Unraveling the molecular response of Brassica napus hairy roots in the active Naphthol blue-black removal: Insights from proteomic analysis.

In vitro plant cultures are able to remove and metabolise xenobiotics, making them promising tools for decontamination strategies. In this work, we evaluated Brassica napus hairy roots (HRs) to tolerate and remove high concentrations of the azo dye Naphthol Blue-Black (NBB). Experiments were performed using both growing and resting culture systems at different pHs. Reuse of HRs biomass was evaluated in successive decolourisation cycles. Proteomics was applied to understand the molecular responses likely to be involved in the tolerance and removal of NBB. The HRs tolerated up to 480 µg mL-1 NBB, and 100 % removal was achieved at 180 µg mL-1 NBB after 10 days using both culture systems. Interestingly, the HRs are robust enough to be reused, showing 55-60 % removal even after three reuse cycles. The highest dye removal rates were achieved during the first 2 days of incubation, as initial removal is mainly driven by passive processes. Active mechanisms are triggered later by regulating the expression of proteins with different biological functions, mainly those related to xenobiotic metabolism, such as hydrolytic and redox enzymes. These results suggest that B. napus HRs are a robust tool that could make a significant contribution to textile wastewater treatment.

Removal of Azo Dyes from Water Using Natural Luffa cylindrica as a Non-Conventional Adsorbent.

Reducing high concentrations of pollutants such as heavy metals, pesticides, drugs, and dyes from water is an emerging necessity. We evaluated the use of Luffa cylindrica (Lc) as a natural non-conventional adsorbent to remove azo dye mixture (ADM) from water. The capacity of Lc at three different doses (2.5, 5.0, and 10.0 g/L) was evaluated using three concentrations of azo dyes (0.125, 0.250, and 0.500 g/L). The removal percent (R%), maximum adsorption capacity (Qm), isotherm and kinetics adsorption models, and pH influence were evaluated, and Fourier-transform infrared spectroscopy and scanning electron microscopy were performed. The maximum R% was 70.8% for 10.0 g L-1Lc and 0.125 g L-1 ADM. The Qm of Lc was 161.29 mg g-1. Adsorption by Lc obeys a Langmuir isotherm and occurs through the pseudo-second-order kinetic model. Statistical analysis showed that the adsorbent dose, the azo dye concentration, and contact time significantly influenced R% and the adsorption capacity. These findings indicate that Lc could be used as a natural non-conventional adsorbent to reduce ADM in water, and it has a potential application in the pretreatment of wastewaters.

Deciphering the origin of the first steps in the degradation of azo dyes: a computational study.

Azo dyes find applications across various sectors including food, pharmaceuticals, cosmetics, printing, and textiles. The contaminating effects of dyes on aquatic environments arise from toxic effects caused by their long-term presence in the environment, buildup in sediments, particularly in aquatic species, degradation of pollutants into mutagenic or mutagenic compounds, and low aerobic biodegradability. Therefore, we theoretically propose the first steps of the degradation of azo dyes based on the interaction of hydroperoxyl radical (•OOH) with the dye. This interaction is studied by the OC and ON mechanisms in three azo dyes: azobenzene (AB), disperse orange 3 (DO3), and disperse red 1 (DR1). Rate constants calculated at several temperatures show a preference for the OC mechanism in all the dyes with lower activation energies than the ON mechanism. The optical properties were calculated and because the dye-•OOH systems are open shell, to verify the validity of the results, a study of the spin contamination of the ground [Formula: see text] and excited states [Formula: see text] was previously performed. Most of the excited states calculated are acceptable as doublet states. The absorption spectra of the dye-•OOH systems show a decrease in the intensity of the bands compared to the isolated dyes and the appearance of a new band of the type π → π* at a longer wavelength in the visible region, achieving up to 868 nm. This demonstrates that the reaction with the •OOH radical could be a good alternative for the degradation of the azo dyes.

Changing Despicable Me: Potential replacement of azo dye yellow tartrazine for pequi carotenoids employing ionic liquids as high-performance extractors.

