This study assessed the immobilization of laccase on CPC silica beads and removal of two sulfonamides, sulfathiazole (STZ) and sulfamethoxazole (SMZ), using free and immobilized laccase. The results showed that free laccase (1Uml-1) removed 66% and 77% of SMZ and STZ, respectively, in the presence of 1-hydroxybenzotriazole (1mM) as a laccase mediator after incubation at 40°C for 1h. Maximum enzymatic removal of both applied sulfonamides was achieved at a temperature of 50°C and a pH of 5. At the elevated temperature of 70°C, immobilized laccase removed 71.7% of STZ and 53% of SMZ. After ten cycles of removal experiments, 63.3% and 82.6% of the initial activity of the immobilized laccase toward SMZ and STZ remained, respectively. A lower Kmvalue for STZ (0.056mM) compared to that of SMZ (0.096mM) confirmed a higher affinity of immobilized laccase toward STZ. A microtoxicity study of the inhibition of bacterial growth showed a decrease in toxicity of the laccase-treated sulfonamide solution.
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