Decolorization of textile dyes by laccases from a newly isolated strain of Trametes modesta

Four ligninolytic fungi, Trametes modesta, Trametes hirsuta, Trametes versicolor and Sclerotium rolfsii, were compared for their ability to produce laccases. The fungal laccases were screened for their ability to decolorize eight synthetic dyes (anthraquinone, azo, indigo and triarylmethane). The decolorization rate depended both on the source of the enzyme preparation and on the structure of the dye. Based on laccase production and dye decolorizing ability, T. modesta was selected for further studies. All the tested dyes were decolorized by the T. modesta laccase most efficiently under acid conditions (pH 3-6) but the optimum pH for decolorization of the individual dye varied. The decolorization rate of this laccase increased with the rise in temperature to 50-60 degrees C. The decolorization efficiency of T. modesta laccase was improved remarkably in the presence of mediators like 1-hydroxybenzotriazole and 2-methoxyphenothiazine.     

Optimal decolorization and kinetic modeling of synthetic dyes by Pseudomonas strains

Pseudomonas spp were isolated from an anaerobic-aerobic dyeing house wastewater treatment facility as the most active azo-dye degraders. Decolorization of azo dyes and non-azo dyes including anthraquinone, metal complex and indigo was compared with individual strains and a bacterial consortium consisting of the individual strain and municipal sludge (50 50wt). The consortium showed a significant improvement on decolorization of two recalcitrant non-azo dyes, but little effect on the dyes that the individual strains could degrade to a great or moderate extent. Decolorization of Acid violet 7 (monoazo) by a Pseudomonas strain GM3 was studied in detail under various conditions. The optimum decolorization activity was observed in a narrow pH range (7-8), a narrow temperature range (35-40 degrees C), and at the presence of organic and ammonium nitrogen. Nitrate had a severe inhibitory effect on azo dye decolorization: 10 mg/L led to 50% drop in decolorization activity and 1000 mg/L to complete activity depression. A kinetic model is established giving the dependence of decolorization rate on cell mass concentration (first-order) and dye concentration (half order). The rate increased with temperature from 10 to 35 C, which can be predicted by Arrhenius equation with the activation energy of 16.87 kcal/mol and the frequency factor of 1.49 x 10(11) (mg L)1/2/g DCM min.     

Investigation of isolation and immobilization of a microbial consortium for decoloring of azo dye 4BS

Eight high-effective decolorization strains were isolated by enrichment using Direct Fast Scarlet 4BS as sole source of carbon and energy. The optimal microbial consortium consisting of fungus 8-4(*) and bacterium 1-10 was selected by optimizing combination decolorization experiments with these eight freely suspended strains, whose decolorization activity was higher than individual strains due to synergistic reaction with each other. The optimal microbial consortium was also immobilized using polyvinyl alcohol (PVA) as the carrier. This paper optimized the immobilization and operational conditions, investigated the effect of the environmental factors (temperature, pH and dissolved oxygen (DO)) and initial dye concentration on the rate of decolorization by immobilized microbial consortium. The results showed that the optimal decolorization activity was observed in pH range (5-8), temperature range (25-40 degrees C) under shaking culture of high DO level. Decolorization with the optimal microbial consortium gave a relatively high maximum decolorization activity (ca. 81.25 mgl(-1)h(-1)), which occurred at a dye concentration of 1000 mgl(-1), suggesting the applicability of the strains in remediation of wastewater containing high azo dye concentrations. The immobilized beads could be reused for more than 30 cycles, without losing any degradation capacity. The changes of UV-visible spectra and the change curve of COD of 4BS solution before and after decolorization cultivation and the proliferation and distribution of microbial consortium in gel beads were also microscopically observed, which could be used for conferring the decolorization mechanisms of dye 4BS.     

Kinetic characteristics of bacterial azo-dye decolorization by Pseudomonas luteola

