Abstract:In the present study, biotransformation of Remazol Orange 3R (RO3R) was studied using well-known bacterial isolate Pseudomonas aeruginosa strain BCH. The dye was decolorized up to 98 % within 15 min. The induction in the level of various oxidoreductive enzymes viz. laccase, tyrosinase, veratryl alcohol oxidase and DCIP reductase were observed in the cells obtained after decolorization of RO3R, which supports their role in decolorization. The metabolites of RO3R obtained after biodegradation were identified and… Show more
“…The analysis showed the occurrence of new peaks with disappearance of the control peaks in malachite green and methylene blue confirming effective decolorization by laccase whereas in case of congo red decolorization, appearance of new peaks along with detainment of control peak indicate slow and incomplete decolorization. The dye decolorization was supported with the help of HPLC analysis by various researchers earlier [23], [51], [52]. However, further study on identification of metabolites formed during dye decolorization is necessary.Figure 6HPLC analysis of Malachite green control (A), metabolites from Malachite green decolorization (B), Methylene blue control (C), metabolites from Methylene blue decolorization (D), Congo red control (E), metabolites from Congo red decolorization (F).…”
In this study we report the purification of laccase produced by Trichoderma harzianum strain HZN10 (using wheat bran under solid state fermentation) and its application in decolorization of synthetic dyes. Extracellular laccase was purified to homogeneity by DEAE-Sepharose and Sephadex G-100 chromatography with specific activity of 162.5 U/mg and 25-fold purification. Purified laccase was immobilized in various entrapments like calcium alginate, copper alginate, calcium alginate–chitosan beads and sol–gel matrix. Optimization results revealed that the laccase immobilized in sol–gel was optimally active in wide pH range (4.0–7.0) and thermo-stable (50–70 °C) than free enzyme which was optimum at 50 °C and pH 6.0. Kinetic analysis showed Km of 0.5 mM and 2.0 mM and Vmax of 285 U/mg and 500 U/mg by free laccase and sol–gel immobilized laccase respectively with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) [ABTS] substrate. Free and immobilized laccase was employed for decolorization of three different synthetic dyes (malachite green, methylene blue and congo red). High performance liquid chromatography (HPLC) analysis results revealed that approximately 100% of malachite green, 90% of methylene blue and 60% of congo red dyes at initial concentration of 200 mg/L were decolorized within 16, 18 and 20 h, respectively by laccase immobilized in sol–gel matrix in the presence of 1-hydroxybenzotriazole (HBT) mediator. During the decolorization all three synthetic dyes showed various peaks on HPLC chromatogram indicating different by-products formation. Finally, phytotoxicity analysis results revealed that the by-products of synthetic dyes (formed during decolorization) showed less toxicity against Phaseolus mungo compared to untreated synthetic dyes.
“…The analysis showed the occurrence of new peaks with disappearance of the control peaks in malachite green and methylene blue confirming effective decolorization by laccase whereas in case of congo red decolorization, appearance of new peaks along with detainment of control peak indicate slow and incomplete decolorization. The dye decolorization was supported with the help of HPLC analysis by various researchers earlier [23], [51], [52]. However, further study on identification of metabolites formed during dye decolorization is necessary.Figure 6HPLC analysis of Malachite green control (A), metabolites from Malachite green decolorization (B), Methylene blue control (C), metabolites from Methylene blue decolorization (D), Congo red control (E), metabolites from Congo red decolorization (F).…”
In this study we report the purification of laccase produced by Trichoderma harzianum strain HZN10 (using wheat bran under solid state fermentation) and its application in decolorization of synthetic dyes. Extracellular laccase was purified to homogeneity by DEAE-Sepharose and Sephadex G-100 chromatography with specific activity of 162.5 U/mg and 25-fold purification. Purified laccase was immobilized in various entrapments like calcium alginate, copper alginate, calcium alginate–chitosan beads and sol–gel matrix. Optimization results revealed that the laccase immobilized in sol–gel was optimally active in wide pH range (4.0–7.0) and thermo-stable (50–70 °C) than free enzyme which was optimum at 50 °C and pH 6.0. Kinetic analysis showed Km of 0.5 mM and 2.0 mM and Vmax of 285 U/mg and 500 U/mg by free laccase and sol–gel immobilized laccase respectively with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) [ABTS] substrate. Free and immobilized laccase was employed for decolorization of three different synthetic dyes (malachite green, methylene blue and congo red). High performance liquid chromatography (HPLC) analysis results revealed that approximately 100% of malachite green, 90% of methylene blue and 60% of congo red dyes at initial concentration of 200 mg/L were decolorized within 16, 18 and 20 h, respectively by laccase immobilized in sol–gel matrix in the presence of 1-hydroxybenzotriazole (HBT) mediator. During the decolorization all three synthetic dyes showed various peaks on HPLC chromatogram indicating different by-products formation. Finally, phytotoxicity analysis results revealed that the by-products of synthetic dyes (formed during decolorization) showed less toxicity against Phaseolus mungo compared to untreated synthetic dyes.