Color is a crucial sensory attribute that guides consumer expectations. A high-performance pequi carotenoid extraction process was developed using ionic liquid-based ethanolic solutions and a factorial design strategy to search for a potential substitute for the artificial azo dye yellow tartrazine. All-trans-antheraxanthin was identified with HPLC-PAD-MSn for the first time in pequi samples. [BMIM][BF4] was the most efficient ionic liquid, and the maximization process condition was the solid-liquid ratio R(S/L) of 1:3, the co-solvent ratio R(IL/E) of 1:1 ([BMIM][BF4]: ethanol), and three cycles of extraction with 300 s each and yielded 107.90 µg carotenoids/g of dry matter. The ionic liquid-ethanolic solution recyclability was accomplished by freezing and precipitating with an average recovery of 79 %. In CIELAB parameters, pequi carotenoid extracted with [BMIM][BF4] was brighter and yellower than the artificial azo dye yellow tartrazine. A color change of 11.08 and a hue* difference of 1.26° were obtained. Furthermore, carotenoids extracted with [BMIM][BF4] showed antioxidant activity of 35.84 µmol of α-tocopherol. These findings suggest the potential of employing the pequi carotenoids to replace the artificial azo dye yellow tartrazine in foods for improved functional properties.

Harnessing Efficient ROS Generation in Carbon Dots Derived from Methyl Red for Antimicrobial Photodynamic Therapy.

The emergence of drug-resistant pathogenic microorganisms has become a public health concern, with demand for strategies to suppress their proliferation in healthcare facilities. The present study investigates the physicochemical and antimicrobial properties of carbon dots (CD-MR) derived from the methyl red azo dye. The morphological and structural analyses reveal that such carbon dots present a significant fraction of graphitic nitrogen in their structures, providing a wide emission range. Based on their low cytotoxicity against mammalian cells and tunable photoluminescence, these carbon dots are applied to bioimaging in vitro living cells. The possibility of using CD-MR to generate reactive oxygen species (ROS) is also analyzed, and a high singlet oxygen quantum efficiency is verified. Moreover, the antimicrobial activity of CD-MR is analyzed against pathogenic microorganisms Staphylococcus aureus, Candida albicans, and Cryptococcus neoformans. Kirby-Bauer susceptibility tests show that carbon dots synthesized from methyl red possess antimicrobial activity upon photoexcitation at 532 nm. The growth inhibition of C. neoformans from CD-MR photosensitization is investigated. Our results show that N-doped carbon dots synthesized from methyl red efficiently generate ROS and possess a strong antimicrobial activity against healthcare-relevant pathogens.

Dietary exposure to food azo-colours in a sample of pre-school children from Southern Brazil.

The dietary exposure of six food azo-colours was assessed in a sample of pre-schoolers from Guaratuba-Paraná, Brazil. Consumption data of 323 children aged 2 to 5 years was collected through 3-day food records. Dietary exposure, is expressed by milligrams of food colour by kilogram of body weight per day, as compared to the Acceptable Daily Intake (ADI). Three exposure scenarios were developed to account for uncertainties around consumption estimates. Intakes of Amaranth (INS 123) described in means, 50th and 95th percentiles exceeded ADI levels in the two most conservative scenarios, with the highest percentiles exceeding about four times the ADI. High intakes of Sunset Yellow FCF (INS 110) were also observed, of up to 85% of the ADI in the worst-case scenario. Findings suggest high exposure levels to azo-dyes in the survey population, with children likely exceeding the ADI for Amaranth (INS 123) and concerns for Sunset Yellow FCF (INS 110). Major food contributors were beverages (juice powders and soft drinks), dairy and sweets. Further studies on dietary exposure assessment are needed at the national level. The authors highlight the need of controlling the use of such additives through national policies that are aligned with the consumption patterns observed in the country.

Non-regulated aromatic amines in clothing purchased in Spain and Brazil: Screening-level exposure and health impact assessment.