A Pseudomonas luteola strain expressing azoreductase activity was utilized to remove the color of an azo dye (reactive red 22) from contaminated solutions. The effects of substrate concentrations, medium compositions, and operation parameters (e.g., pH, temperature, dissolved oxygen, etc.) on decolorization of the azo dye by a P. luteola strain were systematically investigated to reveal the key factors that dominate the performance of azo-dye decolorization. The metabolites resulting from bacterial decolorization were analyzed by high-performance liquid chromatography (HPLC) and mass spectrometery (MS). The results show that the dissolved oxygen and glucose concentration retarded decolorization of reactive red 22 by P. luteola. The optimal azo-dye decolorization occurred at 37 degrees C, while more rapid decolorization took place over pH 7-9. Yeast extract and tryptone strongly enhanced the decolorization. The Michaelis-Menten model can satisfactorily describe the dependence of specific decolorization rate on the concentration of substrate (reactive red 22 or yeast extract). Decolorization of the azo dye by intact cells of P. luteola was essentially independent of the growth phase, whereas the azoreductase activity of the cell-free extract decreased in the order of late-stationary phase > early-stationary phase > mid-log phase. This suggests that mass transfer of the azo dye across the cell membrane may be the rate-limiting step. The HPLC and MS analyses suggest that both partial reduction and complete cleavage of the azo bond could contribute to decolorization of reactive red 22 by P. luteola.     

Laccase-membrane reactors for decolorization of an acid azo dye in aqueous phase: Process optimization

In the present investigation, performance of various laccase-membrane reactor configurations including direct enzyme contact, enzyme impregnated, immobilized enzyme and a reactor system based on laccase immobilization in chitosan membranes for decolorization of azo dye (acid black 10 BX) were examined using laccase enzyme purified from white rot fungi Pleurotus ostreatus 1804. A five-step laccase purification procedure was employed, which improved the enzymatic activity by 8.27 folds. Laccase was confirmed by comparing with the standard marker using SDS-PAGE electrophoresis, which showed molecular weight of 63 kDa. Experimental data showed that laccase has great potential for color removal without addition of external redox mediators. Various process parameters viz. aqueous phase of pH 6.0, enzyme concentration of 1.75 U/ml, dye concentration of 20 mg/L, temperature of 30 °C and reaction time of 120 min were optimized to achieve maximum decolorization efficiencies. Moreover, different laccase-membrane reactor configurations were tested to determine the efficacy of repeated application of laccase on dye decolorization process. Among the different reactor configurations employed, laccase encapsulated in chitosan membrane showed advantages such as short-term contact period and reusability of enzyme for a number of cycles.     

Combined anaerobic-aerobic treatment of azo dyes--a short review of bioreactor studies

The most logical concept for the removal of azo dyes in biological wastewater treatment systems is based on anaerobic treatment, for the reductive cleavage of the dyes' azo linkages, in combination with aerobic treatment, for the degradation of the products from azo dye cleavage, aromatic amines. Since the 1990s, several research papers have been published on combined, sequential or integrated, anaerobic-aerobic bioreactor treatment of azo dye-containing wastewater. The extent of azo dye reduction in the anaerobic phase of those bioreactor systems was generally high, albeit the process often required long reaction times, a limitation that can easily be remedied by making use of the property of redox mediators to speed up the process. The consequent removal of aromatic amines under aerobic conditions was less unequivocal. Although analytical data indicate that many of the aromatic amines were removed from the wastewater, and although the limited amount of available toxicity data all show far-reaching detoxification during aerobic treatment, it is clear that not all aromatic amines can be completely mineralized.     

Decolorization and toxicity of reactive anthraquinone textile dyes under methanogenic conditions

Reductive decolorization of two anthraquinone reactive dyes (Reactive Blue 4, RB4; Reactive Blue 19, RB19) under methanogenic conditions was performed using a mixed, methanogenic culture. Decolorization of the two anthraquinone dyes was investigated to evaluate the rate and extent of color removal as well as to assess possible toxic effects of the dyes and their decolorization product(s) on the methanogenic culture as a function of initial dye concentration ranging from 50 to 300 mg x L(-1). A dextrin/peptone mixture was used as the carbon and electron source. A high rate and extent of color removal was achieved ranging from 4.3 to 29.9 mg x L(-1)h(-1) and 73-91% for RB4, and 13.0-74.4 mg x L(-1)h(-1) and 90-95% for RB19. Initial RB4 concentrations up to 100 mg x L(-1) did not result in any significant inhibition. Both the 200 and 300 mg x L(-1) RB4-amended cultures, and all RB19-amended cultures resulted in severe inhibition of both acidogenesis and methanogenesis. Sequential dye addition at 300 mg x L(-1) for both RB4 and RB19 resulted in accumulation of volatile fatty acids (VFAs) and a very low methane production at the end of the first dye addition after 44 days of incubation. However, at the end of the second dye addition, after a relatively long incubation (384 days), recovery of methanogens in the RB4-amended culture was observed in contrast to the complete inhibition of methanogenesis in the RB19-amended culture. Therefore, RB19 resulted in a higher degree of inhibition of both acidogenesis and methanogenesis than RB4. Addition of dextrin/peptone to dye-inhibited cultures resulted in acidogenesis and a gradual recovery of methanogenesis (mainly aceticlastic methanogenesis) in the RB4-inhibited culture, and a slow recovery of acidogenesis but no recovery of methanogenesis in the RB19-inhibited culture. In contrast, addition of 80% H(2)-20% CO(2) gas to dye-inhibited cultures resulted in recovery of hydrogenotrophic methanogenesis in both the RB4- and RB19-inhibited cultures. In spite of the relatively severe inhibition of the two anthraquinone dyes on the mixed, methanogenic culture, a high extent of color removal was achieved.     

