Decolorization of Some Reactive Textile Dyes by White Rot Fungi Isolated in Pakistan

Summary Four white-rot fungi isolated in Pakistan were used for decolorization of widely used reactive textile dyestuffs. Phanerochaete chrysosporium, Coriolus versicolor, Ganoderma lucidum and Pleurotus ostreatus were grown in defined nutrient media for decolorization of Drimarene Orange K-GL, Remazol Brilliant Yellow 3GL, Procion BluePX-5R and Cibacron Blue P-3RGR for 10 days in shake flasks. Samples were removed every day, centrifuged and the absorbances of the supernatants were read to determine percentage decolorization. It was observed that P. chrysosporium and C. versicolor could effectively decolorize Remazol Brilliant Yellow 3GL, Procion BluePX-5R and Cibacron Blue P-3RGR. Drimarene Orange K-GL was completely decolorized (0.2 g/l after 8 days) only by P.chrysosporium, followed by P. ostreatus (0.17 g/l after 10 days). P. ostreatus also showed good decolorization efficiencies (0.19–0.2 g/l) on all dyes except Remazol Brilliant Yellow (0.07 g/l after 10 days). G. lucidum did not decolorize any of the dyestuffs to an appreciable extent except Remazol Brilliant Yellow (0.2 g/l after 8 days).     

Decolorization of synthetic dyes and production of manganese-dependent peroxidase by new fungal isolates

Two yeasts, Debaryomyces polymorphus, Candida tropicalis, and two filamentous fungi, Umbelopsis isabellina, Penicillium geastrivorus, could completely decolorize 100 mg Reactive Black 5 (RB 5) l^–1 within 16–48 h. Manganese-dependent peroxidase (MnP) activities between 60 and 424 U l^–1 were detected in culture supernatants of three of these organisms indicating the color removal by enzymatic biodegradation but with P. geastrivorus there was no ligninolytic enzyme activity in its culture and the decolorization was mainly due to biosorption to mycelium. Extensive decolorization by D. polymorphus (69–94%) and C. tropicalis (30–97%) was obtained with five other azo dyes and one anthraquinone dye. Except for Reactive Brilliant Blue KNR and Reactive Yellow M-3R, the four azo dyes, Reactive Red M-3BE, Procion Scharlach H-E3G, Procion Marine H-EXL and Reactive Brilliant Red K-2BP, induced D. polymorphus to produce MnP (105–587 U l^–1). However, MnP activities of 198–329 U l^–1 were only detected in the culture of C. tropicalis containing Reactive Red M-3BE and Reactive Brilliant Red K-2BP, respectively.     

Decolorization of Textile Reactive Dyes by Bacterial Monoculture and Consortium Screened from Textile Dyeing Effluent

Dyeing effluents have become a vital source of water pollution. Due to the xenobiotic properties and toxicity to all life forms including humans, removal of undesirable color and associated toxicity is crucial. In this study, five dye decolorizing bacteria were isolated from dyeing effluent using selective enrichment culture in Bushnell-Haas (BH) medium amended with co-substrate (i.e. glucose, yeast extract) and 100?mg?L^?1 of each commercially available reactive dyes viz. Novacron Orange FN-R, Novacron Brilliant Blue FN-R, Novacron Super Black G, Bezema Yellow S8-G and Bezema Red S2-B. The isolated bacteria were identified and assigned as Neisseria sp., Vibrio sp., Bacillus sp., Bacillus sp. and Aeromonas sp. based on their phenotypic (cultural, morphological, physiological and biochemical characteristic) observation. The dye decolorization efficiency was estimated spectrophotometrically up to 6?days of static incubation at 37?°C and observed that all of the isolates were unable to induce decolorization in absence of co-substrate. In case of monoculture, decolorization percentage varies from no visible decolorization (Bezema Red S2-B by Ek-5) to highest 90% decolorization (Novacron Brilliant Blue FN-R by Ek-13) whereas the decolorization percentage of bacterial consortium varies from 65% (Bezema Yellow S8-G) to 90% (Novacron Brilliant Blue FN-R and Novacron Super Black G). The study outlines the co-substrates mediated decolorization process where bacterial consortium proved as efficient dye decolorizer than that of the monocultures. This finding confers possibility of using novel microbial consortium for biological treatment of disreputable dyeing effluents.     

