Purification and Characterization of Veratryl Alcohol Oxidase from Comamonas sp. UVS and Its Role in Decolorization of Textile Dyes

In the present work, we have purified veratryl alcohol oxidase (VAO) enzyme from Comamonas sp. UVS to evaluate its potential to decolorize textile dyes. VAO was purified (13.9 fold) by an ion exchange followed by the size exclusion chromatography. Molecular weight of the VAO was estimated to be about 66 kDa by SDS-PAGE. The optimum pH and temperature of oxidase were 30℃ and 65℃, respectively. VAO showed maximum activity with n-propanol among the various substrates (n-propanol, veratryl alcohol, L-dopa, tryptophan, etc.). Under standard assay conditions, K∧m value of the enzyme was 2.5 mM towards veratrole. The enzyme activity was completely inhibited by 0.5 mM sodium azide. L-cysteine, dithiothreitol, and the metal chelator, EDTA had a slight inhibitory effect. The purified enzyme was able to decolorize textile dyes, Red HE7B (57.5%) and Direct Blue GLL (51.09%) within 15 h at 40 ㎍/mL concentration. GC-MS analysis of the metabolites suggested oxidative cleavage and desulphonation of these dyes.