Cloning and characterization of FMN-dependent azoreductases from textile industry effluent identified through metagenomic sequencing.

Azo dyes, when released untreated in the environment, cause detrimental effects on flora and fauna. Azoreductases are enzymes capable of cleaving commercially used azo dyes, sometimes in less toxic by-products which can be further degraded via synergistic microbial cometabolism. In this study, azoreductases encoded by FMN1 and FMN2 genes were screened from metagenome shotgun sequences generated from the samples of textile dye industries' effluents, cloned, expressed, and evaluated for their azo dye decolorization efficacy. At pH 7 and 45°C temperature, both recombinant enzymes FMN1 and FMN2 were able to decolorize methyl red at 20 and 100 ppm concentrations, respectively. FMN2 was found to be more efficient in decolorization/degradation of methyl red than FMN1. This study offers valuable insights into the possible application of azoreductases to reduce the environmental damage caused by azo dyes, with the hope of contributing to sustainable and eco-friendly practices for the environment management. This enzymatic approach offers a promising solution for the bioremediation of textile industrial effluents. However, the study acknowledges the need for further process optimization to enhance the efficacy of these enzymes in large-scale applications.Implications: The study underscores the environmental hazards associated with untreated release of azo dyes into the environment and emphasizes the potential of azoreductases, specifically those encoded by FMN1 and FMN2 genes, to mitigate the detrimental effects. The study emphasizes the ongoing commitment to refining and advancing the enzymatic approach for the bioremediation of azo dye-containing effluents, marking a positive stride toward more sustainable industrial practices.

Biotransformation of the azo dye reactive orange 16 by Aspergillus flavus A5P1: Performance, genetic background, pathway, and mechanism.

This study reports the strain Aspergillus flavus A5P1 (A5P1), which is with the capable of degrading the azo dye reactive orange 16 (RO16). The mechanism of RO16 degradation by A5P1 was elucidated through genomic analysis, enzymatic analysis, degradation pathway analysis and oxidative stress analysis. Strain A5P1 exhibited aerobic degradation of RO16, with optimal degradation at an initial pH of 3.0. Genomic analysis indicates that strain A5P1 possesses the potential for acid tolerance and degradation of azo dye. Enzymatic analysis, combined with degradation product analysis, demonstrated that extracellular laccase, intracellular lignin peroxidase, and intracellular quinone reductase were likely key enzymes in the RO16 degradation process. Oxidative stress analysis revealed that cell stress responses may participate in the RO16 biotransformation process. The results indicated that the biotransformation of RO16 may involves biological processes such as transmembrane transport of RO16, cometabolism of the strain with RO16, and cell stress responses. These findings shed light on the biodegradation of RO16 by A5P1, indicating A5P1's potential for environmental remediation.

Promotion of anaerobic biodegradation of azo dye RR2 by different biowaste-derived biochars: Characteristics and mechanism study by machine learning.

The addition of biochar resulted in a 31.5 % to 44.6 % increase in decolorization efficiency and favorable decolorization stability. Biochar promoted extracellular polymeric substances (EPS) secretion, especially humic-like and fulvic-like substances. Additionally, biochar enhanced the electron transfer capacity of anaerobic sludge and facilitated surface attachment of microbial cells. 16S rRNA gene sequencing analysis indicated that biochar reduced microbial species diversity, enriching fermentative bacteria such as Trichococcus. Finally, a machine learning model was employed to establish a predictive model for biochar characteristics and decolorization efficiency. Biochar electrical conductivity, H/C ratio, and O/C ratio had the most significant impact on RR2 anaerobic decolorization efficiency. According to the results, the possible mechanism of RR2 anaerobic decolorization enhanced by different types of biochar was proposed.

Improved application of immobilized Enterobacter cloacae into a bio-based polymer for Reactive Blue 19 removal, an eco-friendly advancement in potential decolorizing systems.

