Laccase immobilization in polyelectrolyte multilayer membranes for 17α-ethynylestradiol removal: Biocatalytic approach for pharmaceuticals degradation

Abstract

Enzymatic membrane reactors equipped with multifunctional biocatalytic membranes are promising and sustainable alternatives for removal of micropollutants, including steroid estrogens, under mild conditions. Thus, in this study an effort was made to produce novel multifunctional biocatalytic polyelectrolyte multilayer membranes via polyelectrolyte layer-by-layer assembly with laccase enzyme immobilized between or into polyelectrolyte layers. In this study, multifunctional biocatalytic membranes are considered as systems composed of commercially available filtration membrane modified by polyelectrolytes and immobilized enzymes, which are produced for complex treatment of water pollutants. The multifunctionality of the proposed systems is related to the fact that these membranes are capable of micropollutants removal via simultaneous catalytic conversion, membrane adsorption and membrane rejection making remediation process more complex, however, also more efficient. Briefly, cationic poly-l-lysine and polyethylenimine as well as anionic poly(sodium 4-styrenesulfonate) polyelectrolytes were deposited onto NP010 nanofiltration and UFX5 ultrafiltration membranes to produce systems for removal of 17α-ethynylestradiol. Images from scanning electron microscopy confirm effective enzyme deposition, whereas results of zeta potential measurements indicate introduction of positive charge onto the membranes. Based on preliminary results, four membranes with over 70%, activity retention produced using polyethylenimine in internal and entrapped mode, were selected for degradation tests. Systems based on UFX5 membrane allowed over 60% 17α-ethynylestradiol removal within 100 min, whereas NP010-based systems removed over 75% of estrogen within 150 min. Further, around 80% removal of 17α-ethynylestradiol was possible from the solutions at concentration up to 0.1 mg/L at pH ranging from 4 to 6 and at the pressure up to 3 bar, indicating high activity of the immobilized laccase over wide range of process conditions. Produced systems exhibited also great long-term stability followed by limited enzyme elution from the membrane. Finally, removal of over 70% and 60% of 17α-ethynylestradiol, respectively by NP010 and UFX5 systems after 8 cycles of repeated use indicate high reusability potential of the systems and suggest their practical application in removal of micropollutants, including estrogens.

Keywords: 17α-ethynylestradiol; Biodegradation; Enzyme immobilization; Laccase; Layer-by-layer; Polyelectrolyte multilayer membranes.