Decolorization of azo dyes and simulated dye bath wastewater using acclimatized microbial consortium--biostimulation and halo tolerance

Anaerobic acclimatization of activated sludge from a textile effluent treatment plant to high concentration of RB5 could effectively decolorize RB5 dye solution. The strains viz. Pseudomonas aeruginosa and Bacillus circulans and other unidentified laboratory isolates (NAD1 and NAD6) were predominantly present in the microbial consortium. The conditions for efficient decolorization, biostimulation to increase effectiveness of microbial consortium, its tolerance to high salt concentration and non-specific ability towards decolorization of eight azo dyes, are reported. The optimum inoculums concentration for maximum decolorization were found to be 1-5 ml of 1800+/-50 mg l(-1) MLSS and 37 degrees C, respectively. The decolorization efficiency was 70-90% during 48 h. The biomass showed efficient decolorization even in the presence of 10% NaCl, as tested with RB5. In the presence of flavin adenine dinucleotide (FAD) more than 99% decolorization occurred in 8h. The decolorization of RB5 was traced to extracellular enzymes. The effectiveness of acclimatized biomass under optimized conditions towards decolorization of two types of synthetic dye bath wastewaters that were prepared using chosen azo dyes is reported.     

Azo dye decolorization with a mutant Escherichia coli strain

A recombinant Escherichia coli strain (E. coli NO3) containing genomic DNA fragments from azo-reducing wild-type Pseudomonas luteola strain decolorized a reactive azo dye (C.I. Reactive Red 22) at approx. 17 mg dye h^–1 g cell. The ability to decolorize the azo dye probably did not originate from the plasmid DNA. Acclimation in azo-dye-containing media gave a nearly 10% increase in the decolorization rate of E. coli NO3. Growth with 1.25 g glucose l^–1 completely stopped the decolorization activity. When the decolorization metabolites from E. coli NO3 were analyzed by HPLC and MS, the results suggested that decolorization of the azo dye may be due to cleavage of the azo bond.     

Fe(III)-enhanced Azo Reduction by <Emphasis Type="Italic">Shewanella decolorationis</Emphasis> S12

Shewanella decolorationis S12 is capable of high rates of azo dye decolorization and dissimilatory Fe(III) reduction. Under anaerobic conditions, when Fe(III) and azo dye were copresent in S12 cultures, dissimilatory Fe(III) reduction and azo dye biodecolorization occurred simultaneously. Furthermore, the dye decolorization was enhanced by the presence of Fe(III). When 1 mM Fe(III) was added, the methyl red decolorizing efficiency was 72.1% after cultivation for 3 h, whereas the decolorizing efficiency was only 60.5% in Fe(III)-free medium. The decolorizing efficiencies increased as the concentration of Fe(III) was increased from 0 to 6 mM. Enzyme activities, which mediate the dye decolorization and Fe(III) reduction, were not affected by preadaption of cells to Fe(III) and azo dye nor by the addition of chloramphenicol. Both the Fe(III) reductase and the azo reductase were membrane associated. The respiratory electron transport chain inhibitors metyrapone, dicumarol, and stigmatellin showed significantly different effects on Fe(III) reduction than on azo dye decolorization.     

Semicontinuous decolorization of azo dyes by rotating disc contactor immobilized with<Emphasis Type="Italic">Aspergillus sojae</Emphasis> B-10

Aspergillus sojae B-10 was immobilized and used to treat model dye compounds. The model wastewater, containing 10 ppm of azo dyes such as Amaranth, Sudan III, and Congo Red, was treated with cells attached to a rotating disc contactor (RDC). Amaranth was decolorized more easily than were Sudan III and Congo Red. Decolorization of Amaranth began within a day, and the dye was completely decolorized within 5 days of incubation. Both Sudan III and Congo Red were almost completely decolorized after 5 days of incubation. Semicontinuous decolorization of azo by reusing attached mycelia resulted in almost complete decolorization in 20 days. This experiment indicated that decolorization was successfully conducted by removing azo dyes withAspergillus sojae B-10.     