The staggering amount of chemicals in clothes and their harmful effects on human health and the environment have attracted the attention of regulatory agencies and the scientific community worldwide. Azo dyes are synthetic dyestuffs with widespread use in textile industries, currently classified as emerging pollutants of great health concern to consumers. These compounds may release one or more aromatic amines (AAs) after reductive cleavage of their azo bounds. Twenty-two AAs have already been regulated due to their carcinogenic effects. However, since information on their potential toxicity is not currently available, several AAs have not been still regulated by the European Union. Considering this gap, the present study aimed to assess the levels of forty non-regulated AAs in 240 clothing items from Spain and Brazil. The potential impact on the health of vulnerable population groups after dermal exposure to those garments was also evaluated. In Brazil, at least one AA was detected in the clothes, while in samples obtained in Spain, only two of them showed values below the limit of detection for AAs. In 75 clothes, at least one of the measured AAs was higher than the hazardous threshold (30 mg/kg), which can mean risks to human health since these compounds are suspected to be mutagenic. Aniline, the most common AA, showed a high detection rate (82%) in clothes, with significantly higher concentrations in items commercialized in Brazil (0.35 vs. 0.17 mg/kg; p = 0.032). Moreover, o-aminobenzenesulfonic and p-phenylenediamine, suspected mutagenic, were found at relevant concentrations in several clothes, mainly made of synthetic fibers. In this study, the hazard index associated with exposure to AAs through clothing was low (0.006-0.13) for all the population groups of both countries in the medium-bound scenario. However, its value was close to 1 for Brazilian pregnant women (0.998) when the maximum concentration value was considered under an upper-bound scenario. The risk of exposure to non-regulated AAs may be underestimated since only dermal exposure was considered for risk assessment. Moreover, the co-occurrence of other carcinogenic and non-carcinogenic substances present in skin-contact clothes should mean an additional source of potential risk.

Screening of regulated aromatic amines in clothing marketed in Brazil and Spain: Assessment of human health risks.

Azo dyes used in textile products contain aromatic amines (AAs), which may be released into the environment after skin bacteria cleavage the azo bond. In Europe, 22 carcinogenic AAs are regulated. Unfortunately, no information is available in many non-European countries, including Brazil. This study aimed to determine the concentrations of 20 regulated AAs in clothes marketed in Brazil and Spain. Generally, higher levels of regulated AAs were found in samples sold in Brazil than in Spain, which is linked to the lack of regulation. Sixteen AAs showed concentrations above 5 mg/kg in samples commercialized in Brazil, while 11 exceeded that threshold in Spain. Regulated AAs with levels above 5 mg/kg were more found in synthetic clothes of pink color. Concentrations in clothing were also used to evaluate the dermal exposure to AAs in 3 vulnerable population groups. The highest exposure corresponded to 2,4-diaminoanisole for toddlers in Brazil and 4,4-oxydianiline for newborns in Spain. Non-cancer risks associated with exposure to 4,4-benzidine by Brazilian toddlers was 14.5 (above the threshold). On the other hand, 3,3-dichlorobenzidine was associated with potential cancer risks for newborns and toddlers in Brazil. Given this topic's importance, we recommend conducting continuous studies to determine the co-occurrence of carcinogenic substances.

A simple method to quantify azo dyes in spices based on flow injection chromatography combined with chemometric tools.

Para Red (PR) and Sudan dyes have been illegally used as colorants to adulterate certain foods by enhancing their red/orange colour. In addition, they are toxic and carcinogenic. This work presents the development of a simple flow injection chromatographic method combined with chemometric tools to perform the determination of PR, Sudan I (SI) and Sudan II (SII) in food samples. The flow chromatographic system consisted of a low-pressure manifold coupled to a reverse phase monolithic column. A Partial Least Square (PLS) model was applied to resolve overlapped absorption spectra registered for each dye at the corresponding retention time. The relative errors of calibration (RMSECV, %) were 0.49, 0.85 and 0.23, and the relative errors of prediction (RMSEP, %) were 1.12, 0.75 and 0.33 for PR, SI and SII, respectively. The residual predictive deviation (RPD) values obtained were higher than 3.00 for all analytes. The method was successfully applied to quantify the dyes in six different commercial spices samples. The results were compared with the HPLC reference method concluding that there were no significant differences at the studied confidence level (α = 0.05). The proposed method can be used to rapidly determine the analytes in a simple, reliable, low-cost and environmentally-friendly manner. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05299-8.

Decolorization and detoxification of different azo dyes by Phanerochaete chrysosporium ME-446 under submerged fermentation.

Azo dyes are widely used in the textile industry due to their resistance to light, moisture, and oxidants. They are also an important class of environmental contaminant because of the amount of dye that reaches natural water resources and because they can be toxic, mutagenic, and carcinogenic. Different technologies are used for the decolorization of wastewater containing dyes; among them, the biological processes are the most promising environmentally. The aim of this study was to evaluate the potential of Phanerochaete chrysosporium strain ME-446 to safely decolorize three azo dyes: Direct Yellow 27 (DY27), Reactive Black 5 (RB5), and Reactive Red 120 (RR120). Decolorization efficiency was determined by ultraviolet-visible spectrophotometry and the phytotoxicity of the solutions before and after the fungal treatment was analyzed using Lactuca sativa seeds. P. chrysosporium ME-446 was highly efficient in decolorizing DY27, RB5, and RR120 at 50 mg L-1, decreasing their colors by 82%, 89%, and 94% within 10 days. Removal of dyes was achieved through adsorption on the fungal mycelium as well as biodegradation, inferred by the changes in the dyes' spectral peaks. The intensive decolorization of DY27 and RB5 corresponded to a decrease in phytotoxicity. However, phytotoxicity increased during the removal of color for the dye RR120. The ecotoxicity tests showed that the absence of color does not necessarily translate to an absence of toxicity.

