A novel peroxidase from runner bean (Phaseolus coccineus L.): Enhanced affinity purification, characterization, and dye decolorization activity.

Abstract

In this study, a novel runner bean peroxidase (RBP) was purified and characterized. Affinity-based purification was performed with newly synthesized disubstituted 4-aminobenzohydrazides. In the purification results, 253-fold was achievedwith a yield of 56.2%. Furthermore, molecular weight and enzyme purity were checked with the SDS-PAGE and observed a single band at 31.2kDa. Optimum conditions were determined as temperature=50°C, ionic strength=0.2M, and pH 7.0. Enzyme exhibited 31.2% of residual activity in the presence of 20% DMSO. Additionally, the redox-mediated decolorization effect of the enzyme was examined for Reactive Blue 19 and Acid Blue 25 dyes. As a result of 1-hr incubation, the enzyme removal activity of Reactive Blue 19 and Acid Blue 25 dyes was calculated as 47% and 57%, respectively. PRACTICAL APPLICATIONS: Peroxidases (PODs) ability to catalyze various redox reactions for many substrates makes them significant enzymes in industrial sectors. In our current report, a single-step strategy was developed and followed as an alternative to multi-step methods commonly used for the purification of PODs. During this process, high yield was achieved and the separation time was shortened. Also, the purification of RBP that can potentially supplant PODs used in the industrial applications was carried out for the first time. In addition, substrate specificity, catalytic behavior in water-miscible organic solvents, and dye bleaching activity of this enzyme have been determined to evaluate the utilization capacity in various processes.