Degradation of Remazol Red dye by <Emphasis Type="Italic">Galactomyces geotrichum</Emphasis> MTCC 1360 leading to increased iron uptake in <Emphasis Type="Italic">Sorghum vulgare</Emphasis> and <Emphasis Type="Italic">Phaseolus mungo</Emphasis> from soil

Removal of azo dyes from the effluent generated by textile industries is rather difficult. Azo dyes represent a major class of synthetic colorants that are both mutagenic and carcinogenic. Galactomyces geotrichum MTCC 1360, a yeast species, showed more than 96% decolorization of the azo dye Remazol Red (50 mg/L) within 36 h at 30°C and pH 11.0 under static condition with a significant reduction in the chemical oxygen demand (62%) and total organic carbon (41%). Peptone (5.0 g/L), rice husk (10 g/L extract), and ammonium chloride (5.0 g/L) were found to be more significant among the carbon and nitrogen sources used. The presence of tyrosinase, NADH-DCIP reductase, riboflavin reductase and induction in azo reductase and laccase activity during decolorization indicated their role in degradation. High performance thin layer chromatography analysis revealed the degradation of Remazol Red into different metabolites. Fourier transform infrared spectroscopy and high performance liquid chromatography analysis of samples before and after decolorization confirmed the biotransformation of dye. Atomic absorption spectroscopy analysis revealed a less toxic effect of the metabolites on iron uptake by Sorghum vulgare and Phaseolus mungo than Remazol Red dye. Remazol Red showed an inhibitory effect on iron uptake by chelation and an immobilization of iron, whereas its metabolites showed no chelation as well as immobilization of iron. Phytotoxicity study indicated the conversion of complex dye molecules into simpler oxidizable products which had a less toxic nature.     

A batch decolorization and kinetic study of Reactive Black 5 by a bacterial strain Enterobacter sp. GY-1

An Enterobacter strain (GY-1) with high activity of decolorization of Reactive Black 5 (RB 5) was isolated from textile wastewater treating sludge. The kinetic characteristics of dye decolorization by the strain GY-1 were determined quantitatively using the diazo dye, RB 5. Effects of different operation parameters (inoculum size, pH, temperature and salinity) and various electron donors on decolorization of the azo dye by GY-1 were systematically investigated to reveal the primary factors that determine the performance of the azo dye decolorization. The decolorization of RB 5 was attributed to extracellular enzymes. A kinetic model was established giving the dependence of decolorization rate on cell mass concentration (first order). Decolorization rate increased with increasing temperature from 20 to 35 °C, which can be predicted by Arrhenius equation with the activation energy (E_a) of 8.50 kcal mol⁻¹ and the frequency factor (A₀) of 6.28 × 10⁷ mg l g MLSS⁻¹ h⁻¹. Michaelis-Menten kinetics and Eadie-Hofstee plot were used to determine V_max, 1.05 mg l⁻¹ h⁻¹ and K_m, 24.06 mg l⁻¹.     

Decolorization of Fast red by metabolizing cells of <Emphasis Type="Italic">Oenococcus oeni</Emphasis> ML34

Three different azo dyes such as Fast red, metanil yellow and Fast orange were examined for their decolorization by O. oeni ML34. Fast red (FR) was decolorized by 68%, whereas the other dyes were removed by only about 30%. The effects of glucose addition, substrate (dye) concentration and environmental factors (temperature, pH) on decolorization were investigated by two-level factorial design. The statistical analyses revealed that glucose specifically increases the extent of FR decolorization. A glucose level of 5 g/l was the optimum concentration for removal of, FR reaching a decolorization percentage of up to 93%.     

Biodecolorization of textile azo dyes by isolated yeast from activated sludge: Issatchenkia orientalis JKS6

An ascomycetous yeast strain isolated from activated sludge could decolorize Reactive Black 5 azo dye at 200 mg l^?1 up to 90 % within 12–18 h under agitated condition. Yeast decolorization ability was investigated at different RB5 concentrations and, at higher dye concentration, 500 mg l^?1, the decolorization was found to be 98 % after 36 h incubation time. Extensive decolorization (95–99 %) was obtained in presence of five other azo dyes, Reactive Orange 16, Reactive Red 198, Direct Blue 71, Direct Yellow 12, and Direct Black 22, by isolated yeast. HPLC analysis, UV–vis spectra and colorless biomass obtained after complete decolorization showed that the decolorization occured through a biodegradation mechanism. Decolorization was occurred during the exponential growth phase which is associated to primary metabolism. Laccase production by the yeast cells was not detected. The isolated yeast was characterized according to phenotypical and molecular procedures and was closely related (99 % identity) to Issatchenkia orientalis.     

Differential Expression of Antioxidant Enzymes During Degradation of Azo Dye Reactive black 8 in Hairy roots of Physalis minima L.