Application of the laccase,produced on coconut flesh by Pleurotus florida for dye decolorization

We investigated the ability of Pleurotus florida to produce laccase on coconut flesh as a solid substrate fermentation. The decolorization of two structurally different dyes such as azo (Reactive Blue 198) and triphenylmethane dye (Malachite Green) were analysed. The decolorization of Reactive blue 198 and Malachite Green at 8 hrs was 93% and 63% respectiely. The untreated and treated dye was characterized by UV-Vis spectral and fourier transform infrared (FTIR) Spectroscopy scan. FTIR analysis pointed out the involvement of alkene (C=C) and carboxylic (C-O) groups in the decolorization process. The toxicity with respect to Allium cepa root inhibition was measured to demonstrate the potential of laccase in the detoxication and bioremediation process.     

Further treatment of decolorization liquid of azo dye coupled with increased power production using microbial fuel cell equipped with an aerobic biocathode

A microbial fuel cell (MFC) incorporating a recently developed aerobic biocathode is designed and demonstrated. The aerobic biocathode MFC is able to further treat the liquid containing decolorization products of active brilliant red X-3B (ABRX3), a respective azo dye, and also provides increased power production. Batch test results showed that 24.8% of COD was removed from the decolorization liquid of ABRX3 (DL) by the biocathode within 12 h. Metabolism-dependent biodegradation of aniline-like compound might be mainly responsible for the decrease of overall COD. Glucose is not necessary in this process and contributes little to the COD removal of the DL. The similar COD removal rate observed under closed circuit condition (500 Ω) and opened circuit condition indicated that the current had an insignificant effect on the degradation of the DL. Addition of the DL to the biocathode resulted in an almost 150% increase in open cycle potential (OCP) of the cathode accompanied by a 73% increase in stable voltage output from 0.33 V to 0.57 V and a 300% increase in maximum power density from 50.74 mW/m² to 213.93 mW/m². Cyclic voltammetry indicated that the decolorization products of the ABRX3 contained in the DL play a role as redox mediator for facilitating electron transfer from the cathode to the oxygen. This study demonstrated for the first time that MFC equipped with an aerobic biocathode can be successfully applied to further treatment of effluent from an anaerobic system used to decolorize azo dye, providing both cost savings and high power output.     

纳米TiO2-AC负载膜光催化降解偶氮染料

以钛酸四丁酯和活性炭为主要原料，采用溶胶-凝胶-浸渍法制备纳米TiO2-AC负载膜催化剂，并借助流化床反应器分别对两种典型的偶氮类染料橙黄C、活性艳红X-3B模拟废水进行了UV光降解、暗态吸附及光催化降解研究，探讨了pH值、外加氧化剂对光催化去除率的影响，同时对催化剂进行回收再生利用试验。结果表明：纳米TiO2-AC负载膜具有很好的光催化活性、吸附特性及可再生性，两种染料1h的光催化降解率最高分别可达99．71％和97．12％。反应过程中固、液、气三相能够有效分离，实现了反应与分离的一体化。     

Comparative studies on potential of consortium and constituent pure bacterial isolates to decolorize azo dyes

The decolorization potential of the consortium HM-4 constituted by mixing four laboratory isolates identified as Bacillus cereus (BN-7), Pseudomonas putida (BN-4), Pseudomonas fluorescens (BN-5) and Stenotrophomonas acidaminiphila (BN-3) was compared with that of individual isolates. Six different azo dyes viz., C.I. Acid Red 88 (AR-88), C.I. Acid Red 119 (AR-119), C.I. Acid Red 97 (AR-97), C.I. Reactive Red 120 (RR-120), C.I. Acid Blue 113 (AB-113) and C.I. Acid Brown 100 (AB-100) were used in this study. The individual bacterial isolates were not able to completely decolorize these dyes, except for dyes AR-119 and AB-113. The consortium HM-4 was able to decolorize all the dyes used at an initial dye concentration of 20 mg L⁻¹ at a significantly higher rate as compared to that achieved by individual isolates.     