Studies on phytoremediation potentiality of Typhonium flagelliforme for the degradation of Brilliant Blue R

In vitro culture plants of Typhonium flagelliforme were found to decolorize a variety of dyes, including Malachite Green, Red HE 8B, Methyl Orange, Reactive Red 2, Direct Red 5B (DR5B), Red HE 7B, Golden Yellow HER, Patent Blue, and Brilliant Blue R (BBR), to varying extents within 4 days. The enzymatic analysis of plant roots of aseptically raised plantlets performed before and after degradation of the dye BBR by these plantlets showed a significant induction in the activities of peroxidase, laccase, tyrosinase, and 2,6-dichlorophenol-indophenol reductase, which indicated the involvement of these enzymes in the metabolism of the dye. Comparative study of the enzyme status of the plants Typhonium flagelliforme and Blumea malcolmii during the degradation of DR5B and BBR showed marked variations in the enzyme profile with respect to the use of different sources of the enzyme. Phytoremediation of BBR using Typhonium flagelliforme was confirmed with high performance liquid chromatography and Fourier transform infrared spectroscopy analysis performed before and after the degradation of the dye. One of the products of the metabolism of the dye was identified as 4-(4-ethylimino-cyclohexa-2,5-dienylidinemethyl)-phenylamine with the aid of gas chromatography–mass spectroscopy (GC–MS) analysis. Significant decrease in the American Dye Manufacturer’s Institute, biological oxygen demand, and chemical oxygen demand values of synthetic mixture of textile dyes and industrial effluent confirmed the decolorization and detoxification. Phytotoxicity studies also revealed the nontoxic nature of the metabolites of BBR.     

Increasing manganese peroxidase production and biodecolorization of triphenylmethane dyes by novel fungal consortium

A fungal consortium-SR consisting of Trametes sp. SQ01 and Chaetomium sp. R01 was developed for decolorizing three kinds of triphenylmethane dyes, which were decolorized by individual fungi with low efficiencies. The fungal consortium-SR produced 1.3 U ml(-1) of manganese peroxidase, 5.5 times higher than that produced by the monoculture of Trametes sp. SQ01, and decolorized Crystal Violet, Coomassie Brilliant Blue G250 (CBB G250) and Cresol Red. The fungal consortium-SR had a decolorization rate of 63-96%, much higher than that of the monoculture of strain SQ01 (38-72%). In consortium-SR, the higher efficiencies of decolorization of Crystal Violet and CBB G250 were obtained when they added to the culture after 4d of mixed cultivation rather than at the beginning of cultivation. Cresol Red was the exception. It is suggested that the consortium-SR has great potential for decolorizing triphenylmethane dyes.     

Dye-ligand centrifugal affinity chromatography.

A fast method for the screening of a large number of immobilized dyes for the purification or binding of proteins called dye-ligand centrifugal affinity chromatography, is described. The ease and speed of this method is demonstrated by screening 65 immobilized dyes for the binding of purified goat IgG. Two immobilized dyes (Drimarene Blue K-R and Drimarene Rubine R/K-5BL) with a high affinity for goat IgG were found to bind specifically the Fc-fragment of the IgG.     

Decolorization Screening of Synthetic Dyes by Anaerobic Methanogenic Sludge Using a Batch Decolorization Assay

The nonspecific ability of anaerobic sludge bacteria obtained from cattle dung slurry was investigated for 17 different dyes in a batch assay system using sealed serum vials. Experiments using Reactive Violet 5 (RV 5) showed that sludge bacteria could effectively decolorize solutions having dye concentrations up to 1000 mg l⁻¹ with a decolorization efficiency of above 75% during 48 h of incubation. Headspace gas composition of anaerobic batch systems for varying dye concentration revealed that lower concentrations of RV 5 (upto 500 mg l⁻¹) were found to be stimulatory to the methanogenic activity of sludge bacteria. However at higher dye concentrations, the headspace gas composition was found to be similar to batch assay controls without dye, indicating that dye at higher concentrations was inhibitory to methanogenic bacteria of sludge. The optimum inoculum and incubation temperature for maximum decolorization of RV 5 was found to be 9.0 g l⁻¹(in terms of total solids) and 37°C, respectively. Of sixteen other dyes tested, nine (Reactive Black 5, Reactive Blue 31, Reactive Blue 28, Reactive Red HE8B, Reactive Yellow, Reactive Golden Yellow, Mordant Orange, Novatic Olive R S/D & Navilan Yellow GL) were decolorized with more than 88% efficiency; three (Orange II, Navy Blue HER & Novatic Blue BC S/D) were decolorized with about 50–65% efficiency, whereas other three dyes (Procion Orange H2R, Procion Brilliant Blue HGR & Novatic Blue BC S/D) were decolorized with less than 40% efficiency. Though Ranocid Fast Blue was decolorized with about 92.5% efficiency, this was merely due to sorption, whereas the other dyes were decolorized due to biotransformation.     