The enzymes involved in the protection of plant metabolism in presence of azo dye was characterized by studying activities of the role of antioxidant enzymes in the hairy roots (HRs) of Physalis minima L. during degradation of an azo dye, Reactive Black 8 (RB8). When the HRs were exposed to RB8 (30 mg L^?1), a nine fold increase in SOD activity was observed after 24 h, while 22 and 50 fold increase in activity was observed for POX and APX respectively after 72 h, whereas there was no significant change in activity of CAT. The activation of different antioxidant enzymes at different time intervals under dye stress suggests the synchronized functioning of antioxidant machinery to protect the HRs from oxidative damage. FTIR analysis confirmed the degradation of dye and the non-toxic nature of metabolites formed after dye degradation was confirmed by phytotoxicity study.     

绒毛栓孔菌菌丝体在无营养条件下对偶氮染料刚果红的脱色作用

【目的】在无营养条件下,利用白腐真菌绒毛栓孔菌(Trametes pubescens)菌丝体对染料进行脱色可减少试验成本,提高染料处理的实用性。【方法】将该菌株液体培养的菌丝体在无营养条件下对染料进行脱色,并对其中脱色效果较好的偶氮染料刚果红的脱色过程进行分析。在此过程中,测定了该菌株分泌的胞外胞内酶活力,优化影响因子如初始pH值、温度、染料浓度和盐度,同时利用气相色谱-质谱联用技术分析无营养条件下偶氮染料刚果红的降解产物。植物毒性试验测定刚果红经绒毛栓孔菌菌丝体脱色前后的毒性变化。【结果】菌丝体对偶氮染料刚果红有较好的脱色效果,在初始pH值为2.0,温度为30°C,染料浓度为80 mg/L,盐度为2.5%(质量体积比)时,150 r/min转速下培养7 d后脱色率可达80.52%。在此过程中,菌丝体可被连续使用2次,且其所分泌的酶系可降解染料。此外,通过气相色谱-质谱联用分析得到刚果红的降解产物为萘胺、联苯胺和叠氮萘。植物毒性试验显示在无营养条件下的绒毛栓孔菌菌丝体对染料有明显的脱毒作用。【结论】研究发现绒毛栓孔菌菌丝体在无营养条件下的偶氮染料废水处理中具有广阔的应用前景。     

Decolorization of azo and triphenylmethane dyes by Pycnoporus sanguineus producing laccase as the sole phenoloxidase

Partial decolorization of two azo dyes (orange G and amaranth) and complete decolorization of two triphenylmethane dyes (bromophenol blue and malachite green) was achieved by cultures in submerged liquid culture producing laccase as the sole phenoloxidase. Enzyme production could be correlated with dye decolorization, with sorption of dye to mycelia accounting for less than 3% of dye removal.     

Decolorization of chemically different dyes by white-rot fungi in submerged cultures

Forty-two white-rot fungi in submerged cultures were tested to determine their dye decolorization capacity and the optimal conditions for the decolorization process. Trametes pubescens Cui 7571 was found to be the most effective strain in terms of decolorization performance on the azo dye Congo Red, and it exhibited excellent reusability as well as persistence in sequential decolorization experiments. Optimization of the decoloration process was also conducted to evaluate the effects of a number of chemical compounds, metal salts, inducers, and mediators on the dye decolorization rate. On the seventh day, a highest dye removal of 98.83 % was observed with addition of copper at 2.5 mmol L^?1, Tween 80 at 1.0 % (v/v), and ferulic acid at 0.50 μmol L^?1, respectively. The adsorption of mycelia to dyes was not a significant contributor to dye removal, and decolorization by the functional fungus T. pubescens depended on biodegradation by enzymes, as evidenced by the results of the moist heat sterilization treatment (121°C for 20 min), induction of extracellular enzymes, and scanning electron microscopy. Four dye degradation metabolites, i.e., naphthalene amine, biphenyl amine, biphenyl ,and naphthalene diazonium, were identified by Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry. The phytotoxicity tests indicated that degraded metabolites had almost a negligible effect on the plant seeds as compared to that of dye, which is indicative of the less toxic nature of the metabolites. Our results suggest that white-rot fungus T. pubescens could be developed into a novel azo dye bioremediation strategy.     