A promising Ames battery for mutagenicity characterization of new dyes.

When testing new products, potential new products, or their impurities for genotoxicity in the Ames test, the quantity available for testing can be a limiting factor. This is the case for a dye repository of around 98,000 substances the Max Weaver Dye Library (MWDL). Mutagenicity data on dyes in the literature, although vast, in several cases is not reliable, compromising the performance of the in silico models. In this report, we propose a strategy for the generation of high-quality mutagenicity data for dyes using a minimum amount of sample. We evaluated 15 dyes from different chemical classes selected from 150 representative dyes of the MWDL. The purity and molecular confirmation of each dye were determined, and the microplate agar protocol (MPA) was used. Dyes were tested at the limit of solubility in single and concentration-response experiments using seven strains without and with metabolic activation except for anthraquinone dyes which were tested with eight strains. Six dyes were mutagenic. The most sensitive was YG1041, followed by TA97a > TA98 > TA100 = TA1538 > TA102. YG7108 as well as TA1537 did not detect any mutagenic response. We concluded that the MPA was successful in identifying the mutagenicity of dyes using less than 12.5 mg of sample. We propose that dyes should be tested in a tiered approach using YG1041 followed by TA97a, TA98, and TA100 in concentration-response experiments. This work provides additional information on the dye mutagenicity database available in the literature.

O estudo teórico do desempenho dos polímeros condutores dos corantes azoicos na detecção eletroquímica de indigo-carmim / The theoretical study of the function of conducting polymers of the azo-dyes in the electrochemical detection of indigo-carmine

RESUMO O desempenho dos polímeros condutores dos corantes azoicos durante a detecção eletroquímica de indigo-carmim foi investigado do ponto de vista teórico, sendo o modelo, correspondente ao caso, descrito e analisado mediante a teoria de estabilidade lineal e da análise de bifiircações. Foi mostrado que o sistema eletroanalítico depende fortemente do pH, pois as concentrações excessivas dos prótons levam à ineficiência eletroanalítica, haja vista o bloqueio dos centros ativos da reação. No entretanto, malgrado o supracitado, os polímeros dos corantes azoicos são modificadores eficientes para determinação do indigo-carmim. A possibilidade das instabilidades oscilatória e monotônica também foi verificada.

SUMMARY The function of the conducting polymers of azo-dyes during the indigo-carmine electrochemical detection has been investigated from the theoretical point of view. The correspondent model has been described and analyzed by means of linear stability theory and bifurcation analysis. It has been shown that the electroanalytical system depends strongly on pH, as the excessive protons concentrations drive the system to the electroanalytical inefficiency, as they block the reaction active sites. Nevertheless, despite of the mentioned, the azo-dyes conducting polymers are efficient modifiers for indigo-carmine electrochemical determination. The possibility of oscillatory and monotonic instabilities has also been verified.

Congo Red Decolorization Using Textile Filters and Laccase-Based Nanocomposites in Continuous Flow Bioreactors.

Removal of azo and diazo dye content from textile industry wastewaters is crucial due to their environmental impact. Here, we report on the use of the fungal laccase from Pycnoporus sanguineus CS43 immobilized on silica nanoparticles and entrapped in textile-based filters for the degradation of Congo Red. Laccase immobilization and synthesis of the nanocomposites were carried out by two different methods, one in the presence of acetone and the second using water as solvent. This led to a change in the hydrophobicity of the obtained biofilters. Successful preparation of the nanocomposites was confirmed via FTIR spectroscopy. Changes in the secondary structure of the enzyme were inspected through the second derivative of the FTIR spectra. Six different types of filter were fabricated and tested in a continuous flow bioreactor in terms of their decolorization capabilities of Congo Red. The results indicate removal efficiencies that approached 40% for enzymes immobilized on the more hydrophobic supports. Backscattered electron (BSE) images of the different filters were obtained before and after the decolorization process. Percentage of decolorization and activity loss were determined as a function of time until a plateau in decolorization activity was reached. Experimental data was used to recreate the decolorization process in COMSOL Multiphysics® (Stockholm, Sweden). These simulations were used to determine the proper combination of parameters to maximize decolorization. Our findings suggest that the treatment of textile-based filters with immobilized laccase in conjunction with hydrophobic nanocomposites provides a suitable avenue to achieve more efficient laccase dye decolorization (39%) than that obtained with similar filters treated only with free laccase (8%). Filters treated with silica-based nanocomposites and immobilized laccases showed an increase in their decolorization capability, probably due to changes in their wetting phenomena.