The widespread use of highly complex synthetic dyes like reactive dyes in the textile industry has some adverse environmental impacts and deserves close attention. Biological treatment of these effluents utilizing various species of bacteria with remarkable efficiency in dye removal is still considered promising. Our current study deals with immobilizing an isolated bacterial strain into calcium alginate (Ca/Alg) gel beads and using it to treat pernicious pollutants like synthetic dyes. A potential Reactive Blue 19 (RB19)-degrading Enterobacter cloacae strain A1 was isolated from the Kashan textile industry and was characterized by 16S rDNA gene sequencing. The decolorization ability of strain A1 was assessed by time-based studies using free bacterial cells/immobilized in Ca/Alg. Based on the results of the 16S rDNA gene sequencing, it appears that strain A1 belonged to E. cloacae, with a 99.74% similarity. The findings suggest that immobilized strain A1 accomplished maximum decolorization activity compared with the free cells. The immobilized strain could utterly decompose and decolorize 0.05 mg/mL of RB19 within 48 h, while the free bacterial strain decolorized RB19 within 5 days. Moreover, Ca/Alg gel beads can maintain their efficiency for over three decolorization cycles. Further infrared spectroscopy (FTIR) and gas chromatograph mass spectrometer (GC/MS) investigation declared complete RB19 decomposition on reaction products. Artemia salina was used to investigate the toxicity of dye and its degraded metabolites. The LC50 values signified the pure dye as very toxic with 0.01 mg/mL concentration, while after-treatment products showed no toxic effect on larvae. This immobilization technique increased the applicability of bacterial strain for dye removal. It was beneficial for the decolorization of RB19 from textile wastewater due to a remarkable reduction in time. Notably, strain A1-immobilized beads can maintain their activity for three consecutive decolorization cycles without a considerable decrease in efficiency. PRACTITIONER POINTS: The remarkable capacity of immobilized Enterobacter cloacae strain A1 for Reactive Blue 19 (RB19) removal Immobilized A1 strain showed two-fold higher removal than free one over 48 h A promising method for enhancing RB19 decolorization Decolorization was due to degradation based on UV-Vis, FTIR, and GC/MS analysis Non-toxic posttreatment products for Artemia.

Biodegradation of commercial textile reactive dye mixtures by industrial effluent adapted bacterial consortium VITPBC6: a potential technique for treating textile effluents.

Textile industries release major fraction of dyestuffs in effluents leading to a major environmental concern. These effluents often contain more than one dyestuff, which complicates dye degradation. In this study ten reactive dyes (Reactive Yellow 145, Reactive Yellow 160, Reactive Orange 16, Reactive Orange 107, Reactive Red 195, Reactive Blue 21, Reactive Blue 198, Reactive Blue 221, Reactive Blue 250, and Reactive Black 5) that are used in textile industries were subjected to biodegradation by a bacterial consortium VITPBC6, formulated in our previous study. Consortium VITPBC6 caused single dye degradation of all the mentioned dyes except for Reactive Yellow 160. Further, VITPBC6 efficiently degraded a five-dye mixture (Reactive Red 195, Reactive Orange 16, Reactive Black 5, Reactive Blue 221, and Reactive Blue 250). Kinetic studies revealed that the five-dye mixture was decolorized by VITPBC6 following zero order reaction kinetic; Vmax and Km values of the enzyme catalyzed five-dye decolorization were 128.88 mg L-1 day-1 and 1003.226 mg L-1 respectively. VITPBC6 degraded the dye mixture into delta-3,4,5,6-Tetrachlorocyclohexene, sulfuric acid, 1,2-dichloroethane, and hydroxyphenoxyethylaminohydroxypropanol. Phytotoxicity, cytogenotoxicity, microtoxicity, and biotoxicity assays conducted with the biodegraded metabolites revealed that VITPBC6 lowered the toxicity of five-dye mixture significantly after biodegradation.

Biodegradation and reduction of toxicity of Azo Trypan Blue dye by Amazonian strains of gasteroid fungi (Basidiomycota).

Amazonian strains of Cyathus spp. and Geastrum spp. were studied for the ability to discolor the trypan blue azo dye and reduce its toxicity. Discoloration of trypan blue dye (0.05%) was evaluated in solid and aqueous medium over different periods. The reduction of dye toxicity after treatment was assessed by seed germination and the development of lettuce seedlings (Lactuca sativa L.) and toxicity test in Artemia salina (L.) larvae. All evaluated strains showed the potential to reduce the color intensity of trypan blue dye. Cyathus strains reached 96% discoloration, and C. albinus and C. limbatus also reduced dye toxicity. Geastrum strains showed a high efficiency degree in color reduction, reaching 98% discoloration, however, the by-products generated during the process presented toxicity and require further investigation. For the first time, Amazonian strains of gasteroid fungi degrading trypan blue are reported, some even reducing its toxicity. Thus, making them promising sources of enzymes of interest to bioremediation scenarios involving synthetic dyes.

