Azoreductase Research Articles

Streptomyces as a Novel Biotool for Azo Pigments Remediation in Contaminated Scenarios.

Azo pigments are widely used in the textile and leather industry, and they generate diverse contaminants (mainly in wastewater effluents) that affect biological systems, the rhizosphere community, and the natural activities of certain species. This review was performed according to the Systematic Reviews and Meta Analyses (PRISMA) methodology. In the last decade, the use of Streptomyces species as biological azo-degraders has increased, and these bacteria are mainly isolated from mangroves, dye-contaminated soil, and marine sediments. Azo pigments such as acid orange, indigo carmine, Congo red, and Evans blue are the most studied compounds for degradation, and Streptomyces produces extracellular enzymes such as peroxidase, laccase, and azo reductase. These enzymes cleave the molecule through asymmetric cleavage, followed by oxidative cleavage, desulfonation, deamination, and demethylation. Typically, some lignin-derived and phenolic compounds are used as mediators to improve enzyme activity. The degradation process generates diverse compounds, the majority of which are toxic to human cells and, in some cases, can improve the germination process in some horticulture plants. Future research should include analytical methods to detect all of the molecules that are generated in degradation processes to determine the involved reactions. Moreover, future studies should delve into consortium studies to improve degradation efficiency and observe the relationship between microorganisms to generate scale-up biotechnological applications in the wastewater treatment industry.

Design and Synthesis of an Azo Reductase Responsive Flavonol-Indomethacin Hybrid Used for the Diagnosis and Treatment of Colitis.

Human intestinal bacteria are the primary producers of azo reductase, and the content of azo reductase is closely associated with various intestinal diseases, including ulcerative colitis (UC). The rapid detection of changes in azo reductase levels is crucial for diagnosing and promptly intervening in UC. In this study, a therapeutic agent, FAI, specifically targeting UC, was designed and synthesized. This agent was developed by linking the anti-inflammatory drug indomethacin to flavonols with antioxidant activity via an azo bond (off-on). Breakage of the azo bond breaks results in the release of both fluorophores and drugs, achieving targeted tracing and integrated treatment effects. In vivo and in vitro fluorescence imaging experiments were used to demonstrate the potential of FAI in the diagnosis of UC, together with synergistic therapeutic effects through the release of both fluorophores and anti-inflammatory agents. Therefore, this diagnostic agent shows promise as a potential tool for diagnosing and treating UC.

Photodynamic therapy promotes hypoxia‐activated nitrogen mustard drug release

AbstractPhotodynamic therapy (PDT) has become a promising method for tumor treatment due to its non‐invasive and high spatiotemporal selectivity. However, PDT is still hindered by reactive oxygen species deficiency, because solid tumors feature a hypoxic microenvironment. PDT combined with hypoxia‐activated chemotherapy drugs can effectively induce tumor death, overcoming the limitations of the sole PDT for the fight against hypoxia. Herein, we designed a nanosystem (PCe6AZOM) that enhances the release of hypoxia‐activated drugs (AZOM) by PDT. Under hypoxic conditions, the azo bond of AZOM is cleaved by azo reductase, releasing highly cytotoxic AZOM and resulting in a significant increase in intratumor drug concentration. Meanwhile, the commercial photosensitizer Ce6 can aggravate the oxygen‐poor state during the PDT process and further cause more AZOM release. Moreover, the cascade reactions in the nanosystem could activate singlet oxygen and enhance drug release through 660 nm light laser irradiation, contributing to more effective induction of tumor apoptosis and tumor growth retardation in vitro and in vivo.