Decolorization of azo dyes by <Emphasis Type="Italic">Shewanella oneidensis</Emphasis> MR-1 in the presence of humic acids

The effects of humic acid (HA) on azo dye decolorization by Shewanella oneidensis MR-1 were studied. It was found that HA species isolated from different sources could all accelerate the decolorization of Acid Red 27 (AR27). Anoxic and anaerobic conditions were required for the enhancement of azo dye decolorization by HA. In the presence of 50 mg DOC L⁻¹ Aldrich HA, 15–29% increases in decolorization efficiencies of azo dyes with different structures were achieved in 11 h. The enhancing effects increased with the increase of HA concentrations ranging from 25 to 150 mg DOC L⁻¹, and the decolorization rates were directly proportional to the HA concentrations when they were below 100 mg DOC L⁻¹. Lactate and formate were good electron donors for AR27 decolorization in the presence of HA. Both nitrate (0.1–3.0 mM) and nitrite (0.3–1.2 mM) inhibited AR27 decolorization in the presence of HA, and negligible decolorization was observed before their removal. Soluble FeCl₃ could accelerate the decolorization process in the presence of HA, whereas insoluble hematite could not. These findings may affect the understanding of bioremediation of azo dye-polluted environments and help improve the treatment of azo dye wastewaters.     

Decolorization of sulfonated azo dyes with two photosynthetic bacterial strains and a genetically engineered Escherichia coli strain

Sulfonated azo dyes were decolorized by two wild type photosynthetic bacterial (PSB) strains (Rhodobacter sphaeroides AS1.1737 and Rhodopseudomonas palustris AS1.2352) and a recombinant strain (Escherichia coli YB). The effects of environmental factors (dissolved oxygen, pH and temperature) on decolorization were investigated. All the strains could decolorize azo dye up to 900 mg l⁻¹, and the correlations between the specific decolorization rate and dye concentration could be described by Michaelis–Menten kinetics. Repeated batch operations were performed to study the persistence and stability of bacterial decolorization. Mixed azo dyes were also decolorized by the two PSB strains. Azoreductase was overexpressed in E. coli YB; however, the two PSB strains were better decolorizers for sulfonated azo dyes.     

Bacterial Decolorization of Azo Dyes by Rhodopseudomonas palustris

Summary The ability of Rhodopseudomonas palustris AS1.2352 possessing azoreductase activity to decolorize azo dyes was investigated. It was demonstrated that anaerobic conditions were necessary for bacterial decolorization, and the optimal pH and temperature were pH 8 and 30–35 °C, respectively. Decolorization of dyes with different molecular structures was performed to compare their degradability. The strain could decolorize azo dye up to 1250 mg l⁻¹, and the correlation between the specific decolorization rate and dye concentration could be described by Michaelis–Menten kinetics. Long-term repeated operations showed that the strain was stable and efficient during five runs. Cell extracts from the strain demonstrated oxygen-insensitive azoreductase activity in vitro.     

The implication of Dichomitus squalens laccase isoenzymes in dye decolorization by immobilized fungal cultures

The study focuses on the production of ligninolytic enzymes and dye degradation capacity of Dichomitus squalens immobilized on polyurethane foam (PUF) or pine wood (PW) in a fixed bed reactor at a laboratory scale (working volume of 0.6l). Immobilization of fungal cultures on pine wood improved eminently laccase production in comparison to the liquid cultures. Immobilized D. squalens was able to decolorize an anthraquinone dye Remazol Brilliant Blue R and an azo dye Reactive Orange 16, however, only a limited decolorization of Copper(II)phthalocyanine dye was observed in both types of reactor cultures. The involvement of a laccase activity in dye decolorization was suggested. Further, two different chromatographical forms of laccases, Lc1 and Lc2, were isolated from PW cultures of D. squalens using a fast, two step FPLC method. Enzymes revealed identical molecular masses of 68 kDa (estimated by SDS-PAGE) and similar pI's, however, they differed in their catalytic properties such as pH dependence of the activity and ABTS oxidation rates. In this study, we demonstrated different dye decolorization capacities of Lc1 and Lc2 as well.     