Metabolism of azo food dyes by bacterial members of the human gut microbiome.

OBJECTIVES: We set out to survey the capacities of bacterial isolates from the human gut microbiome to reduce common azo food dyes in vitro. METHODS: A total of 206 strains representative of 124 bacterial species and 6 phyla were screened in vitro using a simple azo dye decolorization assay. Strains which showed azoreductive activity were characterized by studies of azoreduction kinetics and bacterial growth. RESULTS: Several groups of gut bacteria, including ones not previously associated with azoreduction, reduced one or more of the four azo food dyes commonly used in Canada: Allura Red, Amaranth, Sunset Yellow, and Tartrazine. Strains within some species differed in their azoreductive capabilities. Some strains displayed evidence of effects on growth related to the presence of azo dyes and/or the products of their azoreduction. CONCLUSION: The continued widespread use of food azo dyes requires re-evaluation in light of the potential for disturbance of the gut microbial ecosystem resulting from azoreduction and the possibility of consequences for human health.

Degradation and decolorization of textile azo dyes by effective fungal-bacterial consortium.

BACKGROUND: Synthetic dyes are one of the main pollutants in the textile industry and bioremediation is considered as an environmentally friendly method to degrade them. Soil microbial consortia (MCs) are reported having the potential of decolorizing most of textile dyes. This study aimed at evaluating dye-degrading ability of MCs developed from fungi and bacteria. METHODS AND RESULTS: Fungi and bacteria were isolated from the soil samples obtained from textile waste dumping site at Horana industrial zone, Sri Lanka and were screened for crystal violet (CV) and congo red (CR) dyes to develop MCs. Decolorization assay was performed for MCs along with individual isolates under variable pH levels. Metabolized products were characterized to confirm the biodegradation. A. tamari (F5) and P. putida (B3) significantly (P < 0.05) decolorized both dyes. All the MCs showed higher decolorization percentages over the individual microorganisms. Neutral pH (pH 7) was the optimum pH for the decolorization of both dyes by individual isolates and the percentages were significantly changed under the acidic and basic pH levels. However, decolorization ability by all MCs was not significantly changed with pH. Consortium with A. tamari - P. putida significantly (P < 0.05) decolourized both dyes under optimum pH 7. CONCLUSION: All MCs showed better pH tolerance in degrading CV and CR. Thus, it can be concluded that the selected MC with A. tamari - P. putida can degrade CV and CR textile dyes efficiently into non-toxic compounds against plants under neutral pH. Degradation and decolorization of textile azo dyes by effective fungal-bacterial consortium.

The global anaerobic metabolism regulator fnr is necessary for the degradation of food dyes and drugs by Escherichia coli.

IMPORTANCE: This work has broad relevance due to the ubiquity of dyes containing azo bonds in food and drugs. We report that azo dyes can be degraded by human gut bacteria through both enzymatic and nonenzymatic mechanisms, even from a single gut bacterial species. Furthermore, we revealed that environmental factors, oxygen, and L-Cysteine control the ability of E. coli to degrade azo dyes due to their impacts on bacterial transcription and metabolism. These results open up new opportunities to manipulate the azoreductase activity of the gut microbiome through the manipulation of host diet, suggest that azoreductase potential may be altered in patients suffering from gastrointestinal disease, and highlight the importance of studying bacterial enzymes for drug metabolism in their natural cellular and ecological context.

Enhanced Biodegradation of Methyl Orange Through Immobilization of Shewanella oneidensis MR-1 by Polyvinyl Alcohol and Sodium Alginate.

Shewanella oneidensis MR-1 has great potential for use in remediating azo dye pollution. Here, a new high-efficiency biodegradation method was developed utilizing S. oneidensis MR-1 immobilized by polyvinyl alcohol (PVA) and sodium alginate (SA). After determining the optimal immobilization conditions, the effects of various environmental factors on methyl orange (MO) degradation were analyzed. The biodegradation activity of the immobilized pellets was evaluated by analyzing the MO removal efficiency, and characterization was performed using scanning electron microscopy. The MO adsorption kinetics can be described using pseudo-second-order kinetics. Compared with free bacteria, the MO degradation rate of the immobilized S. oneidensis MR-1 increased from 41% to 92.6% after 21 days, suggesting that the immobilized bacteria performed substantially better and had more stable removal rates. These factors indicate the superiority of bacteria entrapment in addition to its easy application. This study demonstrates that the application of immobilized S. oneidensis MR-1 entrapped by PVA-SA can be used to establish a reactor with stable and high MO removal rates.