Molecular mechanisms of selenite reduction by Lactiplantibacillus plantarum BSe: An integrated genomic and transcriptomic analysis

The reduction of selenite [Se(Ⅳ)] by microorganisms is a green and efficient detoxification strategy. We found that Se(Ⅳ) inhibited exopolysaccharide and protein secretion by Lactiplantibacillus plantarum BSe and compromised cell integrity. In this study, L. plantarum BSe reduced Se(Ⅳ) by increasing related enzyme activity and electron transfer. Genomic analysis demonstrated that L. plantarum BSe should be able to reduce Se(Ⅳ). Further transcriptome analysis showed that L. plantarum BSe enhanced its tolerance to Se(Ⅳ) by upregulating the expression of surface proteins and transporters, thus reducing the extracellular Se(Ⅳ) concentration through related enzymatic reactions and siderophore-mediated pathways. Lactiplantibacillus plantarum BSe was able to regulate the expression of related genes involved in quorum sensing and a two-component system and then select appropriate strategies for Se(Ⅳ) transformation in response to varying environmental Se(Ⅳ) concentrations. In addition, azo reductase was linked to the reduction of Se(Ⅳ) for the first time. The present study established a multipath model for the reduction of Se(Ⅳ) by L. plantarum, providing new insights into the biological reduction of Se(Ⅳ) and the biogeochemical cycle of selenium.

Remotely Controllable Supramolecular Nanomedicine for Drug-Resistant Colorectal Cancer Therapy Caused by Fusobacterium nucleatum.

Fusobacterium nucleatum (Fn) existing in the community of colorectal cancer (CRC) promotes CRC progression and causes chemotherapy resistance. Despite great efforts that have been made to overcome Fn-induced chemotherapy resistance by co-delivering antibacterial agents and chemotherapeutic drugs, increasing the drug-loading capacity and enabling controlled release of drugs remain challenging. In this study, a novel supramolecular upconversion nanoparticle (SUNP) is constructed by incorporating a positively charged polymer (PAMAM-LA-CD) with Fn inhibition capacity, a negatively charged platinum (IV) oxaliplatin prodrug (OXA-COOH), upconversion nanoparticle (UCNPs) and polyethylene glycol-azobenzene (PEG-Azo) to enhance drug-loading and enable on-demand drug release for drug-resistant CRC treatment. SUNPs exhibit high drug-loading capacity (30.8%) and good structural stability under normal physiological conditions, while disassembled upon exogenous NIR excitation and endogenous azo reductase in the CRC microenvironment to trigger drug release. In vitro and in vivo studies demonstrate that SUNPs presented good biocompatibility and robust performance to overcome chemoresistance, thereby significantly inhibiting Fn-infected cancer cell proliferation. This study leverages multiple dynamic chemical designs to integrate both advantages of drug loading and release in a single system, which provides a promising candidate for precision therapy of bacterial-related drug-resistant cancers.

Azo Reductase Activated Magnetic Resonance Tuning Probe with "Switch-On" Property for Specific and Sensitive Tumor Imaging in Vivo.

Cancer remains a threat to human health. However, if tumors can be detected in the early stage, then the effectiveness of cancer treatment could be significantly improved. Therefore, it is worthwhile to develop more sensitive and accurate cancer diagnostic methods. Herein, we demonstrated an azo reductase (AzoR)-activated magnetic resonance tuning (MRET) probe with a "switch-on" property for specific and sensitive tumor imaging in vivo. Specifically, Gd-labeled DNA1 (DNA1-Gd) and cyclodextrin-coated magnetic nanoparticles (MNP-CD) were employed as enhancer and quencher of MRET, respectively, while DNA2, an azobenzene (Azo) group-modified aptamer (AS1411), served as a linker between enhancer and quencher to construct the MRET probe of MNP@DNA(1-2)-Gd. In tumor tissues with high-level AzoR, the T1-weighted magnetic resonance signal of the MRET probe could be restored by intelligently regulating the switch from "OFF" to "ON" after activation with AzoR, thus accurately indicating the location of the tumor accurately. Moreover, the tumor with a 4 times smaller size than that of the normal tumor model could be imaged based on the proposed MRET probe. The as-proposed MRET-based magnetic resonance imaging strategy not only achieves tumor imaging accurately but also shows promise for early diagnosis of tumors, which might improve patients' survival rates and provide an opportunity for image-guided biomedical applications in the future.