氧气对混合菌群脱色降解偶氮染料效果的影响

【背景】偶氮染料及其中间产物具有一定的环境毒性,利用混合菌群降解偶氮染料是一种环境友好型方法,但降解过程中氧气的存在起到至关重要的作用,可以促进或抑制偶氮染料的微生物降解作用。【目的】探讨氧气对偶氮染料微生物脱色液的影响,分析氧气对混合菌群脱色降解偶氮染料效果的影响。【方法】利用混合菌群DDMY1在3种培养条件(好氧、厌氧、兼氧)下,对7种偶氮染料进行脱色降解,探讨偶氮染料脱色液对氧气的响应情况,利用紫外可见分光光度法(ultraviolet visible spectrophotometry,UV-vis)和傅里叶变换红外光谱法(Fourier transform infrared spectroscopy,FTIR)对脱色产物进行分析。【结果】在兼氧和厌氧条件下反应48 h后的染料脱色液,与氧气充分接触后,部分偶氮染料微生物脱色液发生较为明显的复色现象,如活性黑5、直接黑38;UV-vis分析结果表明,这种复色现象是由于脱色液与氧气接触之后产生新物质所致;FTIR分析结果表明,混合菌群对发生复色反应的偶氮染料仍然具有一定脱色降解效果,但是脱色尚不够完全。【结论】兼氧和厌氧条件下,氧气对部分偶氮染料微生物脱色液具有较为明显的影响,从而影响混合菌群对偶氮染料的整体脱色效果,这可为今后研究偶氮染料彻底生物降解提供理论基础。     

Decolorization of Acid Black 210 by <Emphasis Type="Italic">Vibrio harveyi</Emphasis> TEMS1, a newly isolated bioluminescent bacterium from Izmir Bay,Turkey

The present study deals with the decolorization of Acid Black 210 by a bioluminescent bacterium, Vibrio harveyi TEMS1, isolated from coastal seawater of Izmir Bay, Turkey. Maximum rate of decolorization of Acid Black 210 was observed when Luria Bertani medium was used. Decolorization of Acid Black 210 was 38.9% and 93.9% at 24 h under shaking and static conditions, respectively. The optimum dye-decolorizing activity of the culture was obtained at 100 ppm initial dye concentration and incubation temperature of 20°C. Vibrio harveyi TEMS1 was also tested for its ability to decolorize four azo dyes (Acid Black 24, Acid Blue 7, Acid Green 20, Acid Yellow 36) in addition to Acid Black 210.     

Biodegradation of Reactive Blue 59 by isolated bacterial consortium PMB11

Morphologically different, three bacterial strains, capable of decolorizing Reactive Blue 59 were isolated from dye effluent contaminated soil sample, collected from Ichalkaranji, India. The individual bacterial strains viz. Bacillus odysseyi SUK3, Morganella morganii SUK5 and Proteus sp. SUK7 decolorized Reactive Blue 59 (50 mg l(-1)) completely within 60, 30, 24 h, respectively, while the bacterial consortium PMB11 of these strains required 3 h for the complete decolorization. The decolorization was confirmed by UV-Vis spectroscopy. Further, the biodegradation of Reactive Blue 59 in to different metabolites was confirmed by High performance liquid chromatography and Fourier transform infrared spectroscopy analysis. Significant increase in the activity of aminopyrine N-demethylase (AND) in the individual as well consortium cells, obtained after decolorization showed involvement of AND in the decolorization process. Phytotoxicity studies, revealed the nontoxic nature of the degraded metabolites of Reactive Blue 59 indicating effectiveness of bacterial consortium PMB11 for the treatment of textile effluent containing Reactive Blue 59.     

Decolorization kinetics of the azo dye Reactive Red 2 under methanogenic conditions: effect of long-term culture acclimation

The biological decolorization of the textile azo dye Reactive Red 2 was investigated using a mixed, mesophilic methanogenic culture, which was developed with mixed liquor obtained from a mesophilic, municipal anaerobic digester and enriched by feeding a mixture of dextrin/peptone as well as media containing salts, trace metals and vitamins. Batch decolorization assays were conducted with the unacclimated methanogenic culture and dye decolorization kinetics were determined as a function of initial dye, biomass, and carbon source concentrations. Dye decolorization was inhibited at initial dye concentrations higher than 100 mg l^-1 and decolorization kinetics were described based on the Haldane model. The effect of long-term culture exposure to the reactive dye on decolorization kinetics, culture acclimation, as well as possible dye mineralization was tested using two reactors fed weekly for two years with an initial dye concentration of 300 mg l^-1 and a mixture of dextrin/peptone. The maximum dye decolorization rate after a 2-year acclimation at an initial dye concentration of 300 mg l^-1 was more than 10-fold higher as compared to that obtained with the unacclimated culture. Aniline and the o-aminohydroxynaphthalene derivative resulting from the reductive azo bond cleavage of the dye were detected, but further transformation(s) leading to dye mineralization were not observed. Reactive Red 2 did not serve as the carbon and energy source for the mixed culture, and dye decolorization was sustained by the continuous addition of dextrin and peptone. Thus, biological decolorization of reactive azo dyes is feasible under conditions of low redox potential created and maintained in overall methanogenic systems, but supply of a biodegradable carbon source is necessary.