Molecular characterization of azoreductase and its potential for the decolorization of Remazol Red R and Acid Blue 29.

Azoreductase is a reductive enzyme that efficiently biotransformed textile azo dyes. This study demonstrated the heterologous overexpression of the azoreductase gene in Escherichia coli for the effective degradation of Remazol Red-R and Acid-Blue 29 dyes. The AzK gene of Klebsiella pneumoniae encoding a ≈22 kDa azoreductase enzyme was cloned into the pET21+C expression vector. The inoculum size of 1.5%, IPTG concentration of 0.5 mM, and incubation time of 6 h were optimized by response surface methodology a statistical tool. The crude extract showed 76% and 74%, while the purified enzyme achieved 94% and 93% decolorization of RRR and AB-29, respectively in 0.3 h. The reaction kinetics showed that RRR had a Km and Vmax value of 0.058 mM and 1416 U mg-1, respectively at an NADH concentration of 10 mM. HPLC and GC-MS analyses showed that RRR was effectively bio-transformed by azoreductase to 2-[3-(hydroxy-amino) benzene-1-sulfonyl and AB-29 to aniline and 3-nitrosoaniline. This study explored the potential of recombinant azoreductase isolated from K. pneumoniae in the degradation of toxic textile azo dyes into less toxic metabolites.

Decolorization of reactive azo dye using novel halotolerant yeast consortium HYC and proposed degradation pathway.

The presence of high salinity levels in textile wastewater poses a significant obstacle to the process of decolorizing azo dyes. The present study involved the construction of a yeast consortium HYC, which is halotolerant and was recently isolated from wood-feeding termites. The consortium HYC was mainly comprised of Sterigmatomyces halophilus SSA-1575 and Meyerozyma guilliermondii SSA-1547. The developed consortium demonstrated a decolourization efficiency of 96.1% when exposed to a concentration of 50 mg/l of Reactive Black 5 (RB5). The HYC consortium significantly decolorized RB5 up to concentrations of 400 mg/l and in the presence of NaCl up to 50 g/l. The effects of physicochemical factors and the degradation pathway were systematically investigated. The optimal pH, salinity, temperature, and initial dye concentration were 7.0, 3%, 35 °C and 50 mg/l, respectively. The co-carbon source was found to be essential, and the addition of glucose resulted in a 93% decolorization of 50 mg/l RB5. The enzymatic activity of various oxido-reductases was assessed, revealing that NADH-DCIP reductase and azo reductase exhibited greater activity in comparison to other enzymes. UV-Visible (UV-vis) spectrophotometry, Fourier-transform infrared spectroscopy (FTIR), high-performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC-MS) were utilized to identify the metabolites generated during the degradation of RB5. Subsequently, a metabolic pathway was proposed. The confirmation of degradation was established through alterations in the functional groups and modifications in molecular weight. The findings indicate that this halotolerant yeast consortium exhibits promising potential of degrading dye compounds. The results of this study offer significant theoretical basis and crucial perspectives for the implementation of halotolerant yeast consortia in the bioremediation of textile and hypersaline wastewater. This approach is particularly noteworthy as it does not produce aromatic amines.

Electrogenic engineered flow through tri-phasic wetland system for azo dye treatment: Microbial dynamics and functional metagenomics.

Electrogenic engineered flow through tri-phasic wetland (EEFW) system based on nature-based ecological principles was studied by integrating successive biological microenvironments. The potential mechanism of the plant root-based microbial community and its functional diversity with the influence of plant-microbe-electrode synergism towards dye degradation was evaluated. The EEFW system was operated at three varied dye loads of 10, 25 and 50 mg L-1, where the results from the cumulative outlets revealed a maximum dye removal efficiency of 96%, 96.5% and 93%, respectively. Microbial community analysis depicted synergistic dependence on the plant-microbe-electrode interactions, influencing their functional diversity and metabolism towards detoxification of pollutants. The core microbial taxa enriched against the microenvironment variation were mostly associated with carbon and dye removal viz., Desulfomonile tiedjei and Rhodopseudomonas palustris in Tank 1 and Chloroflexi bacterium and Steroidobacter denitrificans in Tank 2. The degradation of polycyclic aromatic hydrocarbons, chloroalkane/chloroalkene, nitrotoluene, bisphenol, caprolactam and 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) were observed to be predominant in Tank 1. EEFW system could be one of the option for utilizing nature-based processes for the treatment of wastewater by self-induced bioelectrogenesis to augment process efficiency.