Degradation of synthetic azo dyes by Citrobacter freundii isolated from tannery effluent

AbstractScarcity of water is becoming a big issue. Most of the water is used in industrial sectors including pharmaceutical, food, textiles, leather, and cosmetics. Industrial effluent water is unable to treat and reuse due to their intricate nature. Effluent water contains synthetic azo dyes since they employ synthetic azo dyes as a major coloring agent. Existing approaches are ineffective in treating effluent containing synthetic azo dyes. Synthetic azo dyes containing effluent water can be treated biologically to overcome the drawbacks of physical and chemical processes. In this research work, novel organism was isolated from tannery effluent water. Isolated organisms were screened and selected for effective degradation of azo dyes. One such organism is Citrobacter freundii which effectively degrade the effluent water containing synthetic azo dyes and allowing it to reuse for variety of purpose. Phytotoxicity study of the treated water was carried out and the study confirms non toxicity nature of the treated water. In‐silico screening was carried out to test the interaction efficacy of enzymes such as Laccase, Azo reductase, and peroxidase which are responsible for effective dye degradation with synthetic azo dyes. In silico studies showed better interactions between synthetic azo dyes and enzymes. This is the novel report on showing the effective degradation of synthetic azo yellow dye by using C. freundii isolated from tannery effluent.

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.

A two-photon mitochondria-targeting azo reductase probe for imaging in tumor cells and mice

Herein, a two-photon mitochondria-targeting fluorescent probe (TPNP) which can be activated by the azo reductase (AzoR) has been developed for imaging the hypoxic microenvironment of tumors in vivo. The probe TPNP showed a low detection limit (LOD = 1.85 eq), high selectivity, and considerable steadiness in different pH conditions (3.0–11.0). In living cells, it could co-locate with the commercial dye in the mitochondria (co-localization coefficient: 0.92) and indicate hypoxia in both one-photon and two-photon fluorescence imaging. Its AzoR-activation mechanism was further verified by the docking simulation as well as the experiments of adding AzoR or electron transporter inhibitors. Moreover, the in vivo real-time fluorescence imaging proved that TPNP could monitor hypoxia and the tumor progression in mouse tumor models. Overall, this designed probe exhibited high sensitivity, synthetic ease, and excellent fluorescent properties, which might therefore enlighten new ideas for the diagnosis of tumors in future studies.

Mechanistic insights on enzyme mediated-metabolite cascade during decolourization of Reactive Blue 13 using novel microbial consortium

Understanding the role of oxido-reductase enzymes followed by deciphering the functional genes and their corresponding proteins are crucial for the speculation of molecular mechanism for azo dye degradation. In the present study, decolourization efficiency of developed microbial consortium was tested using 100 mgL−1 reactive blue 13 (RB13) and the results showed ∼92.67% decolourization of RB13 at 48 h of incubation. The fourier-transform infrared spectroscopy (FTIR), high-performance liquid chromatography (HPLC) and gas chromatography–mass spectrometry (GC-MS) analysis were performed to identify the metabolites formed during RB13 degradation, followed by hypothesizing the metabolic pathway. The GC-MS analysis showed formation of 1,4-dihydronaphthalen-1-ol and 1,3,5-triazin-2-amine as the final degraded compounds after enzymatic breakdown of RB13 dye. The activity of different oxido-reductase enzymes was determined, and the results showed that NADH DCIP reductase and azo reductase had higher activity than other enzymes. It clearly indicated the degradation was initiated with the enzymatic cleavage of azo bond of RB13. Further, the functional genes were annotated against the database of clusters of orthologous groups (COGs) and kyoto encyclopedia of genes and genomes (KEGG). It provided valuable information about the role of crucial functional genes and their corresponding proteins correlated with dominant bacterial species in degradation of RB13. Hence, the present research is the first systematic study that correlated the formation of degradation compounds with the functional genes/enzymes and their corresponding bacterial species responsible for RB13 degradation.

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.

Bacterial enzymes for azo dye degradation: an insight

Azo dye has a massive marketplace in various industries, biological laboratories and clinics. A significant drawback is the excessive flush-off of azo dyes and their intermediate products are that they pollute the environment. Several environmental bacteria degrade azo dye under certain specific ecological conditions. Decolourisation by different bacteria occurs under aerobic, anoxic and anaerobic conditions. Bacterial genera such as Pseudomonas, Bacillus and Rhodococcus are used for the rapid decolourisation of azo dyes. Bacterial enzymes play an essential role in the degradation of azo dyes by cleavage of azo bonds (-N=N-). It is degraded by both oxidative and reductive enzymes including azo reductase, laccase, lignin peroxidase, NADH-DCIP reductase, malachite green reductase, polyphenol oxidase (tyrosinase), veratrol alcohol oxidase. This review describes the role and mechanism of these enzymes in the degradation of azo dyes.