Characterization of the biosorption of fast black azo dye K salt by the bacterium Rhodopseudomonas palustris 51ATA strain

BACKGROUND: Removal of dyes from wastewater by microorganisms through adsorption, degradation, or accumulation has been investigated. Biological methods used for dye treatment are generally always effective and environmentally friendly. In this study, biosorption of the Fast Black K salt azo dye by the bacterium Rhodopseudomonas palustris 51ATA was studied spectrophotometrically, at various pH (2­10), temperatures (25°C, 35°C, and 45°C) and dye concentrations (25­400 mg L-1). RESULTS: The bacterial strain showed extremely good dye-removing potential at various dye concentrations. IR studies at different temperatures showed that the dye was adsorbed on the bacterial surface at lower temperatures. Characteristics of the adsorption process were investigated by Scatchard analysis at 25°C and 35°C. Scatchard analysis of the equilibrium binding data for the dye on this bacterium gave rise to linear plots, indicating that the Langmuir model could be applied. The regression coefficients obtained for the dye from the Freundlich and Langmuir models were significant and divergence from the Scatchard plot was observed. CONCLUSION: The adsorption behavior of the dye on this bacterium was expressed by the Langmuir, Freundlich, and Temkin isotherms. The adsorption data with respect to various temperatures provided an excellent fit to the Freundlich isotherm. However, when the Langmuir and Temkin isotherm models were applied to these data, a good fit was only obtained for the dye at lower temperatures, thus indicating that the biosorption ability of R. palustris 51ATA is dependent on temperature, pH, and dye concentration.

Magnetic Porous Controlled Fe3O4-Chitosan Nanostructure: An Ecofriendly Adsorbent for Efficient Removal of Azo Dyes.

In this work, chitosan/magnetite nanoparticles (ChM) were quickly synthesized according to our previous report based on co-precipitation reaction under ultrasound (US) irradiation. Besides ChM was in-depth structurally characterized, showing a crystalline phase corresponding to magnetite and presenting a spheric morphology, a "nanorod"-type morphology was also obtained after increasing reaction time for eight minutes. Successfully, both morphologies presented a nanoscale range with an average particle size of approximately 5-30 nm, providing a superparamagnetic behavior with saturation magnetization ranging from 44 to 57 emu·g-1. As ChM nanocomposites have shown great versatility considering their properties, we proposed a comparative study using three different amine-based nanoparticles, non-surface-modified and surface-modified, for removal of azo dyes from aqueous solutions. From nitrogen adsorption-desorption isotherm results, the surface-modified ChMs increased the specific surface area and pore size. Additionally, the adsorption of anionic azo dyes (reactive black 5 (RB5) and methyl orange (MO)) on nanocomposites surface was pH-dependent, where surface-modified samples presented a better response under pH 4 and non-modified one under pH 8. Indeed, adsorption capacity results also showed different adsorption mechanisms, molecular size effect and electrostatic attraction, for unmodified and modified ChMs, respectively. Herein, considering all results and nanocomposite-type structure, ChM nanoparticles seem to be a suitable potential alternative for conventional anionic dyes adsorbents, as well as both primary materials source, chitosan and magnetite, are costless and easily supplied.

Enhanced Catalytic Dye Decolorization by Microencapsulation of Laccase from P. Sanguineus CS43 in Natural and Synthetic Polymers.

