Allura Red Research Articles

A brief review on recent high-performance platforms for electrochemical sensing of azo dye Allura Red (E129): food safety and pharmaceutical applications

Artificial dyes are increasingly widespread, especially in foodstuffs and pharmaceutical products as an industry marketing strategy to attract consumers. Consumption of the synthetic azo dye Allura Red (AR) has potential risks to human health and can cause several adverse health effects such as genotoxicity, carcinogenicity, attention deficit hyperactivity disorder, allergic and asthmatic diseases in children. The limited number of electrochemical sensing platforms for AR successfully applied for safety assessment of foods, beverages, and drug formulations testifies that the food and pharmaceutical matrices present significant analytical challenges and there is still a need to amplify the analytical performances of these systems. The authors intensively reviewed recent papers (mainly since 2019) emphasizing electrode engineering, strategies for electrochemical signal amplification, and analytical applications in food and drug control. A critical discussion on the latest interesting innovations and perspectives of the most promising electrochemical tools for AR electro­analysis is presented. The challenges and limitations in the design of electrochemical sensors for AR analysis are also discussed with a view to providing new directions for future research and development of sustainable sensing devices, paving the way for advancements in this field.

Illumination of biosorption of Allura Red on Rhus coriaria L. (sumac) plant: equilibrium, kinetic, thermodynamic, and density functional theory–based analyses

Illumination of biosorption of Allura Red on Rhus coriaria L. (sumac) plant: equilibrium, kinetic, thermodynamic, and density functional theory–based analyses

Evaluating the potential of stabilised betacyanins from fermented red dragon fruit (Hylocereus polyrhizus) drink: Sustainable colouration and antioxidant enhancement of stirred yoghurt

Nowadays, the wide use of synthetic food colourant, Allura Red has elevated negative perceptions among consumers. As food industry players strive towards satisfying consumers’ demands, exploring new sustainable natural colourants with multifunctional properties (like betacyanins) has emerged as a novel concept. This study aimed to evaluate the potential of a functional fruit drink, Improved-FRDFD-dH2O [contained betacyanins of fermented red dragon fruit drink stabilised by 0.4 % xanthan gum and 0.5 % carboxymethyl cellulose hydrocolloid mixture solution and 0.2 % citric acid] to serve as a sustainable and stable functional liquid colourant in yoghurt. The 8-week refrigerated storage study revealed the betacyanins content in the yoghurt added with E162 experienced the lowest stabilities in terms of betacyanins content (>30 % degradation), viscosity (>15 % reduction), pH, LAB viability and antioxidant activity. In contrast, the Improved-FRDFD-dH2O was able to maintain the yoghurt's viscosity, syneresis, pH and LAB viability without any significant change. The betacyanins of Improved-FRDFD-dH2O had excellent stability in the yoghurt (<5 % degradation), showing only a minimal difference (ΔE<1.5) in total colour changes and being able to retain >86 % of the initial antioxidant activity by the end of storage. These findings support the Improved-FRDFD-dH2O as a sustainable functional colourant with antioxidant properties for producing beneficial yoghurt.

Changes Caused in Liver Parameters in Swiss Albino Mice by Food Colourant (Allura Red) and Preservative (Sodium Benzoate)

Food additives are chemicals that are purposefully added to food to improve its quality. The most widely used food additives in the food industry, pharmacy and cosmetics field are Allura red (a food colour) and sodium benzoate (a food preservative), both of which are employed for various functions such as colouring and preserving. In this study, liver parameters were examined in Swiss male mice after 30, 45 and 60 days of treatment with synthetic Allura red colour and sodium benzoate preservative. Seventy-two male Swiss albino mice (25-30 gm) were divided into four groups with 6 mice per group for each autopsy interval. The first group was control-treated (water), the second group was Allura red treated (172.2 mg/kg body weight), the third group was sodium benzoate treated group (123 mg/kg body weight) and the fourth group was Allura red + sodiumbenzoate treated (86.1 mg/kg + 61.5 mg/kg body weight) for 30, 45 and 60 days. Mice were sacrificed after the 31st, 46th and 61st day and blood samples were taken for serological tests to study SGPT, SGOT, ALP, total protein and serum albumin. The liver was also removed for histological studies of control, sodium benzoate, Allura red and combination groups. The data obtained reveals a noticeable significant increase in aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total protein and albumin in treated mice as compared to control groups. The microscopic study of the liver showed the effect of all dose treatments on the structure of liver cells. It caused fatty degeneration of hepatocytes, congestion of the central vein and necrosis. In conclusion, it was clear that the administration of Allura red, sodium benzoate, and combination dose (AR+SB) in mice caused changes in liver parameters.