Enzymatic degradation of azo dyes methylene blue and congo red with peroxidase purified from cauliflower using affinity chromatography technique: Kinetic study, optimization and metal binding activity.

The effective results of the enzymatic decolorization of industrial azo dyes found in wastewater, which cause serious health and environmental problems, with peroxidases have recently increased the interest in these enzyme sources. Redox-mediated decolorization of Methylene Blue and Congo Red azo dyes with cauliflower (Brassica oleracea var. botrytis L.) peroxidase (CPOD) purified in one step using 4-amino 3-bromo 2-methyl benzohydrazide molecule was investigated for the first time. The inhibition effect of this molecule, which is used as a ligand in affinity chromatography, on the CPOD enzyme was investigated. The Ki and IC50 values for this enzyme were calculated as 0.113 ± 0.012 mM and 0.196 ± 0.011 mM, respectively. With the affinity gel obtained by binding to the Sepharose-4B-l-tyrosine matrix of this molecule, which shows a reversible inhibition effect, the purification values of CPOD enzyme were determined as 562-fold with a specific activity of 50,250 U mg-1. The purity of the enzyme was checked by the SDS-PAGE technique and its molecular weight was determined. A single band at 44 kDa was observed for the CPOD enzyme. In dye decolorization studies, the effects of dye, enzyme, and hydrogen peroxide concentrations as well as time, pH, and temperature were investigated. The profiles of the optimum conditions for both dyes were similar, and the percentages of decolorization of Methylene Blue and Congo Red under these conditions were 89% and 83%, respectively, at the end of the 40 min reaction time. Again, when examining the effect of metal ions on enzyme activity, it was found that there was no significant negative change in CPOD.

Nanobioremediation: a bacterial consortium-zinc oxide nanoparticle-based approach for the removal of methylene blue dye from wastewater.

Industrial effluents carrying dyes are considered a major environmental threat in the present era. Methylene blue (MB) dye is one of the key dyes of the thiazine group of dyes. It is broadly used in medical, textile, and various fields and is well known for its carcinogenicity and methemoglobin nature. Bacterial and other microbes-mediated bioremediation is becoming an emerging and significant section for the treatment of wastewater. Isolated bacteria were used for the bioremediation and nanobioremediation of methylene blue dye under varying conditions and parameters. A comparative study was conducted for the remediation of methylene blue dye using bacterial consortium, potential bacteria (isolated by scale-up method), and potential bacteria within zinc oxide nanoparticles. The decolorizing ability of bacteria was analyzed by UV visible spectrophotometer after stirring and static incubation in different time intervals of the isolates. Growth parameters and environmental parameters which include pH, initial dye concentration, and dose of nanoparticles were optimized with the minimal salt medium. An enzyme assay study was also done to check the effect of dye and nanoparticles on bacterial growth and the mode of action of degradation. The authors found that potential bacteria within ZnO nanoparticles showed enhanced decolorization efficiency (95.46% at pH 8) due to the properties of nanoparticles. On the other hand, the decolorization of MB dye by potential bacteria and the bacterial consortium was about 89.08 and 76.3%, respectively, for a 10-ppm dye concentration. During the enzyme assays study, the highest activity was observed for phenol oxidase, nicotinamide adenine dinucleotide (NADH), 2,6-Dichloroindophenol(DCIP), and laccase for nutrient broth having MB dye, MB dye, and ZnO NPs, while no such change was observed for manganese peroxidase enzyme activity. Nanobioremediation is a promising approach to removing such pollutants from the environment.

Simultaneous removal of refractory organic pollutants and nitrogen using electron shuttle suspended biofilm carriers in an integrated hydrolysis/acidification-anoxic/aerobic process.

