In recent years, groundwater contamination caused by dyes has become an important problem. They enter into wastewater as a result of the textile, automotive, or cosmetics industries. For this reason, new methods are being sought, which would aid at the removal of dye impurities with high efficiency and also would be relatively cheap. In the presented study synthesized TiO₂-ZrO₂ (with TiO₂:ZrO₂ molar ratio of 8:2) and TiO₂-ZrO₂-SiO₂ (with TiO₂:ZrO₂:SiO₂ molar ratio of 8:1:1) oxide materials were used as supports for enzyme immobilization. Effective synthesis of the carriers was confirmed by results of scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), low-temperature nitrogen sorption and Fourier transform infrared spectroscopy (FTIR). The materials achieve high immobilization efficiency of the laccases from Trametes versicolor (83% and 96% for TiO₂-ZrO₂-laccase and TiO₂-ZrO₂-SiO₂-laccase, respectively). The effect of selected dye concentrations, pH, temperature, and reusability were also tested. The obtained results showed that after removal of textile dyes, such as Alizarin Red S (ARS), Remazol Brilliant Blue R (RBBR), and Reactive Black 5 (RB5), under optimal process conditions, which were pH 5 and 25 °C, from dye solution of 5 mg/L degradation efficiency reached 100%, 91%, and 77%, respectively, suggesting synergistic mechanism of degradation by simultaneous sorption and catalytic action. Finally, reduction of chemical oxygen demand (COD) of the solution after treatment indicated lower mixture toxicity and effective dye degradation.

Nowadays, the increasing amounts of dyes present in wastewaters and even water bodies is an emerging global problem. In this work we decided to fabricate new biosystems made of nanofiltration or ultrafiltration membranes combined with laccase entrapped between polystyrene electrospun fibers and apply them for decolorization of aqueous solutions of three azo dyes, C.I. Acid Yellow 23 (AY23), C.I. Direct Blue 71 (DB71) and C.I. Reactive Black 5 (RB5). Besides effective decolorization of the permeate stream, the biosystems also allowed removal of dyes from the retentate stream as a result of enzymatic action. The effect of pH and applied pressure on decolorization efficiencies was investigated, and pH 5 and pressure of 2 bar gave the highest removal efficiencies of 97% for AY23 and 100% for both DB71 and RB5 from permeate solutions while decolorization of retentate for RB5 reached 65% under these conditions. Almost 100% decolorization of all dyes was achieved after three consecutive enzyme membrane cycles. Decolorization was shown to be due to the synergistic action of membrane separation and bioconversion. The biocatalytic action also enabled significant reduction of permeate and retentate toxicity, which is one of the biggest environmental health issues for these types of streams.

Nowadays, novel and advanced methods are being sought to efficiently remove dyes from wastewaters. These compounds, which mainly originate from the textile industry, may adversely affect the aquatic environment as well as living organisms. Thus, in presented study, the synthesized ZrO₂–SiO₂ and Cu²⁺-doped ZrO₂–SiO₂ oxide materials were used for the first time as supports for laccase immobilization, which was carried out for 1 h, at pH 5 and 25 °C. The materials were thoroughly characterized before and after laccase immobilization with respect to electrokinetic stability, parameters of the porous structure, morphology and type of surface functional groups. Additionally, the immobilization yields were defined, which reached 86% and 94% for ZrO₂–SiO₂–laccase and ZrO₂–SiO₂/Cu²⁺–laccase, respectively. Furthermore, the obtained biocatalytic systems were used for enzymatic decolorization of the Remazol Brilliant Blue R (RBBR) dye from model aqueous solutions, under various reaction conditions (time, temperature, pH). The best conditions of the decolorization process (24 h, 30 °C and pH = 4) allowed to achieve the highest decolorization efficiencies of 98% and 90% for ZrO₂–SiO₂–laccase and ZrO₂–SiO₂/Cu²⁺–laccase, respectively. Finally, it was established that the mortality of Artemia salina in solutions after enzymatic decolorization was lower by approx. 20% and 30% for ZrO₂–SiO₂–laccase and ZrO₂–SiO₂/Cu²⁺–laccase, respectively, as compared to the solution before enzymatic treatment, which indicated lower toxicity of the solution. Thus, it should be clearly stated that doping of the oxide support with copper ions positively affects enzyme stability, activity and, in consequence, the removal efficiency of the RBBR dye.

Enzymatic conversion of pharmaceutically active ingredients (API), using immobilized enzymes should be considered as a promising industrial tool due to improved reusability and stability of the biocatalysts at harsh process conditions. Therefore, in this study horseradish peroxidase was immobilized into sodium alginate capsules and then trapped into poly(vinyl chloride) electrospun fibers to provide additional enzyme stabilization and protection against the negative effect of harsh process conditions. Due to encapsulation immobilization, 100% of immobilization yield was achieved leading to loading of 25 μg of enzyme in 1 mg of the support. Immobilized in such a way, enzyme showed over 80% activity retention. Further, only slight changes in kinetic parameters of free (K_m = 1.54 mM) and immobilized horseradish peroxidase (K_m = 1.83 mM) were noticed, indicating retention of high catalytic properties and high substrate affinity by encapsulated biocatalyst. Encapsulated horseradish peroxidase was tested in biodegradation of two frequently occurring in wastewater API, sulfamethoxazole (antibiotic) and carbamazepine (anticonvulsant). Over 80% of both pharmaceutics was removed by immobilized enzyme after 24 h of the process from the solution at a concentration of 1 mg/L, under optimal conditions, which were found to be pH 7, temperature 25 °C and 2 mM of H₂O₂. However, even from 10 mg/L solutions, it was possible to remove over 40% of both pharmaceuticals. Finally, the reusability and storage stability study of immobilized horseradish peroxidase showed retention of over 60% of initial activity after 20 days of storage at 4 °C and after 10 repeated catalytic cycles, indicating great practical application potential. By contrast, the free enzyme showed less than 20% of its initial activity after 20 days of storage and exhibited no recycling potential.