Statistical physics insights in the Allura Red adsorption on chitosan: Effects of adsorbents features, and experimental conditions

This study presents an innovative approach using statistical physical modeling to analyze the effect of experimental conditions and adsorbent features on removing Allura Red dye by chitosan. Five models based on statistical physics were employed to understand the removal mechanism under different experimental conditions, including variations in pH, particle sizes, chitosan deacetylation degrees, and temperatures. The best models were determined through rigorous statistical analysis and a detailed evaluation of the estimated parameters, considering their physical meaning and standard deviations. The models demonstrated high predictive capacity (coefficients of determination greater than 0.9 and low mean squared error values < 110). Analysis of the effects of experimental conditions revealed that a pH of 6.6, particles smaller than 0.10 mm, a high degree of deacetylation (84 %), and temperatures of 298 K and 308 K present the greatest potential for dye removal by chitosan. Furthermore, it was observed that, in general, the dye removal mechanism involves the formation of a double layer with two adsorption energies under the best experimental conditions, and the process occurs predominantly by physical forces (adsorption energy below 35 kJ/mol). This work contributes significantly to understanding how experimental conditions influence the mechanism of dye adsorption by chitosan, offering valuable and new insights for future applications and optimizations in the treatment of effluent-containing dyes.

Rapid ion pair-based surfactant-assisted liquid-liquid microextraction with solidification of floating organic drop for simultaneous spectrophotometric determination of selected anionic dyes in food samples

Ion pair-based surfactant-assisted liquid-liquid microextraction with solidification of floating organic drops has been developed to extract Allura red (AR), tartrazine (TAR), and fast green (FG) prior to spectrophotometric determination. Cetyltrimethylammonium bromide (CTAB) was employed as ion-pairing agent to enhance the hydrophobic behavior of anionic dyes. 1-undecanol and ethanol were used as the extraction and dispersion solvents, respectively. The dyes were quantitatively extracted in the presence of KCl (0.15 mol L−1) at pH 4.0. The method exhibits wide linearity (15.0–1500.0 μg L−1 for AR, 35.0–2000.0 μg L−1 for TAR, and 3.0–1200.0 μg L−1 for FG) with preconcentration factors of 19.6, 20.1, and 19.9, respectively. The detection limit was 3.7. 9.5, and 0.83 μg L−1 for AR, TAR, and FG, respectively. The relative standard deviation did not exceed 2.1 %. The procedure was applied for the determination of these dyes in food samples.

Impact of dye treatment as management strategy on available light may favour a highly invasive alien aquatic plant

Attempts to control massive proliferations of invasive alien aquatic plants (IAAP) are often ineffective. A renewed interest for dye treatment is emerging aiming to control the submerged macrophyte Myriophyllum heterophyllum in France. We aimed to understand the effects of dye on this plant knowing about its adaptation to low light. In a 2 × 2 factorial design experiment, we assessed the effect of a dye mixture based on Brilliant Blue (E133) and Allura Red (E129) at high and low light intensities on light quantity and quality and how this might affect the plant’s performance by measuring morphological and physiological traits. A multivariate analysis identified three groups – high light (HL) plants, plants in high light with dye (HLD) or low light (LL), and low light with dye (LLD) plants. HL plants performed well but showed stress signs, with a reduced main shoot length, a higher leaf dry matter content (LDMC) and a lower nitrogen content (N), not observed in plants grown in HLD or LL. HLD and LL plants exhibited a comparable total length growth rate to those in HL, but had lower LDMC and higher N contents. LLD plants performed poorly with the lowest growth and signs of physiological stress. Dye-induced changes in light quality only marginally affected the absorbance range of chlorophyll b, which apparently did not affect photosynthesis. Commercially available dyes currently used to control nuisance aquatic plants thus seem to have little or no effect on submerged macrophytes. The presence of dye may exacerbate negative effects of very low light intensities on the plant’s growth. However, these very low intensities would only be reached during high-water levels or in winter, periods where the dye would rapidly be diluted. During summer, however, the application of dye may protect the plant from damaging light intensities and thus not be a good management strategy to control low light adapted invasive submerged macrophytes.

