano-bio treatment using nanoscale zero-valent iron (nZVI) and diphenyl ether degrading bacteria Sphingomonas sp. PH-07 for degradation of PBDEs. RESULTS: In the bacterial tolerance test for determining the maximum endurable concentration of nZVI, the PH-07 strain was able to grow in the presence of nZVI up to 5 g L−1 in minimal salt medium by using non-brominated diphenyl ether as growth substrate. Reductive debromination of decabrominated diphenyl ether (deca-BDE; 1 mg) with nZVI (100 mg per vial) resulted in a 67% reduction of deca-BDE and produced various intermediates ranging from nona-BDEs to tri-BDEs during a 20 day period. Additional experiments with 2,4,4′-tri-BDE and 2,4,6-tri-BDE as initial substrates revealed that both tri-BDEs were further debrominated to mono-BDEs. Following the reductive debromination process, reaction mixtures were aerobically treated with DE-grown Sphingomonas sp. PH-07 strain to mineralize the low brominated-DEs (tri-BDEs—mono-BDEs) for additional 4 days. During bacterial treatment, the low brominated-DEs were biologically degraded to bromophenols and other prospective metabolites. CONCLUSIONS: The nZVI-biological sequential treatment method was found to be effective for degradation of PBDEs through reductive debromination followed by biological oxidation. This hybrid treatment method may lead to the development of a remediation strategy for highly halogenated environmental pollutants in contaminated sites. Copyright © 2011 Society of Chemical Industry

The well-known bacterium Sphingomonas wittichii RW1 catabolically degrades dibenzo-p-dioxin and dibenzofuran, as well as their chlorinated derivatives. The catabolic degradation of dioxin is initiated by a ring-hydroxylating dioxygenase. The biotransformation of carbazole by S. wittichii RW1 was determined in the present study. Dioxin dioxygenase from the dibenzofuran induced RW1 strain was thought to be unable to attack carbazole, which includes a heterocyclic aromatic dibenzopyrrole system. However, our results showed that carbazole was transformed to anthranilic acid and catechol. The color of the culture suspension changed upon addition of carbazole due to formation of a nitrogen-containing metabolite. Relevant metabolic intermediates were identified by gas chromatographic mass spectrometry and electrospray ionization time-of-flight mass spectrometry with comparison to the corresponding authentic compounds. The dioxygenase of the dibenzofuran induced RW1 attacked at the angular position adjacent to the nitrogen atom to give a dihydroxylated metabolic intermediate. Contrary to predictions made in previous reports, S. wittichii RW1 displayed positive catabolic activity toward carbazole.

A new bacterial strain PH-06 was isolated using enrichment culture technique from river sediment contaminated with 1,4-dioxane, and identified as belonging to the genus Mycobacterium based on 16S rRNA sequencing (Accession No. EU239889). The isolated strain effectively utilized 1,4-dioxane as a sole carbon and energy source and was able to degrade 900 mg/l 1,4-dioxane in minimal salts medium within 15 days. The key degradation products identified were 1,4-dioxane-2-ol and ethylene glycol, produced by monooxygenation. Degradation of 1,4-dioxane and concomitant formation of metabolites were demonstrated by GC/MS analysis using deuterium labeled 1,4-dioxane (1,4-dioxane-d8). In addition to 1,4-dioxane, this bacterium could also transform structural analogues such as 1,3-dioxane, cyclohexane and tetrahydrofuran when pre-grown with 1,4-dioxane as the sole growth substrate. Our results suggest that PH-06 can maintain sustained growth on 1,4-dioxane without any other carbon sources.

Several aerobic bacteria possess unique catabolic pathways enabling them to degrade persistent organic pollutants (POPs), including polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs), polybrominated diphenylethers (PBDEs), and polychlorinated biphenyls (PCBs). The catabolic activity of aerobic bacteria employed for removal of POPs in the environment may be modulated by several biotic (i.e. fungi, plants, algae, earthworms, and other bacteria) and abiotic (i.e. zero-valent iron, advanced oxidation, and electricity) agents. This review describes the basic biochemistry of the aerobic bacterial catabolism of selected POPs and discusses how biotic and abiotic agents enhance or inhibit the process. Solutions allowing biotic and abiotic agents to exert physical and chemical assistance to aerobic bacterial catabolism of POPs are also discussed.