Polymeric microcapsules with the fungal laccase from Pycnoporus sanguineus CS43 may represent an attractive avenue for the removal or degradation of dyes from wastewaters. Microcapsules of alginate/chitosan (9.23 ± 0.12 µm) and poly(styrenesulfonate) (PSS) (9.25 ± 0.35 µm) were synthesized and subsequently tested for catalytic activity in the decolorization of the diazo dye Congo Red. Successful encapsulation into the materials was verified via confocal microscopy of labeled enzyme molecules. Laccase activity was measured as a function of time and the initial reaction rates were recovered for each preparation, showing up to sevenfold increase with respect to free laccase. The ability of substrates to diffuse through the pores of the microcapsules was evaluated with the aid of fluorescent dyes and confocal microscopy. pH and thermal stability were also measured for encapsulates, showing catalytic activity for pH values as low as 4 and temperatures of about 80 °C. Scanning electron microscope (SEM) analyses demonstrated the ability of PSS capsules to avoid accumulation of byproducts and, therefore, superior catalytic performance. This was corroborated by the direct observation of substrates diffusing in and out of the materials. Compared with our PSS preparation, alginate/chitosan microcapsules studied by others degrade 2.6 times more dye, albeit with a 135-fold increase in units of enzyme per mg of dye. Similarly, poly(vinyl) alcohol microcapsules from degrade 1.7 times more dye, despite an eightfold increase in units of enzyme per mg of dye. This could be potentially beneficial from the economic viewpoint as a significantly lower amount of enzyme might be needed for the same decolorization level achieved with similar encapsulated systems.

Levedura Pichia kudriavzevii SD5 como biocatalizador na descoloração do corante Preto Reativo 5 / Pichia kudriavzevii yeast as biocatalizer in Reactive Black 5

RESUMO As leveduras vêm apresentando bons resultados na biodegradação de corantes, tornando-se uma alternativa ambientalmente segura e de custo mais baixo para o tratamento de efluentes contendo corantes industriais. Assim, o objetivo deste trabalho foi estudar a descoloração do azo corante Preto Reativo 5 (PR5) pela levedura Pichia kudriavzevii SD5. Para otimização dos parâmetros de descoloração do Preto Reativo 5 foram realizados dois planejamentos experimentais do tipo Delineamento Composto Central Rotacional (DCCR) 23, sendo cada planejamento composto por 17 ensaios, a 150 rpm durante 24 h. As variáveis estudadas foram pH, temperatura e concentração do corante e tiveram como resposta a porcentagem de descoloração. Também foram realizados testes de toxicidade do sobrenadante após 16 h e 24 h de cultivo, utilizando-se sementes de alface (Lactuca sativa) e o microcrustáceo Artemia salina. Os resultados mostraram que a P. kudriavzevii SD5 é uma levedura tolerante a vários tipos de estresse, uma vez que apresentou capacidade de degradar elevadas concentrações do corante PR5 a 45° C. Entretanto, apenas a temperatura apresentou influência estatisticamente significativa (p < 0,05) na descoloração do PR5. Os bioensaios de toxicidade demonstraram que ocorreu diminuição da toxicidade após 24 h de cultivo, e o perfil de absorbância do sobrenadante apontou para um mecanismo degradativo de descoloração.

ABSTRACT Yeasts have been showing good results in the biodegradation of industrial dyes, becoming an environmentally safe and cost-effective alternative for the treatment of effluents containing industrial dyes. Thus, the objective of this work was to study the discoloration of the Reactive Black 5 (RB5) azo dye by the yeast Pichia kudriavzevii SD5. Two experimental designs were employed to optimize the discoloration parameters by means of Central Composite Design (CCD) 23, totalizing 17 trials each, at 150 rpm for 24 h. The studied independent variables were pH, temperature, and concentration of the dye and the outcome parameter was the rate of decolorization (%). Furthermore, the toxicity bioassays of the supernatant after 16 and 24 h of the culture were carried out using lettuce seeds (Lactuca sativa) and the microcrustacean Artemia salina. Results showed that P. kudriavzevii SD5 is a multi-stress tolerant yeast, being capable to degrade high concentrations of RB5 at 45° C. However, only the temperature showed statistical significance (p < 0.05) for dye discoloration. Toxicity bioassays demonstrated that toxicity decreased after 24 h of culture and the absorbance profile of the supernatant pointed to a degradative mechanism of discoloration.