Photogalvanics of the platinum working electrode in the Allura Red d-Galactose-didecyl dimethyl ammonium chloride-NaOH electrolyte

Photo-galvanic cell is a photochemical cell device whose performance depends on the combination of several factors like dye and reductant concentrations, dye stability, pH, light intensity, electrodes used, Pt electrode area, diffusion length, etc. An entirely new and unexplored combination of platinum with Allura Red photosensitizer, d-Galactose reductant, and didecyl dimethyl ammonium chloride (DDAC) surfactant has been explored in present research to upgrade the electrical performance of these cells. Allura Red is a water-soluble azo dye which shows good absorbance in region 501–507 nm. The Allura Red and DDAC is an anionic and cationic species, respectively, and therefore, the opposite charges of dye and surfactant molecules are expected to form a stable dye-surfactant complex enhancing the dye solubility and stability. d-Galactose has been used for its good reducing properties. This unexplored combination of Pt-Allura Red-d-Galactose-DDAC with these characteristics has encouraged further enhancement of the electrical output of the photogalvanic cells. The electrical output, stability, and spectral property of the photogalvanic cells have been studied in the present work. The observed power, current, potential, efficiency, and storage capacity (as half time) are of the order of 443.8 µW, 2400 µA, 721 mV, 11.61%, and 28 min, respectively. These observed results are higher than some reported data. In the spectral study, nearly the same band intensity of the pre-illuminated and post-illuminated electrolyte solution shows quite good photo-stability of the Allura Red dye in electrolyte form. This new combination of platinum-electrolyte still has the scope to achieve the enhanced cell performance of photogalvanic cells for future development.

Dispersive liquid–liquid microextraction based on solidification of floating organic droplet and spectrophotometric determination of Allura red using green deep eutectic solvent as disperser solvent: Green profile assessment

In this study, dispersive liquid–liquid microextraction based on the solidification of a floating organic drop (DLLME-SFO) was investigated for allura red (AR) preconcentration from water and food samples using green deep eutectic solvent (DES) as a disperser solvent. Compared to the conventional disperser solvents, the DES, which consists of choline chloride and citric acid (1:1), exhibited a higher AR recovery with simple preparation, being eco-friendly and low expense. The proposed protocol showed relatively high recovery values of AR in real samples (97.0–99.5 %). The optimum conditions for AR extraction were pH=3.0, 250.0 µl DES volume, CTAB (4.0 × 10−4 mol L−1), KCl (0.10 mol L−1), 100 µl of 1-dodecanol, and centrifugation: 4.0 min. at 4000 rpm. The validated method showed linearity from 100 to 5000 µg L−1 with 14.4 enrichment factor for 10.0 mL sample. The limit of detection was 29.9 µg L−1 and the relative standard deviation was 3.7 % for five replicate analysis of 300.0 µg L−1. The AR content in studied food samples was determined by spectrophotometry and high-performance liquid chromatography (HPLC). Our proposed method is simple and fast, as the whole process for AR extraction requires only 9.0 min. The analytical eco-scale AGREEnness calculator (AGREE), the blue applicability grade index (BAGI), and the ChlorTox scale were evaluated.

Allura Red AC is a xenobiotic. Is it also a carcinogen?

Merriam-Webster and Oxford define a xenobiotic as any substance foreign to living systems. Allura Red AC (a.k.a., E129; FD&C Red No. 40), a synthetic food dye extensively used in manufacturing ultra-processed foods and therefore highly prevalent in our food supply, falls under this category. The surge in synthetic food dye consumption during the 70s and 80s was followed by an epidemic of metabolic diseases and the emergence of early-onset colorectal cancer in the 1990s. This temporal association raises significant concerns, particularly given the widespread inclusion of synthetic food dyes in ultra-processed products, notably those marketed toward children. Given its interactions with key contributors to colorectal carcinogenesis such as inflammatory mediators, the microbiome, and DNA damage, there is growing interest in understanding Allura Red AC's potential impact on colon health as a putative carcinogen. This review discusses the history of Allura Red AC, current research on its effects on the colon and rectum, potential mechanisms underlying its impact on colon health, and provides future considerations. Indeed, although no governing agencies classify Allura Red AC as a carcinogen, its interaction with key guardians of carcinogenesis makes it suspect and worthy of further molecular investigation. The goal of this review is to inspire research into the impact of synthetic food dyes on colon health.