The dioxin-degrading strain Pseudomonas veronii PH-03 was isolated from contaminated soil by selective enrichment techniques. Strain PH-03 grew on dibenzo-p-dioxin and dibenzofuran as a sole carbon source. Further, 1-chlorodibenzo-p-dioxin, 2-chlorodibenzo-p-dioxin and other dioxin metabolites, salicylic acid, and catechol were also metabolized well. Resting cells of strain PH-03 transformed dibenzo-p-dioxin, dibenzofuran, 2,2',3-trihydroxybiphenyl, and some chlorodioxins to their corresponding metabolic intermediates such as catechol, salicylic acid, 2-hydroxy-(2-hydroxyphenoxy)-6-oxo-2,4-hexadienoic acid, and chlorocatechols. The formation of these metabolites was confirmed by comparison of gas chromatography-mass spectrometry (GC-MS) data with those of authentic compounds. Although we did observe the production of 3,4,5,6-tetrachlorocatechol (3,4,5,6-TECC) from 1,2,3,4-tetrachlorodibenzo-p-dioxin (1,2,3,4-TCDD) with resting cell suspensions of PH-03, growth of strain PH-03 in the presence of 1,2,3,4-TCDD was poor. This result suggests that strain PH-03 is unable to utilize 3,4,5,6-TECC, even at very low concentration (0.01 mM) due to its toxicity. In cell-free extracts of DF-grown cells, 2,2',3-trihydroxybiphenyl dioxygenase, 2-hydroxy-6-oxo-6-phenyl-2,4-hexadienoic acid hydrolase, and catechol-2,3-dioxygense activities were detected. Moreover, the activities of meta-pyrocatechase and 2,2',3-trihydroxybiphenyl dioxygenase from the crude cell-free extracts were inhibited by 3-chlorocatechol. However, no inhibition was observed in intact cells when 3-chlorocatechol was formed as intermediate.

This study investigated the cytotoxicity, genotoxicity, and growth inhibition effects of four different inorganic nanoparticles (NPs) such as aluminum (nAl), iron (nFe), nickel (nNi), and zinc (nZn) on a dibenzofuran (DF) degrading bacterium Agrobacterium sp. PH-08. NP (0–1,000 mg L−1) -treated bacterial cells were assessed for cytotoxicity, genotoxicity, growth and biodegradation activities at biochemical and molecular levels. In an aqueous system, the bacterial cells treated with nAl, nZn and nNi at 500 mg L−1 showed significant reduction in cell viability (30–93.6 %, p

Laccase was produced from Pleurotus florida under solid-state fermentation, and the production was optimized by response surface methodology. The predicted maximum laccase production of 8.81 U g(-1) was obtained by the optimum concentration of malt extract, banana peel, wheat bran and CuSO4, which was found to be 0.69 g, 10.61 g, 10.68 g and 77.15 ppm, respectively. The validation results suggested that the laccase production was 7.96 U g(-1) in the optimized medium, which was close to the predicted value. Decolorization efficiency of P. florida laccase was evaluated against malachite green (MG). Rapid decolorization of MG dye was observed, and a dark-coloured precipitate was formed in the reaction mixture. HPLC analysis indicated that the laccase enzyme degraded MG by the demethylation process. The toxicity of MG was reduced to 67% after the treatment with laccase, which was confirmed by a phytotoxicity study.

In this work, the influence of different metal ions on laccase activity and laccase-catalyzed dye decolorization was investigated under in vitro conditions using crude laccase obtained from a white rot fungus Ganoderma lucidum. Laccase activity was enhanced by metal ions such as Ca(2+), Co(2+), Cu(2+) and Zn(2+) at low concentrations (1mM). Increasing the concentration of metal ions except that of Cu(2+) and Zn(2+) up to 5mM and above decreased the enzyme activity. Among several heavy metals, Fe(2+) highly inhibited the enzyme activity. Effect of metal ions was tested on decolorization of two reactive dyes, namely Remazol black-B (RB-5) and Remazol brilliant blue R (RBBR) at a concentration of 50 mg l(-1). The presence of heavy metals generally did not exert much influence on the decolorization except Fe(2+). Cu(2+) and Cr(6+) enhanced the decolorization of both dyes. In the presence of 1mM Cu(2+), 94% of RB-5 and 35.5% of RBBR were decolorized during 1h incubation. G. lucidum laccase was able to tolerate mixture of several metal ions. Treatment of simulated reactive dye effluent by laccase showed that the redox mediator system is necessary for effluent decolorization. Syringaldehyde, a natural redox mediator, was very effective than the synthetic mediator 1-hydroxybenzotriazole (HBT). The initial rate of effluent decolorization in presence of syringaldehyde (0.0831 h(-1)) was 5.6 times higher than HBT (0.0152 h(-1)). Although the rate of decolorization was markedly decreased in the effluent containing mixed metal ions, presence of syringaldehyde showed effective decolorization. This study indicates that G. lucidum laccase and natural redox mediator system could be a potential candidate for color removal from reactive dye effluent.

