Immobilization of a crude extract of laccase from T. hirsute Bm2 on copper alginate for environmental vinasse remediation

Sugarcane vinasses are environmentally aggressive effluents because of their high phenolic content and the presence of other recalcitrant compounds. The aim of this work was to immobilize laccases from Trametes hirsute Bm2 in copper alginate spheres in order to oxidize phenols and remove color from vinasses. Respond Surface Methodology (RSM) was used to evaluate enzyme and CuSO$_4$ concentration during immobilization. Laccase activity retained in spheres was the response variable. Lacasse activity in spheres increased by 30 \% and the best efficiency in immobilization was 95\% with 300 mM of CuSO$_4$ and 150 U/mL of laccase. Vinasse treatment (10\%) with free laccase removed 40\% of phenols, and the addition of natural mediators increased up to 60\%. Immobilized enzymes were able to remove up to 68\% of total phenols. Synthetic and natural mediators were used in the immobilization process to improve the removal of phenols by laccases. However, mediators did not significantly improve the process. The biocatalizer was able to remove phenols during four cycles of treatment and the maximum decoloration was 75\%. These phenomena were attributed to both laccase activity and adsorption to the support. After treatment, a dark precipitate was observed, suggesting polymerizing activity for laccases. These results reveal that laccases immobilized on copper alginate are a feasible alternative for the treatment of sugarcane effluents.

Keywords: phenols, vinasse, laccase, immobilization, cupper alginate.

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