Azo dyes wastewater contains refractory pollutant and nitrogen, which threatens human health and ecological environment when discharged into environment directly. Electron shuttle (ES) is able to participate in the extracellular electron transfer, and thus enhances the removal efficiency of refractory pollutant. However, the continuous dosing of soluble ES would rise operation cost and cause contamination inevitably. In this study, a type of insoluble ES (carbonylated graphene oxide (C-GO)) was developed and melt blended into polyethylene (PE) to prepare novel C-GO-modified suspended carriers. Compared to those of conventional carrier (31.60%), the surface active sites of novel C-GO-modified carrier increased to 52.95%. An integrated hydrolysis/acidification (HA, filled with C-GO-modified carrier) - anoxic/aerobic (AO, filled with clinoptilolite-modified carrier) process was applied to remove azo dye acid red B (ARB) and nitrogen simultaneously. ARB removal efficiency was significantly improved in the reactor filled with C-GO-modified carriers (HA2) compared to the reactor filled with conventional PE carriers (HA1) or activated sludge (HA0). Total nitrogen (TN) removal efficiency of the proposed process increased by 25.95-32.64% compared to the reactor filled with activated sludge. Moreover, the intermediates of ARB were identified by liquid chromatograph-mass spectrometer (LC-MS), and the degradation pathway of ARB through ES was proposed. C-GO-modified carriers induced ARB-removal-related bacterial enrichment (such as Chloroflexi, Lactivibrio, Longilinea, Bacteroidales and Anaerolineaceae). Besides, the relative abundance of denitrifiers and nitrifiers in the AO reactor filled with clinoptilolite-modified carrier was increased by 11.60% compared with activated sludge. Copy numbers of genes related to membrane transport, carbon/energy metabolism and nitrogen metabolism increased significantly on the surface-modified carriers. This study proposed an efficient approach for simultaneous azo dyes and nitrogen removal, showing potential in actual application.

In situ treatment of real textile effluent in constructed furrows using consortium of Canna indica and Saccharomyces cerevisiae and subsequent biochemical and toxicity evaluation.

Emerging contaminants removals like dyes and heavy metals from the textile effluent have an immense challenge. The present study focuses on the biotransformation and detoxification of dyes and in situ textile effluent treatment by plants and microbes efficiently. A mixed consortium of perennial herbaceous plant Canna indica and fungi Saccharomyces cerevisiae showed decolorization of di-azo dye Congo red (CR, 100 mg/L) up to 97% within 72 h. Root tissues and Saccharomyces cerevisiae cells revealed induction of various dye-degrading oxidoreductase enzymes such as lignin peroxidase, laccase, veratryl alcohol oxidase and azo reductase during CR decolorization. Chlorophyll a, Chlorophyll b and carotenoid pigments were notably elevated in the leaves of a plant during the treatment. Phytotransformation of CR into its metabolic constituents was detected by using several analytical techniques, including FTIR, HPLC, and GC-MS and its non-toxic nature was confirmed by cyto-toxicological evaluation on Allium cepa and on freshwater bivalves. Mix consortium of plant Canna indica and fungi Saccharomyces cerevisiae efficiently treated textile wastewater (500 L) and reduced ADMI, COD, BOD, TSS and TDS (74, 68, 68, 78, and 66%) within 96 h. In situ textile wastewater treatment for in furrows constructed and planted with Canna indica, Saccharomyces cerevisiae and consortium-CS within 4 days reveals reduced ADMI, COD, BOD, TDS and TSS (74, 73, 75, 78, and 77%). Comprehensive observations recommend this is an intelligent tactic to exploit this consortium in the furrows for textile wastewater treatment.

Biotreatment of azo dye containing textile industry effluent by a developed bacterial consortium immobilised on brick pieces in an indigenously designed packed bed biofilm reactor.