Decolorization of industrial dyes by a Brazilian strain of Pleurotus pulmonarius producing laccase as the sole phenol-oxidizing enzyme

The ability of a Brazilian strain ofPleurotus pulmonarius to decolorize structurally different synthetic dyes (including azo, triphenylmethane, heterocyclic and polymeric dyes) was investigated in solid and submerged cultures. Both were able to decolorize completely or partially 8 of 10 dyes (Amido Black, Congo Red, Trypan Blue, Methyl Green, Remazol Brilliant Blue R, Methyl Violet, Ethyl Violet, Brilliant Cresyl Blue). No decolorization of Methylene Blue and Poly R 478 was observed. Of the four phenol-oxidizing enzymes tested in culture filtrates (lignin peroxidase, manganese peroxidase, aryl alcohol oxidase, laccase),P. pulmonarius produced only laccase. Both laccase activity and dye decolorization were related to glucose and ammonium starvation or to induction by ferulic acid. The decolorizationin vivo was tested using three dyes — Remazol Brilliant Blue R, Trypan Blue and Methyl Green. All of them were completely decolorized by crude extracellular extracts. Decolorization and laccase activity were equally affected by pH and temperature. Laccase can thus be considered to be the major enzyme involved in the ability ofP. pulmonarius to decolorize industrial dyes.     

Evaluation of synthetic dye decolorization capacity in Ischnoderma resinosum

The little studied white rot fungus Ischnoderma resinosum was tested for its ability to decolorize seven different synthetic dyes. The strain efficiently decolorized Orange G, Amaranth, Remazol Brilliant Blue R, Cu-phthalocyanin and Poly R-478 on agar plates and in liquid culture at a relatively high concentration of 2–4 and 0.5–1 g l⁻¹, respectively. Malachite Green and Crystal Violet were decolorized to a lower extent up to the concentration of 0.1 g l⁻¹. Decolorization capacity of I. resinosum was higher than that in Phanerochaete chrysosporium, Pleurotus ostreatus or Trametes versicolor. In contrast with these thoroughly examined fungi, I. resinosum was able to degrade a wide spectrum of chemically and structurally different synthetic dyes. I. resinosum also efficiently decolorized dye mixtures. In liquid culture, Orange G and Remazol Brilliant Blue R were decolorized most rapidly; the process was not affected by different nitrogen content in the media. Shaken cultivation strongly inhibited the decolorization of Orange G.     

Evaluation of azo dyes degradation potential of fungal strains and their role in wastewater treatment

In the present investigation, two fungal strains were exploited to evaluate their degradation capability on Synozol Red, Yellow, and Navy-Blue dyes which gave the utmost decolorization such as 40%, 70%, 90% by Aspergillus niger, and 36%, 73%, 87% by Trichoderma viride, respectively for 60 days. The Gas Chromatography-Mass Spectrometry (GC–MS) analysis of the decolorized dyes suggested that various compounds such as Caprolactam, Furazan-3-carboxamide, oxime, 4-amino-N, N-dimethyl, 6H-Pyrazolo[1,2-a] [1,2,4,5]tetrazine, Hexahydro-2,3-dimethyl, Benzene, 1-propenyl, Dihydroxymaleic acid, Arsenous acid, tris(trimethylsilyl) ester were produced by the fungi which helped in the removal of dyes from the wastewater. The laccase activity of the degraded dyes was proof that both of the strains positively produced the enzyme that helped in the biodegradation of carcinogenic dyes into less harmful products. The A. niger extracted laccase relative activity was 262%, 265%, and 145.7% for Synozol Yellow, Synozol Red, and Navy Blue, respectively. Similarly, laccase, obtained from T. viride, showed relative activity of 187.5% against Synozol Yellow, 215% against Synozol Red, and 202% against Navy Blue. Furthermore, the supernatant extracted from fungi-decolorized wastewater was used to check phytotoxicity on Vigna radiata, which gave excellent results. Both fungal strains, on the basis of their dye degradation potential, can be used to ameliorate wastewater contaminated with azo dyes.     

Synthetic dye decolorization capacity of white rot fungus Dichomitus squalens

The ability to decolorize eight chemically different synthetic dyes (Orange G, Amaranth, Orange I, Remazol Brilliant Blue R (RBBR), Cu-phthalocyanin, Poly R-478, Malachite Green and Crystal Violet) by the white rot fungus Dichomitus squalens was evaluated on agar plates. The fungus showed high decolorization capacity and was able to decolorize all dyes tested, but not to the same extent. Some of the dyes did not limit the decolorization capacity of the strain tested even at a concentration of 2g/l. The presence of the dyes in solid media reduced the mycelial growth rate of D. squalens; a positive correlation was found between the growth rate and the decolorization ability. Decolorization of Orange G and RBBR was studied also in liquid culture, where both dyes caused an enhancement of ligninolytic enzyme and overall hydrogen peroxide production and a decrease of biomass production. RBBR was removed to a higher extent than Orange G.     