Biodegradation and Detoxification of Azo Dyes by Halophilic/Halotolerant Microflora Isolated From the Salt Fields of Tibet Autonomous Region China.

This study aimed to decolorize azo dyes in high-salt industrial wastewater under high-salt and low oxygen conditions using extreme halophilic/halotolerant bacteria screened from the salt fields of Tibet, which consisted of Enterococcus, unclassified Enterobacteriaceae, Staphylococcus, Bacillus, and Kosakonia. Under the optimal conditions, 600 mg/l Congo red, Direct Black G (DBG), Amaranth, methyl red, and methyl orange could be completely decolorized in 24, 8, 8, 12, and 12 h, respectively. When the DBG concentration was 600 mg/l, NADH–DCIP, laccase, and azo reductase were confirmed to be the primary reductase and oxidase during the degradation process, and the degradation pathways were verified. The microflora could not only tolerate changes in salt concentrations of 0–80 g/l, but also displayed strong degradative ability. Under high-salt concentrations (≥ 60 g/l NaCl), NADH–DCIP reductase was primarily used to decolorize the azo dye. However, under low salt concentrations (≤ 40 g/l NaCl), azo reductase began to function, and manganese peroxidase and lignin peroxidase could cooperate to participate in DBG degradation. Additionally, the halophilic/halophilic microflora was shown to convert the toxic DBG dye to metabolites of low toxicity based on phytotoxicity analysis, and a new mechanism for the microflora to degrade DBG was proposed based on intermediates identified by liquid chromatography-mass spectrometry (LC–MS). This study revealed that the halophilic/halophilic microflora has effective ecological and industrial value for treating wastewater from the textile industry.

Biochemical characterization of a novel azo reductase named BVU5 from the bacterial flora DDMZ1: application for decolorization of azo dyes.

One of the main mechanisms of bacterial decolorization and degradation of azo dyes is the use of biological enzymes to catalyze the breaking of azo bonds. This paper shows the expression and properties of a novel azo reductase (hybrid-cluster NAD(P)-dependent oxidoreductase, accession no. A0A1S1BVU5, named BVU5) from the bacterial flora DDMZ1 for degradation of azo dyes. The molecular weight of BVU5 is about 40.1 kDa, and it contains the prosthetic group flavin mononucleotide (FMN). It has the decolorization ability of 80.1 ± 2.5% within 3 min for a dye concentration of 20 mg L−1, and 53.5 ± 1.8% even for a dye concentration of 200 mg L−1 after 30 min. The optimum temperature of enzyme BVU5 is 30 °C and the optimum pH is 6. It is insensitive to salt concentration up to a salinity level of 10%. Furthermore, enzyme BVU5 has good tolerance toward some metal ions (2 mM) such as Mn2+, Ca2+, Mg2+ and Cu2+ and some organic solvents (20%) such as DMSO, methanol, isopentyl, ethylene glycol and N-hexane. However, the enzyme BVU5 has a low tolerance to high concentrations of denaturants. In particular, it is sensitive to the denaturants guanidine hydrochloride (GdmCl) (2 M) and urea (2 M). Analysis of the dye substrate specificity shows that enzyme BVU5 decolorizes most azo dyes, which is indicating that the enzyme is not strictly substrate specific, it is a functional enzyme for breaking the azo structure. Liquid chromatography/time-of-flight/mass spectrometry (LC-TOF-MS) revealed after the action of enzyme BVU5 that some intermediate products with relatively large molecular weights were produced; this illustrates a symmetric or an asymmetric rapid cleavage of the azo bonds by this enzyme. The potential degradation pathways and the enzyme-catalyzed degradation mechanism are deduced in the end of this paper. The results give insight into the potential of a rapid bio-pretreatment by enzyme BVU5 for processing azo dye wastewater.

