3D printed polylactide scaffolding for laccase immobilization to improve enzyme stability and estrogen removal from wastewater.

This study reports a biocatalytic system of immobilized laccase and 3D printed open-structure biopolymer scaffoldings. The scaffoldings were computer-designed and 3D printed using polylactide (PLA) filament. The immobilization of laccase onto the 3D printed PLA scaffolds were optimized with regard to pH, enzyme concentration, and immobilization time. Laccase immobilization resulted in a small reduction in reactivity (in terms of Michaelis constant and maximum reaction rate) but led to significant improvement in chemical and thermal stability. After 20 days of storage, the immobilized and free laccase showed 80% and 35% retention of the initial enzymatic activity, respectively. The immobilized laccase on 3D printed PLA scaffolds achieved 10% improvement in the removal of estrogens from real wastewater as compared to free laccase and showed the significant reusability potential. Results here are promising but also highlight the need for further study to improve enzymatic activity and reusability.