Investigation of reactive Blue 19 biodegradation and byproducts toxicity assessment using crude laccase extract from Trametes versicolor.

Crude laccase potency on biodegradation and detoxification of Reactive blue 19 (RB-19) were demonstrated, along with prediction of degradation mechanisms, pathways and byproducts analysis. Trametes versicolor, cultured on pampas grass inflorescence (Cortaderia selloana), yielded the best crude laccase activity (15.36 U/g). 10 U CLE activities demonstrated a biodegradation yield (85%) in 210 min, at pH 4, 50 °C and 200 mg/L RB-19 concentrations. Evolution of a brown color that absorbed maximally at 478 nm was observed during biodegradation. Two methods were adopted for byproducts extraction, three methods for toxicity analysis and four models for kinetic parameters (Km and Vmax) determination. 2-ethylanthracene, 2-hydroxycyclohexa-2,4-dien-1-one, 2(4-methylphenyl)-ethan-1-amine, 1-[6-hydroperoxy-4,5-bis(sulfooxy)oxan-3-yl]triaza-1,2-dien-2-ium, naphthalene-2,7-disulfonic acid and N-[(5-oxooxolan-2-yl)methyl]acetamide were detected as toxic byproducts. Brown color evolution was due to 1,1,1-triethyl-3-(methoxycarbonyl)-2,2-dioxo-2λ6-diazathian-1-ium (methoxycarbonyl sulfanyl-triethylammonium hydroxide) inner salt. Increase in color density (light to dark brown) was a function of byproduct(s) biodegradation and polymerization. RB-19 and byproduct acute toxicities were decreased significantly (98% - 6.91%). Kinetic parameters Km (18.05 mg/L) and Vmax (0.31 mg/L. min-1) from the four kinetic models demonstrated higher affinity of CLE to RB-19. CLE yielded a catalytic activity (Vmax/Km =0.017 min-1) demonstrating the flexibility of CLE active site to RB-19 binding over commercial laccase.