The widespread presence of estrogenic pollutants in aquatic environments poses a significant threat to ecosystems and human health, necessitating the development of efficient and sustainable removal technologies. This study aimed to develop a cost-effective biocatalyst for estrogen biodegradation using a fungal laccase. The enzyme was produced by the native strain Dichostereum sordulentum under semi-solid-state fermentation conditions optimized using a statistical Design of Experiments. The design evaluated carbon sources (glucose/glycerol), nitrogen sources (peptone/urea), inoculum size, and Eucalyptus dunnii bark as a solid support/substrate. The resulting laccase was entrapped within a hydrogel made of lignocellulosic biopolymers derived from a second-generation bioethanol by-product. Maximum laccase production was achieved with a high concentration of peptone (12 g/L), a low amount of bark (below 2.8 g), 8.5 g/L glucose and 300 mg/flask of inoculum. The subsequent immobilized laccase achieved 98.8 ± 0.5% removal of ethinylestradiol, outperforming the soluble enzyme. Furthermore, the treatment reduced the estrogenic biological activity by more than 170-fold. These findings demonstrate that the developed biocatalyst not only valorizes an industrial by-product but also represents an effective and sustainable platform for mitigating hazardous estrogenic pollution in water.

The widespread presence of estrogenic pollutants in aquatic environments poses a significant threat to ecosystems and human health, necessitating the development of efficient and sustainable removal technologies. This study aimed to develop a cost-effective biocatalyst for estrogen biodegradation using a fungal laccase. The enzyme was produced by the native strain Dichostereum sordulentum under semi-solid-state fermentation conditions optimized using a statistical Design of Experiments. The design evaluated carbon sources (glucose/glycerol), nitrogen sources (peptone/urea), inoculum size, and Eucalyptus dunnii bark as a solid support/substrate. The resulting laccase was entrapped within a hydrogel made of lignocellulosic biopolymers derived from a second-generation bioethanol by-product. Maximum laccase production was achieved with a high concentration of peptone (12 g/L), a low amount of bark (below 2.8 g), 8.5 g/L glucose and 300 mg/flask of inoculum. The subsequent immobilized laccase achieved 98.8 ± 0.5% removal of ethinylestradiol, outperforming the soluble enzyme. Furthermore, the treatment reduced the estrogenic biological activity by more than 160-fold. These findings demonstrate that the developed biocatalyst not only valorizes an industrial by-product but also represents an effective and sustainable platform for mitigating hazardous estrogenic pollution in water.