Decolorization of textile dyes by whole cultures of Ischnoderma resinosum and by purified laccase and Mn-peroxidase

Ischnoderma resinosum produced extracellular ligninolytic enzymes laccase and MnP. The activity of laccase achieved the maximum on day 10 (29.4 U L⁻¹), the MnP on day 14 (34.5 U L⁻¹). Laccase and Mn-peroxidase were purified from the culture liquid using gel permeation and ion-exchange chromatographies. Purified Mn-peroxidase performed decolorization of all textile dyes tested (Reactive Black 5, Reactive Blue 19, Reactive Red 22 and Reactive Yellow 15). Laccase was inactive with Reactive Black 5 and Reactive Red 22, while all dyes were decolorized after addition of the redox mediators violuric acid (VA) and hydroxybenzotriazole (HBT). The culture liquid from I. resinosum cultures was also able to decolorize all dyes as well as the synthetic dyebaths in the presence of VA and HBT. The highest decolorization rates were detected in acidic pH (3–4).     

Decoloration of textile dyes by Trametes versicolor and its effect on dye toxicity

Amaranth, Tropaeolin O, Reactive Blue 15, Congo Red, and Reactive Black 5 were completely decolorized with no dye sorption by Trametes versicolor. Cibacron Brilliant Red 3G-P, Cibacron Brilliant Yellow 3B-A, and Remazol Brilliant Blue R were partially decolorized with some dye sorbed to the biomass. The Microtox assay before decoloration showed that Amaranth and Tropaeolin O were not toxic [the percent concentration to decrease 20% of the luminescence of Vibrio fischeri (EC₂₀) was greater than 100%]; Cibacron Brilliant Yellow 3B-A, Reactive Blue 15 and Cibacron Brilliant Red 3G-P were moderately non-toxic (100% > EC₂₀ > 75%); Remazol Brilliant Blue R was toxic (75% > EC₂₀ > 50%); and Congo Red and Reactive Black 5 were moderately toxic (50% > EC₂₀ > 25%). After decoloration the toxicity of the solutions containing Amaranth, Tropaeolin O and Reactive Black 5 was unchanged; Reactive Blue 15, Remazol Brilliant Blue R and Cibacron Brilliant Red 3G-P decreased to non-toxic levels; and Cibacron Brilliant Yellow 3B-A and Congo Red became very toxic (EC₂₀ < 25%).     

Comparison of static and shake culture in the decolorization of textile dyes and dye effluents byPhanerochœte chrysosporium

Decolorization of several dyes (Red HE-8B, Malachite Green, Navy Blue HE-2R, Magenta, Crystal Violet) and an industrial effluent with growing cells ofPhanerochœte chrysosporium in shake and static culture was demonstrated. All the dyes and the industrial effluent were decolorized to some extent with varying percentages of decolorization (20–100%). The rate of decolorization was very rapid with Red HE-8B, an industrial dye. Decolorization rates for all the dyes in static condition were found to be less than the shake culture and also dependent on biomass concentration.     

Decolorization of triphenylmethane and azo dyes by Citrobacter sp.

A Citrobacter sp., isolated from soil at an effluent treatment plant of a textile and dyeing industry, decolorized several recalcitrant dyes except Bromophenol Blue. More than 90% of Crystal Violet and Methyl Red at 100 M were reduced within 1 h. Gentian Violet, Malachite Green and Brilliant Green lost over 80% of their colors in the same condition, but the percentage decolorization of Basic Fuchsin and Congo Red were less than the others, 66 and 26%, respectively. Decolorization of Congo Red was mainly due to adsorption to cells. Color removal was optimal at pH 7–9 and 35–40 °C. Decolorization of dyes was also observed with extracellular culture filtrate, indicating the color removal by enzymatic biodegradation.     