Chitosan-azo dye bioplastics that are reversibly resoluble and recoverable under visible light irradiation.

Biopolymer composite materials were prepared by combining bio-sourced cationic water-soluble chitosan with bi-functional water-soluble anionic azo food dyes amaranth (AMA) or allura red (ALR) as ionic cross-linkers, mixing well in water, and then slow-drying in air. The electrostatically-assembled ionically-paired films showed good long-term stability to dissolution, with no re-solubility in water, and competitive mechanical properties as plastic materials. However, upon exposure of the bioplastics to low power light at sunlight wavelengths and intensities stirring in water, the stable materials photo-disassembled back to their water-soluble and low-toxicity (edible) constituent components, via structural photo-isomerization of the azo ionic crosslinkers. XRD, UV-vis, and IR spectroscopy confirmed that these assemblies are reversibly recoverable and so can in principle represent fully recyclable, environmentally degradable materials triggered by exposure to sunlight and water after use, with full recovery of starting components ready for re-use. A density functional theory treatment of the amaranth azo dye identified a tautomeric equilibrium favouring the hydrazone form and rationalized geometrical isomerization as a mechanism for photo-disassembly. The proof-of-principle suitability of films of these biomaterial composites as food industry packaging was assessed via measurement of mechanical, water and vapour barrier properties, and stability to solvent tests. Tensile strength of the composite materials was found to be 25-30 MPa, with elongation at break 3-5%, in a range acceptable as competitive for some applications to replace oil-based permanently insoluble non-recyclable artificial plastics, as fully recyclable, recoverable, and reusable low-toxicity green biomaterials in natural environmental conditions.

Trachyspermum ammi Mediated Green Synthesis of NiO Nanoparticles: Dual Role in Photocatalytic Degradation and Anti-bacterial Efficacy

ABSTRACT Nowadays, there is a lot of discussion in science about green chemistry of nanomaterials, from synthesis to various environmental applications. Herein, we describe a simple, environmentally friendly, economically feasible, free of additives, phytosynthetic methodology for the synthesis of cubic phased NiO nanoparticles using the seed extract of Trachyspermum ammi (also known as Ajwain), which acts as a capping and reducing agent that helps in reducing nickel sulphate (NiSO4.6 H2O) to NiO via co-precipitation method. Various spectroscopic techniques were involved in investigating the phase, morphology, functional groups, crystallinity, size, elemental confirmation, optical property with band gap of the green synthesised NiO Nanoparticles. Moreover, green NiO nanoparticles displayed excellent degradation efficiency for the reduction of industrial pollutant, Allura red and possible mechanism were also suggested. Nearly 91.73% of the Allura red was degraded using the synthesised NiO nanoparticles within 90 min when exposed to UV-light. Moreover, the used nanoparticles were renewed and again their photocatalytic performance was assessed for three consecutive cycles. In addition to photocatalytic activity, the green NiO nanoparticles revealed the considerable anti-bacterial activity against Staphylococcus aureus and Escherichia coli and the dead percentage was calculated as 92.56% and 94.33% respectively. In this work, green NiO nanoparticles was involved as a dual functioning agent for effective photocatalytic and anti-bacterial studies.

Simultaneous determination of 11 water-soluble synthetic colorants in foods consumed in Chile by high-performance liquid chromatography with diode Array detection

In Chile, limited information is available on colorants in commonly consumed foods among vulnerable age groups. We developed and validated a rapid HPLC-DAD method to simultaneously evaluate 11 synthetic colorants in candies, beverages, ice cream, and cereals. The method exhibited excellent analytical performance for all 11 colorants with LOD (0.44 - 1.55 mgL-1), LOQ v(1.32 - 4.70 mgL-1), precision (4.0 and 7.3% RSD), and recovery (80 - 105%) in fortified matrices (10-50-100 mgL-1). The highest detection frequencies were as follows: cereals > candies > beverages > ice cream. Sunset Yellow was the most prevalent colorant in all food matrices, followed by Allura Red and Azorubine. Positive samples contained between 1 and 5 synthetic colorants. With the exception of cereals, the colorant concentrations in the remaining matrices exceeded the Codex Alimentarius regulations and the values reported in other studies worldwide, indicating the Chilean population is at risk.

