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 chromatog-raphy, on the CPOD enzyme was investigated. The Ki and IC50 values for this enzyme were calculated as 0.113 & PLUSMN; 0.012 mM and 0.196 & PLUSMN; 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 mgL1. 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.& COPY; 2023, The Society for Biotechnology, Japan. All rights reserved.

Automated summary

Enzymatic decolorization of industrial azo dyes in wastewater using peroxidases has gained increased interest. This study investigated the redox-mediated decolorization of Methylene Blue and Congo Red azo dyes using cauliflower peroxidase (CPOD) purified with a 4-amino 3-bromo 2-methyl benzohydrazide molecule. The molecule showed an inhibitory effect on the CPOD enzyme.