Structural-functional analyses of textile dye degrading azoreductase, laccase and peroxidase: a comparative in silico study

Background: Textile industry not only plays a vital role in our daily life but also a prominent factor in improving global economy. One of the environmental concern is it releases huge quantities of toxic dyes in the water leading to severe environmental pollution. Bacterial laccase and azoreductase successfully oxidize complex chemical structure of nitrogen group-containing azo dyes. Additionally, the presence of textile dye infuriates bacterial peroxidase to act as a dye degrading enzyme. Our present study deals with three textile dye degrading enzymes laccase, azoreductase, and peroxidase through analyzing their structural and functional properties using standard computational tools. Result: According to the comparative analysis of physicochemical characteristics, it was clear that laccase was mostly made up of basic amino acids whereas azoreductase and peroxidase both comprised of acidic amino acids. Higher aliphatic index ascertained the thermostability of all these three enzymes. Negative GRAVY value of the enzymes confirmed better water interaction of the enzymes. Instability index depicted that compared to laccase and preoxidase, azoreductase was more stable in nature. It was also observed that the three model proteins had more than 90% of total amino acids in the favored region of Ramachandran plot. Functional analysis revealed laccase as multicopper oxidase type enzyme and azoreductase as FMN dependent enzyme, while peroxidase consisted of α-ß barrel with additional haem group. Conclusion: Present study aims to provide knowledge on industrial dye degrading enzymes, choosing the suitable enzyme for industrial set up and to help in understanding the experimental laboratory requirements as well.

Azo dyes decolorization under high alkalinity and salinity conditions by Halomonas sp. in batch and packed bed reactor.

Biodecolorization and biodegradation of azo dyes are a challenge due to their recalcitrance and the characteristics of textile effluents. This study presents the use of Halomonas sp. in the decolorization of azo dyes Reactive Black 5 (RB5), Remazol Brilliant Violet 5R (RV5), and Reactive Orange 16 (RO16) under high alkalinity and salinity conditions. Firstly, the effect of air supply, pH, salinity and dye concentration was evaluated. Halomonas sp. was able to remove above 84% of all dyes in a wide range of pH (6-11) and salt concentrations (2-10%). The decolorization efficiency of RB5, RV5, and RO16 was found to be ≥ 90% after 24, 13 and 3 h, respectively, at 50 mg L-1 of dyes. The process was monitored by HPLC-DAD, finding a reduction of dyes along the time. Further, Halomonas sp. was immobilized in volcanic rocks and used in a packed bed reactor for 72 days, achieving a removal rate of 3.48, 5.73, and 8.52 mg L-1 h-1, for RB5, RV5 and RO16, respectively, at 11.8 h. The study has confirmed the potential of Halomonas sp. to decolorize azo dyes under high salinity and alkalinity conditions and opened a scope for future research in the treatment of textile effluents.

Exposure to the azo dye Direct blue 15 produces toxic effects on microalgae, cladocerans, and zebrafish embryos.

Aquatic pollution caused by dyes has increased together with the growth of activities using colorants such as the textile, leather, food, and agrochemicals industries. Because most popular azo dyes are synthesized from benzidine, a carcinogenic compound, a threat to aquatic biota could be expected. The use of single species for toxicity assessment provides limited data, so a battery of test organisms, including representatives of different trophic levels such as algae, zooplankters, and fish, could undoubtedly provide more information. Therefore, our study was aimed at evaluating the toxic effect of the azo dye Direct blue 15 (DB15) on a battery of bioassays using a primary producer (Pseudokirchneriella subcapitata), a primary consumer (Ceriodaphnia dubia), and a secondary consumer (Danio rerio). P. subcapitata was more sensitive to DB15 (IC50 = 15.99 mg L-1) than C. dubia (LC50: 450 mg L-1). In the algae exposed to DB15, chlorophyll-a and -b were significantly increased, and carotenoids were reduced. The concentrations of protein, carbohydrates, and lipids per cell in P. subcapitata exposed to all DB15 concentrations were significantly higher than that measured in control. At 25 mg L-1 of DB15, survival, total progeny, and the number of released clutches were significantly decreased, and the start of reproduction was delayed in C. dubia. DB15 did not induce lethal or sublethal effects in D. rerio embryos at any of the tested concentrations from 24 to 72 h post-fertilization (hpf), but from 96 to 144 hpf, the larvae exposed to 100 and 500 mg L-1 developed yolk sac edema, curved tail, and skeletal deformations. After 144 hpf, DB15 produced a significant increase in embryos without a heartbeat, as the concentration of dye raised. The textile-used, azo dye DB15, caused toxic effects of different magnitude on microalgae, cladocerans, and zebrafish embryos; for this reason, the discharge of this colorant into waterbodies should be regulated to prevent environmental impacts.