Decolorization of reactive dyes by the white rot fungus Trametes versicolor in sequencing batch reactors

The white rot fungus Trametes versicolor was shown to be capable of decolorizing three reactive dyes in a sequencing batch process, using glucose as the carbon and energy source over an extended period without supplementation of new mycelium. Decolorization activity was related to the expression of extracellular peroxidases and could be continuously reactivated by sheering the suspended pellets. Pure culture experiments were carried out simultaneously in agitated Erlenmeyer flasks and in completely stirred tank reactors with two azo dyes, C.I. Reactive Black 5 and C.I. Reactive Red 198 as well as the anthraquinone dye C.I. Reactive Blue 19 (Brilliant Blue R). Results show high and stable degrees of decolorization of 91%-99% in both systems, which could be repeated without decrease in activity over time. Under nonsterile conditions only five cycles of decolorization could be achieved. An increasing bacterial population suppressed fungal growth and the formation of peroxidases. Copyright John Wiley & Sons, Inc.     

Contributions of functional groups and extracellular polymeric substances on the biosorption of dyes by aerobic granules

The contributions of loosely bound extracellular polymeric substances (LB-EPS), tightly bound EPS (TB-EPS), residual sludge (the sludge left after EPS extraction) and functional groups such as amine, carboxyl, phosphate and lipid on aerobic granules on biosorption of four different dyes (Reactive Brilliant Blue KN-R (KN-R), Congo Red (CR), Reactive Brilliant Red K-2G (RBR) and Malachite Green (MG)) were investigated. EPS may be responsible for biosorption of cationic dyes. However, residual sludge always made greater contribution than that of EPS. The biosorption mechanisms were dependent on the functional groups on aerobic granules and dyes’ chemical structures. The lipid and phosphate groups might be the main binding sites for KN-R biosorption. Amine, carboxyl, phosphate and lipid were all responsible for the binding of CR. The lipid fractions played an important role for RBR biosorption. For MG, the phosphate groups gave the largest contribution.     

Decolorization of synthetic textile dyes by lignin peroxidase of Phanerochaete chrysosporium

Neem hull waste (containing a high amount of lignin and other phenolic compounds) was used for lignin peroxidase production byPhanerochaete chrysosporum under solid-state fermentation conditions. Maximum decolorization achieved by partially purified lignin peroxidase was 80% for Porocion Brilliant Blue HGR, 83 for Ranocid Fast Blue, 70 for Acid Red 119 and 61 for Navidol Fast Black MSRL. The effects of different concentrations of veratryl alcohol, hydrogen peroxide, enzyme and dye on the efficiency of decolorization have been investigated. Maximum decolorization efficiency was observed at 0.2 and 0.4 mmol/L hydrogen peroxide, 2.5 mmol/L veratryl alcohol and pH 5.0 after a 1-h reaction, using 50 ppm of dyes and 9.96 mkat/L of enzyme.     

一株高效广谱染料降解细菌的分离鉴定及脱色特性研究

通过梯度驯化,从印染废水长期污染土壤中分离筛选出能以4种不同结构类型的染料(刚果红、美蓝、孔雀绿和活性艳蓝KN-R)为唯一碳源的菌株XSMR,根据其形态学特征和生理生化鉴定及16S rDNA序列分析,初步鉴定为无色杆菌属(Achromobacter sp.)的菌株。菌株XSMR对4种染料均具有强的脱色降解能力,且对染料脱色的同时,自身能够生长繁殖,培养24h菌体干重超过不加染料的对照。在振荡培养条件下对该菌株的脱色反应条件进行研究,结果表明,当刚果红、美蓝、孔雀绿及活性艳蓝KN-R的初始浓度分别小于200mg/L、200mg/L、150mg/L及150mg/L时,在pH7.5、温度35℃、接种量4%(V/V)条件下,接种菌株XSMR脱色14h对4种染料的脱色率均可达到98%以上。通过对降解产物的紫外-可见光谱分析,进一步证明了菌株XSMR能彻底降解染料。菌株XSMR对染料脱色的机理包括生物降解和菌株吸附两方面。     

Decolorization of anthraquinone dye by Shewanella decolorationis S12

A new species of genus Shewanella, Shewanella decolorationis S12, from activated sludge of a textile-printing wastewater treatment plant, can decolorize Reactive Brilliant Blue K-GR, one kind of anthraquinone dye, with flocculation first. Although S. decolorationis displayed good growth in an aerobic condition, color removal was the best in an anaerobic condition. For color removal, the most suitable pH values and temperatures were pH 6.0–8.0 and 30–37°C under anaerobic culture. More than 99% of Reactive Brilliant Blue K-GR was removed in color within 15 h at a dye concentration of 50 mg/l. Lactate was the suitable carbon source for the dye decolorization. A metal compound, HgCl₂, had the inhibitory effect on decolorization of Reactive Brilliant Blue K-GR, but a nearly complete decolorization also could be observed at a HgCl₂ concentration of 10 mg/l. The enzyme activities, which mediate the tested dye decolorization, were not significantly affected by preadaptation of the bacterium to the dye.