Enhanced transformation of triclosan by laccase in the presence of redox mediators

Triclosan (TCS), an antimicrobial agent, is an emerging and persistent environmental pollutant that is often found as a contaminant in surface waters and sediments; hence, knowledge of its degradability is important. In this study we investigated laccase-mediated TCS transformation and detoxification, using laccase (from the fungus Ganoderma lucidum) in the presence and absence of redox mediators. Transformation products were identified using HPLC, ESI-MS and GC-MS, and transformation mechanisms were proposed. In the absence of redox mediator, 56.5% TCS removal was observed within 24 h, concomitant with formation of new products with molecular weights greater than that of TCS. These products were dimers and trimers of TCS, as confirmed by ESI-MS analysis. Among the various mediators tested, 1-hydroxybenzotriazole (HBT) and syringaldehyde (SYD) significantly enhanced TCS transformation (∼90%). The presence of these mediators resulted in products with lower molecular weights than TCS, including 2,4-dichlorophenol (2,4-DCP; confirmed by GC-MS) and dechlorinated forms of 2,4-DCP. When SYD was used as the mediator, dechlorination resulted in 2-chlorohydroquinone (2-CHQ). Bacterial growth inhibition studies revealed that laccase-mediated transformation of TCS effectively decreased its toxicity, with ultimate conversion to less toxic or nontoxic products. Our results confirmed the involvement of two mechanisms of laccase-catalyzed TCS removal: (i) oligomerization in the absence of redox mediators, and (ii) ether bond cleavage followed by dechlorination in the presence of redox mediators. These results suggest that laccase in combination with natural redox mediator systems may be a useful strategy for the detoxification and elimination of TCS from aqueous systems.     

Removal of azo and anthraquinone dyes from aqueous solutions by Eichhornia Crassipes

The rate of adsorption of two azo and four anthraquinone anionic dyes on Eichhornia Crassipes (E.C.) has been studied. Raw E.C. and three aminated derivatives of E.C. with different nitrogen percent were used as dye adsorbents. The parameters studied include the amount of substrate, shaking time, chemical structure, concentration of dyestuff and pH of dyeing bath. Simple kinetic adsorption models of dynamics and adsorption parameters for the Langmuir and Freundlich isotherms were determined. A higher nitrogen percent of aminated E.C. showed a higher adsorption capacity than other derivatives. The kinetic adsorption models indicate that the decolourization was complete in a relatively short time (10 min) and the reaction taking place is of the first order. The equilibrium data fit well with the Freundlich model of adsorption for the six dyes. Only dye IV (C.I.A Acid Blue 25) conform both Freundlich and Langmuir adsorption isotherms.     

The photocatalytic degradation of reactive black 5 using TiO2/UV in an annular photoreactor

The textile effluent is a major industrial polluter because it is highly colored, containing about 15% unfixed dyes as well as high levels of salts that can potentially be discharged into the environment. Photocatalytic oxidation using an thin gap annular UV reactor with TiO2 was used to break down the colour of a synthetic effluent ranging up to 400 ppm in dye concentration of Reactive Black 5 and up to 80 g/L in NaCl. Results show that the reaction kinetics was dominated by the TiO2 loading, the initial dye concentration, and the dissolved oxygen concentration; with the other parameters showing less significant effects. High rates of decolorization were found, with a linear fit to the Langmuir-Hinshelwood equation yielding a reaction rate constant (k) of 2.45 ppm/min, and an adsorption equilibrium constant (K) of 0.048 ppm(-1) based on color removal. The presence of the combination of high dissolved oxygen (15 ppm) and sodium chloride (up to 80 g/L) was found to enhance the decolorization and mineralization rates of the reactive dye. However, pH was found to not significantly affect the degradation rate. Since textile effluent is strongly alkaline, this result is significant, as no solution neutralisation is required and direct treatment of the effluent is possible.     