This study highlights the development of a lab-scale, indigenously designed; Packed-Bed Biofilm Reactor (PBBR) packed with brick pieces. The developed biofilm in the reactor was used for the decolourisation and biodegradation of the textile industry effluent. The PBBR was continuously operated for 264 days, during which 301 cycles of batch and continuous treatment were operated. In batch mode under optimised conditions, more than 99% dye decolourisation and ≥ 92% COD reduction were achieved in 6 h of contact time upon supplementation of effluent with 0.25 g L-1 glucose, 0.25 g L-1 urea, and 0.1 g L-1 phosphates. A decolourisation rate of 133.94 ADMI units h-1 was achieved in the process. PBBR, when operated in continuous mode, showed ≥ 95% and ≥ 92% reduction in ADMI and COD values. Subsequent aeration and passage through the charcoal reactor assisted in achieving a ≥ 96% reduction in COD and ADMI values. An overall increase of 81% in dye-laden effluent decolourisation rate, from 62 to 262 mg L-1 h-1, was observed upon increasing the flow rate from 18 to 210 mL h-1. Dye biodegradation was determined by UV-Vis and FTIR spectroscopy and toxicity study. SEM analysis showed the morphology of the attached-growth biofilm.

Decoding whole genome of Anoxybacillus rupiensis TPH1 isolated from tatapani hot spring, India and giving insight into bioremediation ability of TPH1 via heavy metals and azo dyes.

A moderately thermophilic, gram-positive genomospecies Anoxybacillus rupiensis TPH1 was isolated from Tatapani hot spring, Chhattisgarh, India. Genome of 3.70 Mb with 42.3% GC subsumed 4131 CDSs, 65 tRNA, 5 rRNA, 35 AMR and 19 drug target genes. Further, comparative genomics of 19 Anoxybacillus spp. exhibited an open pan genome of 13102 genes along with core (10.62%), unique (43.5%) and accessory (45.9%) genes. Moreover, phylogenomic tree displayed clustering of Anoxybacillus spp. into two distinct clades where clade A species harbored larger genomes, more unique genes, CDS and hypothetical proteins than clade B species. Further, distribution of azoreductases showed FMN-binding NADPH azoreductase (AzoRed1) presence in clade A species only and FMN-binding NADH azoreductase (AzoRed2) harboring by species of both clades. Heavy metal resistance genes distribution showed omnipresence of znuA, copZ and arsC in both clades, dispersed presence of cbiM, czcD, merA and feoB over both clades and harboring of nikA and acr3 by few species of clade A only. Additionally, molecular docking of AzoRed1, AzoRed2, ZnuA, CopZ, Acr3, CbiM, CzcD, MerA and NikA with their respective ligands indicated high affinity and stable binding. Conclusively, present study provided insight into gene repertoire of genus Anoxybacillus and a basis for the potential application of this thermophile in bioremediation of azo dyes and heavy metals.

Impact of immobilized algae and its consortium in biodegradation of the textile dyes.

In biological engineering, cell immobilization is a modern technique for immobilizing free cells in a small space. Disintegration and elimination of azo dyes [Reactive Orange 122 (orange 2RL) and Reactive Red 194 (Reactive Red M-2BF)] were investigated by using Chlorococcum sp. and Chlorococcum sp. mixed with Scenedesmus obliquus, respectively. After 7 days of incubation, the maximum decolorization was spotted at 40 ppm for Reactive Orange 122 and 20 ppm for Reactive Red 194 by Chlorococcum sp. and Chlorococcum sp. mixed with S. obliquus, respectively. The findings revealed that the best decolorization activity was found at pH 11 and 25 °C under aeration conditions. BG11 was considered the best medium for azo dye decolorization with a high decolorization percentage. Additionally, different concentrations of nitrogen and phosphorus show the high activity of decolorization of both dyes. Referring to vitamins (thiamin and Ascorbic acid), all studied concentrations showed high decolorization activity with immobilized Chlorococcum sp. mixed with S. obliquus; however, different concentrations (20, 40, and 60 mg/l) of thiamin showed completely decolorization of Reactive Red 194 after 3 days, and 60 mg/l of ascorbic acid showed completely decolorization of Reactive Orange 122 after 5 days of inoculation. FT-IR and GC-Ms analysis for azo dyes after and before treatment with Immobilization of Chlorococcum sp. and Chlorococcum sp. mixed with Scenedesmus obliquus were detected. Novelty statement: The natural carrier algae and its consortium combined with a suitable immobilization technique were considered in this study, which is non-toxic, enhanced their bioremediation potential for dyes, and allowed multiple uses of biocatalysts. The novel use of the immobilization and its consortium of algae on the degradation efficiency of azo dyes and studying the effect of physicochemical conditions on decolorization and degradation of azo dyes. Application of immobilization techniques using microalgae could be excellent bioremediation of wastewaters.