Construction and implementation of floating wetpark as effective constructed wetland for industrial textile wastewater treatment

Fimbristylis dichotoma, Ipomoea aquatica, Pluchea tomentosa and their co-plantation (consortium FIP) autonomously degrade Orange 3R. Consortium FIP showed 84% removal of Orange 3R within 48 h, which is a higher dye elimination rate than individual plant systems. Oxidoreductase enzymes like tyrosinase (76%), varatryal alcohol oxidase (85%), lignin peroxidase (150%), riboflavin reductase (151%), laccase (171%), NADH-DCIP reductase (11%) and azo reductase (241%) were expressed in consortia FIP during Orange 3R degradation. UV–vis spectroscopy, enzyme activities, HPTLC, FTIR and GC-MS confirmed mineralization of Orange 3R into its metabolites. Microscopic investigation of root tissue revealed the harsh effect of dye on root tissues. Toxicity assessment on the HepG2 cell line demonstrated the toxic nature of Orange 3R, which gets reduced after phyto-treatment with consortia FIP. Floating wetpark of consortia FIP was found more efficient for the treatment of industrial textile waste and accomplished 87%, 86%, 75%, 49% and 46% removal of COD, BOD, color, TSS and TDS of effluent.

Synthesis of magnetic hydrochar from Fenton sludge and sewage sludge for enhanced anaerobic decolorization of azo dye AO7

A novel magnetic hydrochar synthesized from Fenton sludge (FS) and sewage sludge (SS) was employed in the anaerobic decolorization of acid orange 7 (AO7). The stable presence of Fe3O4 in magnetic hydrochar was confirmed by physicochemical characterization. The degradation efficiency of AO7 in the anaerobic system with the addition of hydrochar prepared in an optimal proportion (SS:FS=1:3, named as HC-1:3) could reach 98.55%, which was 1.91 times higher than the control system. Particularly, superior electrical conductivity, electron transport system activity and azo reductase activity of the sludge in anaerobic system with HC-1:3 were achieved. The redox of Fe(Ⅲ)/Fe(Ⅱ) in anaerobic system was realized by dissimilatory iron-reducing bacteria enriched with HC-1:3. According to the six-cycle batch experiments and 120-day continuous-flow UASB experiments, the addition of HC-1:3 into the anaerobic system facilitated the diversity of microbiological community and increased the ecological stability of anaerobic system. The possible electron transfer mechanism involving in the magnetic hydrochar-based anaerobic system for AO7 removal was speculated preliminarily. The as-prepared magnetic hydrochar not only showed a promising future in anaerobic system for recalcitrant contaminants degradation, but also provided a new approach for the resource utilization of FS and SS.

Biodegradation of Brown 706 Dye by Bacterial Strain Pseudomonas aeruginosa

Dyes are the most challenging pollutants for the aquatic environment that are not only toxic, but also interfering photosynthesis as light penetration into deep water is changed. A number of methods are used for the water reclamation, however, among them biological methods are preferably used due to their compatibility with nature. In the present research, 15 different bacterial strains were used to decolorize Brown 706 dye. Among the bacterial strains, Pseudomonas aeruginosa showed maximum decolorization activity; hence in the subsequent experiments Pseudomonas aeruginosa was used. First the decolorization activities were carried out under different physicochemical conditions to obtain the optimum decolorization benefits of the selected microorganism. The optimum conditions established were 37°C, pH of 7 and operation cycle time 72 h. In the subsequent experiment all optimum conditions were combined in a single experiment where 73.91% of decolorization efficiency was achieved. For the evaluation of metabolites formed after decolorization/degradation the aliquots containing bacteria were homogenized, filtered and then subjected to extraction. The extracted metabolites were then subjected to the silica gel column isolation. UV–Vis, FTIR, and NMR techniques were used to elucidate structures of the metabolites. Out of the collected metabolites only P-xylene was identified, which has been formed by cleavage of azo linkage by azo reductase enzyme of bacteria following the deamination and methylation of nitro substituted benzene ring.