Anaerobic/aerobic treatment of coloured textile effluents using sequencing batch reactors

Conventional biological wastewater treatment plants do not easily degrade the dyes and polyvinyl alcohols (PVOH) in textile effluents. Results are reported on the possible advantages of anaerobic/aerobic cometabolism in sequenced redox reactors. A six phase anaerobic/aerobic sequencing laboratory scale batch reactor was developed to treat a synthetic textile effluent. The wastewater included PVOH from desizing and an azo dye (Remazol Black). The reactor removed 66% of the applied total organic carbon (load F: M 0.15) compared to 76% from a control reactor without dye. Colour removal was 94% but dye metabolites caused reactor instability. Aromatic amines from the anaerobic breakdown of the azo dyes were not completely mineralised by the aerobic phase. Breakdown of PVOH by the reactor (20-30%) was not as good as previous reports with entirely aerobic cultures. The anaerobic cultures were able to tolerate the oxygen and methane continued to be produced but there was a deterioration in settlement.     

Decolorization of an azo dye by unacclimated activated sludge under anaerobic conditions

Studies on the reductive decolorization of a complex azo dye, Reactive Red 3.1, were made as part of the development of a practical approach to better exploit the metabolic potential of biomass in wastewater treatment. Decolorization was achieved at low and variable rates by mixed microbial cultures under various environmental conditions, including low pH and high salt concentration. It was caused by reductive cleavage of the azo bond to yield two aromatic amines. More reliable and effective decolorization rates, of up to 20–30 mg l⁻¹ h⁻¹, were given by unadapted activated sludge, (6 g l⁻¹) incubated with 400 mg l⁻¹ of Reactive Red 3.1 under anaerobic conditions. Decolorization also occurred best in static conditions.     

The Simultaneous Bioremoval of Cr(III) and Dye by Immobilized <Emphasis Type="Italic">Phanerochaete Chrysosporium</Emphasis>

To attain better removal efficiency and higher toxic resistance, the alginate was used to immobilize Phanerochaete chrysosporium BKM-F-1767 in this study. And according to the characteristics of tannery wastewater, inhibitory effect of Cr(III) to the decolorization was investigated and adsorption kinetics of Cr(III) by the immobilized P. chrysosporium had been established. Furthermore, the Acid Violet 7 and Basic Fuchsin contributed as the experimental dyes in the paper, the removal studies were performed at an initial pH of 4.5. The combined effects of Cr and dyes on the simultaneous removal properties were determined in a batch system at different levels of Cr and dyes. Moreover, the dose-response relationship and a kinetic equation describing the simultaneous removal properties had been established. The results have proved that the immobilized P. chrysosporium has the ability to treat the tannery wastewater.     

Metabolism of azo dyes by Lactobacillus casei TISTR 1500 and effects of various factors on decolorization

Lactobacillus casei TISTR 1500 was isolated from soil of a dairy wastewater treatment plant and selected as the most active azo dye degrader of 19 isolates. Growing cells and freely suspended cells of this strain completely degraded methyl orange, thereby decolorizing the medium. The strain stoichiometrically converted methyl orange to N,N-dimethyl-p-phenylenediamine and 4-aminobenzenesulfonic acid, which were identified by HPLC, GC, and GC-MS analyses. The enzyme activity responsible for the cleavage of the azo bond of methyl orange was localized to the cytoplasm of cells grown on modified MRS medium containing methyl orange. The effect of sugars, oligosaccharides, organic acids, metal ions, pHs, oxygen and temperatures on methyl orange decolorization by freely suspended cells was investigated. The optimal conditions for the decolorization of methyl orange by the Lactobacillus casei TISTR 1500 are incubation at 35 degrees C and pH 6 with sucrose provided as the energy source.     

Optimisation of process parameters for bioreduction of azo dyes using Bacillus firmus under batch anaerobic condition

A new dye decolourising bacterial strain was isolated from textile wastewater and identified as Bacillus firmus. The study indicated that the bacterium could efficiently decolourise different azo dyes under static culture conditions. Characterisation of the efficiency of azo dye reduction by this isolate using both spectral and HPLC analysis was found to be a function of process parameters which include dye concentration, culture broth pH, incubation temperature, aeration as well as nitrogen source. For decolourisation, the optimal pH and temperature were 7–8 and 20–35°C respectively, while remarkable dye degradation was obtained within 18 h for dye concentrations below 100 mg L^?1. With the addition of yeast extract and under optimal conditions, dye reduction was enhanced and complete colour removal was achieved within 12 h. Colour removal was shown to be due to biodegradation rather than adsorption of dyes on bacterial cells. This study confirms the ability of the new dye‐degrading strain, Bacillus firmus, to decolourise and degrade different azo dyes and highlights its high biotechnology potential for the eco‐friendly treatment of textile wastewater when optimal conditions are applied.