“…Heat-inactivated laccases were used as controls. The percentage of decolorization efficiency was calculated as follows:Decolorization (%) = (A i − A t )/A i × 100 where A i is initial absorbance of dyes, and A t is the absorbance of dye after each time point [66].…”
: Two laccase-encoding genes from the marine-derived fungus Pestalotiopsis sp. have been cloned in Aspergillus niger for heterologous production, and the recombinant enzymes have been characterized to study their physicochemical properties, their ability to decolorize textile dyes for potential biotechnological applications, and their activity in the presence of sea salt. The optimal pH and temperature of PsLac1 and PsLac2 differed in relation to the substrates tested, and both enzymes were shown to be extremely stable at temperatures up to 50 °C, retaining 100% activity after 3 h at 50 °C. Both enzymes were stable between pH 4–6. Different substrate specificities were exhibited, and the lowest Km and highest catalytic efficiency values were obtained against syringaldazine and 2,6-dimethoxyphenol (DMP) for PsLac1 and PsLac2, respectively. The industrially important dyes—Acid Yellow, Bromo Cresol Purple, Nitrosulfonazo III, and Reactive Black 5—were more efficiently decolorized by PsLac1 in the presence of the redox mediator 1-hydroxybenzotriazole (HBT). Activities were compared in saline conditions, and PsLac2 seemed more adapted to the presence of sea salt than PsLac1. The overall surface charges of the predicted PsLac three-dimensional models showed large negatively charged surfaces for PsLac2, as found in proteins for marine organisms, and more balanced solvent exposed charges for PsLac1, as seen in proteins from terrestrial organisms.
“…The decolorization in percentage for both dyes was determined, according to the equation [ 28 ] where A 0 is the initial absorbance of untreated dye solutions (control) and A t is the absorbance of dye solutions after enzymatic treatment.…”
Two anthraquinonic dyes, C.I. Acid Blue 225 and C.I. Acid Violet 109, were used as models to explore the feasibility of using the horseradish peroxidase enzyme (HRP) in the practical decolorization of anthraquinonic dyes in wastewater. The influence of process parameters such as enzyme concentration, hydrogen peroxide concentration, temperature, dye concentration, and pH was examined. The pH and temperature activity profiles were similar for decolorization of both dyes. Under the optimal conditions, 94.7% of C.I. Acid Violet 109 from aqueous solution was decolorized (treatment time 15 min, enzyme concentration 0.15 IU/mL, hydrogen peroxide concentration 0.4 mM, dye concentration 30 mg/L, pH 4, and temperature 24°C) and 89.36% of C.I. Acid Blue 225 (32 min, enzyme concentration 0.15 IU/mL, hydrogen peroxide concentration 0.04 mM, dye concentration 30 mg/L, pH 5, and temperature 24°C). The mechanism of both reactions has been proven to follow the two substrate ping-pong mechanism with substrate inhibition, revealing the formation of a nonproductive or dead-end complex between dye and HRP or between H2O2 and the oxidized form of the enzyme. Both chemical oxygen demand and total organic carbon values showed that there was a reduction in toxicity after the enzymatic treatment. This study verifies the viability of use of horseradish peroxidase for the wastewaters treatment of similar anthraquinonic dyes.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