Simultaneous deep eutectic solvent based microextraction of Allura Red and Brilliant Blue in violet-purple foodstuffs

A vortex-assisted simultaneous deep eutectic solvent based microextraction and spectrophotometric determination method was established for the first time to simultaneously monitor Allura Red and Brilliant Blue in violet-purple foodstuffs. A ternary deep eutectic solvent was created using benzoic acid, tetra butyl ammonium bromide and benzyl alcohol at 1:1:3 mol ratio. The method variables including pH, volume of deep eutectic solvent and vortex time were optimized by Box-Behnken design of response surface methodology. The limits of detection were determined to be 25 and 9 µg/L for Allura Red and Brilliant Blue, respectively. The linear dynamic ranges of the method were obtained between 82 and 12,000 and 29–4200 µg/L for Allura Red and Brilliant Blue, respectively. Preconcentration factor of the method was obtained as 15. Greenness evaluation was performed using Analytical GREEnness tools. Extraction mechanisms affecting quantitative simultaneous microextraction of the target dyes were explained. The developed method was validated with high-performance liquid chromatography analysis. Analyte addition-recovery tests were also applied to confectionery samples to validate the proposed method and determine the dye contents of foodstuffs. Allura Red and Brilliant Blue contents of violet-purple confectioneries were determined between 3.85 and 74.59 and 4.02–26.11 µg/g concentrations with 94.5–98.6 % and 93.0–101.0 % recoveries, respectively.

A sensitive immunochromatography assay based on quantum dot nanobeads for determination of acid orange II

Acid orange II (AO2) is a synthetic azo dye commonly used illegally in food production and processing. The development of a fast and effective analytical method is crucial for controlling the unlawful use of AO2 in food, especially in seasoning. Here, an antibody specific for AO2 was generated, with 1.01 ng/mL IC50 and minimal cross-reactivity with other analogs (<1 %, including sunset yellow, tartrazine, para red, chromotrope FB, Sudan Ⅰ, rhodamine B, 2-naphthol, amaranth, ponceau 4 R and allura red AC). Furthermore, a highly sensitive quantum dot beads-based fluorescence immunochromatography assay (QBs-FICA) was established for AO2 determination. The assay had a working range of 23–287 μg/kg with a 11 μg/kg limit of detection (LOD) in chili powder and hotpot seasoning samples. Using spiked samples, the results of QBs-FICA were found to be consistent with those of liquid chromatography-tandem mass spectrometry. This indicates that QBs-FICA is a reliable, rapid, and portable tool for testing of AO2.

Investigation on Deterioration of Allura Red Dye Under Visible Radiation by TiO2 Based Nanocomposite

This current study is to prepare and characterise a Cu-doped Titanium dioxide (TiO2) and Cu-doped Titanium dioxide with Tungsten (WO3) nanocomposite and its application for the deterioration of Allura Red (AR) Dye. After preparing the Cu- doped Titanium dioxide (TiO2) with WO3 and Cu-doped Titanium dioxide (TiO2) nanocomposites using wet chemical Sol-gel method over various soaking times, the XRD, BET, and Ultraviolet-visible spectroscopy were used to determine the properties of the synthesized Copper-doped Titanium dioxide and Copper-doped Titanium dioxide with Tungsten nanocomposites. When Tungsten was added to Cu-doped Titanium dioxide (TiO2), the X-Ray Diffraction showed that all of the composites' peaks shifted slightly in the direction of a higher diffraction angle. To evaluate surface area BET curve was used. The Copper-doped Titanium dioxide (TiO2)/Tungsten (WO3) nanocomposite gives a significant photocatalytic activity because of its increased surface area. High surface area (BET) and anatase phase development (XRD), which are the main contributors to effective photocatalytic activity, were identified by the work's results. XRD revealed that the synthesised materials were in the anatase phase. Compared to CDT, CTW-15, CTW-30, and CTW-60 nanocomposites, the surface area (BET) of the CTW-45 nanocomposite is larger at 128.79 m2/g. By examining the kinetics of reaction parameters such as dopant concentrations, pH, catalyst dose, and dye concentration, the catalysts were able to determine the ideal conditions for improved photocatalytic degradation of Allura Red dye. At pH 4, dye concentrations of 5 mg/L, and CTW-45 concentrations of 100 mg/L, the percentage of AR dye degradation was greater. The findings of this study can be used to develop efficient nanocomposites that remove Allura Red dye from wastewater.

Food Safety and Health Concerns of Synthetic Food Colors: An Update.