Combining acetone-butanol-ethanol production and methyl orange decolorization in wastewater by fermentation with solid food waste as substrate

Simultaneously producing acetone-butanol-ethanol (ABE) and decolorizing Methyl orange (MO) by fermentation with solid food waste (SFW) was studied. During the batch fermentation with MO concentration from 100 mgL−1 to 1000 mgL−1, carbon source utilization efficiency was above 93% with yields of acetone, butanol and ethanol being 0.13 gg-1, 0.23 gg-1 and 0.04 gg-1, respectively. During the repeated fermentation with the bagasse as the cell immobilization material, carbon source utilization efficiencies were all above 90% in the first 5 cycles fermentation. The maximum butanol and ABE concentration were 12.8 gL-1 and 22.1 gL-1, respectively. The solvent yield of acetone, butanol and ethanol were around 0.13 gg-1, 0.25 gg-1 and 0.06 gg-1, respectively. The MO decolorization had no negative effect on the ABE production and the decolorization efficiencies were all above 99% during batch fermentation or repeated fermentation with electrons transferred from intracellular to extracellular by mediator with the help of azo reductase to decolorize MO.

Eco-friendly bioprocess using agar plug immobilized Penicillium crustosum PWWS-6 biomass for treatment of wastewater contaminated with toxic Congo red dye for use in agriculture

Recycling and valorization of noxious dye contaminated industrial wastewater minimize prevailing agricultural water dearth. In this context, the current manuscript investigates and fabricates a rapid, ecofriendly, and cost effective treatment system using a potent dye degrading microbial agent. An agar plug immobilized Penicillium crustosum PWWS-6, a filamentous fungus isolated from veterinary clinical waste dump, for the first time reported to decolorize (81.86 ± 0.03 %) hazardous diazo dye, (Congo red, 100 ppm) in 24 h. Lignin peroxidase, azo reductase, veratryl alcohol oxidase, and NADH-DCIP reductase enzymes were induced significantly during mycoremediation bioprocess which is affected by incubation time, temperature, pH, inoculum volume, and dye concentration of aqueous medium. The one-factor-at-a-time (OFAT) and response surface methodology (RSM) were adopted to optimize these cultural parameters and generated optimized conditions (incubation time, 16 h; temperature, 27 °C; and inoculum volume, 6 fungal discs), for maximum degradation (99.85 %) of 100 ppm CR at pH 7.0. Further, using OFAT approach, P. crustosum also showed 72.14 % degradation of CR at much higher concentration of 1000 ppm in 72 h. UV–vis spectra verified complete disappearance of dye from colored broth. Fourier Transform Infrared (FTIR) spectroscopy along with Field Emission Scanning Electron Microscopy (FESEM) subsequently confirmed (–N=N–) bond destabilization following degradation. Furthermore, the non-toxic degraded metabolites showed improved percent germination, radical and plumule length of Cicer arietinum and Hordeum vulgare seeds compared with untreated CR contaminated wastewater.

Microbial degradation of azo dyes by textile effluent adapted, Enterobacter hormaechei under microaerophilic condition

Landmark and sustainable eco-friendly dye treatment processes are highly desirous to ameliorate their effect on the environment. The present study investigated the azo dye degradation efficiency of adapted Enterobacter hormaechei SKB 16 from textile effluent polluted soil in optimized culture conditions. The adapted bacteria strain was identified by standard microbiological and molecular techniques. E. hormaechei was tested individually for the decolourizing of Reactive Yellow 145 (RY 145) and Reactive Red F3B (RR 180) dyes under optimized conditions of pH, temperature and dye concentration on decolourization were studied. The adapted bacteria strain exhibited maximum decolourization (98 %) of Reactive yellow 145 and Reactive red 180 in 100 ppm concentration at pH 7, temperature 37 °C after 98 h of incubation. The enzyme analyses revealed that azo reductase and laccase played major roles in the cleavage of the azo bond and desulfonation respectively of both dyes during degradation. The metabolites were further characterized by Fourier Transform Infrared Spectroscopy (FT-IR), High-Performance Liquid Chromatography (HPLC), and Gas Chromatography-Mass Spectrometry (GCMS). Thereafter, degradation was deduced based on changes of the functional group, variation in retention times and mass/charge ratio and molecular weight. This study elucidated the promising potentials of adapted SKB 16 strain in the eco-friendly removal of textile azo dyes. In addition, repeatability and sustainability are enhanced due to effective management of time which would have been spent on rigorous and extensive screening process.