Polybrominated diphenyl ethers (PBDEs) are common flame-retardant chemicals that are used in diverse commercial products such as textiles, circuit boards, and plastics. Because of the widespread production and improper disposal of materials that contain PBDEs, there has been an increasing accumulation of these compounds in the environment. The toxicity and bioavailability of PBDEs are variable for different congeners, with some congeners showing dioxin-like activities and estrogenicity. The diphenyl ether-utilizing bacterium Sphingomonas sp. PH-07 was enriched from activated sludge of a wastewater treatment plant. In liquid cultures, this strain mineralized 1 g of diphenyl ether per liter completely within 6 days. The metabolites detected and identified by gas chromatography/mass spectrometry (MS) and electrospray ionization/MS analysis corresponded with a feasible degradative pathway. However, the strain PH-07 even catabolized several brominated congeners such as mono-, di-, and tribrominated diphenyl ethers thereby producing the corresponding metabolites.

Removal of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) from fly ash poses a serious problem. In the study presented here, we used a microbial biocatalyst which is a mixture of 4 bacterial and 5 fungal dioxin-degrading strains. The ability of this biocatalyst to bioremediate PCDD/Fs from contaminated municipal solid waste incinerator (MSWI) fly ash was examined by solid-state fermentation under laboratory conditions. Treatment of MSWI fly ash with the microbial biocatalyst for 21 days resulted in a 68.7% reduction in total toxic PCDD/Fs. Further analyses revealed that the microbial biocatalyst also removed 66.8% of the 2,3,7,8-substituted congeners from the fly ash. During the treatment period, the presence of the individual strains composing the microbial biocatalyst was monitored by the amplification of strain-specific DNA sequences followed by denaturing gradient gel electrophoresis (DGGE). This analysis showed that all of the bacterial and fungal strains composing this dioxin-degrading microbial mixture maintained under the dioxin treatment conditions. These results demonstrate that this microbial biocatalyst could potentially be used in the bioremediation of PCDD/Fs from contaminated fly ash.

The ability of Sphingomonas wittichii strain RW1 to remove polychlorinated dibenzo-p-dioxins (PCDDs) from fly ash was investigated. All experiments were carried out in a slurry-phase system. Preliminary studies with resting cells of strain RW1 in a model fly ash system showed the complete removal of dibenzofuran (DF) and 81% of dibenzo-p-dioxin (DD). Incubation of real fly ash collected from municipal waste incinerators with strain RW1 for 15 days resulted in a 75.5% reduction in toxic PCDDs. When the same experiment was carried out using dead strain RW1 cells a 20.2% reduction in toxic PCDDs was observed, indicating that adsorption onto biomass was an important factor in dioxin elimination. Further analyses revealed that live strain RW1 cells removed 83.8% of the 2,3,7,8-substituted congeners from the fly ash, while dead cells removed 32.1% of the same congeners. To enhance the removal efficiency of toxic PCDDs, the effects of adding surfactant, repeated inoculation, and pre-adaptation of cultures were also studied. The removal of toxic PCDDs was enhanced by up to 10.3% upon repeated inoculation of the strain RW1, but was not much affected by the addition of surfactant. The present results suggest that S. wittichii strain RW1 is a potential candidate for the industrial removal of PCDDs from incinerator fly ash.

Firstly, we would like to thank to Halden (Halden, 2006) for his interest in, and kind comments on, our recent article (Nam et al. 2005). Removal of polychlorinated dibenzo-pdioxins (PCDDs) from fly ash is a serious problem and being investigated intensively to develop an eco-friendly treatment method. The gram negative bacterium Sphingomonas wittichii strain RW1 has been shown its versatility in degradation of low chlorinated dioxins in different matrices such as liquid and soil environments (Wittich et al. 1992; Wilkes et al. 1996; Halden et al. 1999). Recent studies proved that its biotransformation