The toxicity of food additives is widely studied and concerns many consumers worldwide. Synthetic food colors are often considered an unnecessary risk to consumer health. Since the European Food Safety Authority's (EFSA) re-evaluation between 2009 and 2014, the body of scientific literature on food colors has grown, and new evaluations are being published by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Therefore, this narrative review aims to review the toxicological data that have become available since 2014. The reviewed colors are Quinoline Yellow, Sunset Yellow, Azorubine, Amaranth, Ponceau 4R, Erythrosine, Allura Red, Patent Blue, Indigo Carmine, Brilliant Blue FCF, Green S, Brilliant Black, Brown HT, and Lithol Rubine BK. Tartrazine was not included in this paper; the overwhelming amount of recent data on Tartrazine toxicity requires more space than this review can provide. The issues regarding the toxicity of synthetic food colors and real population exposures are being regularly examined and reviewed by relevant authorities, such as the EFSA and JECFA. The current ADI limits set by the authorities are mostly in agreement, and they seem safe. However, the EFSA and JECFA assessments of some of the colors are more than a decade old, and new evidence will soon be required.

Low-Cost Solar Still System with Concave Condensing Cover for the Distillation of Synthetic Water Polluted with Allura Red Dye

Low-Cost Solar Still System with Concave Condensing Cover for the Distillation of Synthetic Water Polluted with Allura Red Dye

Fast on-Site and Highly Sensitive Determination of Allura Red Using a Portable Electrochemical Platform Based on Poly(pyrrole) and Bimetallic Nanocomposites Anchored on Reduced Graphene Oxide

Allura red (AR) is classified as an azo dye and is often used as a beverage and food additive. Nevertheless, the need for dose management of Allura red becomes especially important owing to the potential damage caused by the azo structure to the human body and the environment. In order to combat these problems, a novel portable electrochemical platform using a screen-printed carbon electrode (SPCE) that has been modified with poly(pyrrole) and Co-Ni bimetallic nanocomposites anchored on reduced graphene oxide (Co-Ni@rGO) was developed. The purpose of this platform is to enable rapid on-site and very sensitive determination of Allura red from carbonated energy beverages and water samples. Under ideal experimental conditions, the proposed platform’s response exhibits a notable linear relationship with the concentration of Allura red within the range of 0.0001–10 μM, having a very low limit of detection (LOD) of 0.03 nM and a high sensitivity of 24.62 μA μM−1 cm−2. Furthermore, the PPy/Co-Ni@rGO/SPCE platform exhibited favorable characteristics in terms of reproducibility, repeatability, stability, and selectivity for the quantification of Allura red. Consequently, the developed platform was capable of practically and effectively determining the Allura red dye content from various real samples, showing satisfactory recovery rates.

Selective pressure leads to an improved synthetic consortium fit for dye degradation

Microorganisms have great potential for bioremediation as they have powerful enzymes and machineries that can transform xenobiotics. The use of a microbial consortium provides more advantages in application point of view than pure cultures due to cross-feeding, adaptations, functional redundancies, and positive interactions among the organisms. In this study, we screened about 107 isolates for their ability to degrade dyes in aerobic conditions and without additional carbon source. From our screening results, we finally limited our synthetic consortium to Gordonia and Rhodococcus isolates. The synthetic consortium was trained and optimized for azo dye degradation using sequential treatment of small aromatic compounds such as phenols that act as selective pressure agents. After four rounds of optimization with different aims for each round, the consortium was able to decolorize and degrade various dyes after 48 h (80%–100% for brilliant black bn, methyl orange, and chromotrop 2b; 50–70% for orange II and reactive orange 16; 15–30% for chlorazol black e, reactive red 120, and allura red ac). Through rational approaches, we can show that treatment with phenolic compounds at micromolar dosages can significantly improve the degradation of bulky dyes and increase its substrate scope. Moreover, our selective pressure approach led to the production of various dye-degrading enzymes as azoreductase, laccase-like, and peroxidase-like activities were detected from the phenol-treated consortium. Evidence of degradation was also shown as metabolites arising from the degradation of methyl red and brilliant black bn were detected using HPLC and LC-MS analysis. Therefore, this study establishes the importance of rational and systematic screening and optimization of a consortium. Not only can this approach be applied to dye degradation, but this study also offers insights into how we can fully maximize microbial consortium activity for other applications, especially in biodegradation and biotransformation.