Search Results (24)

In the presented study, a variety of hybrid and single nanomaterials of various origins were tested as novel platforms for horseradish peroxidase immobilization. A thorough characterization was performed to establish the suitability of the support materials for immobilization, as well as the activity and stability retention of the biocatalysts, which were analyzed and discussed. The physicochemical characterization of the obtained systems proved successful enzyme deposition on all the presented materials. The immobilization of horseradish peroxidase on all the tested supports occurred with an efficiency above 70%. However, for multi-walled carbon nanotubes and hybrids made of chitosan, magnetic nanoparticles, and selenium ions, it reached up to 90%. For these materials, the immobilization yield exceeded 80%, resulting in high amounts of immobilized enzymes. The produced system showed the same optimal pH and temperature conditions as free enzymes; however, over a wider range of conditions, the immobilized enzymes showed activity of over 50%. Finally, a reusability study and storage stability tests showed that horseradish peroxidase immobilized on a hybrid made of chitosan, magnetic nanoparticles, and selenium ions retained around 80% of its initial activity after 10 repeated catalytic cycles and after 20 days of storage. Of all the tested materials, the most favorable for immobilization was the above-mentioned chitosan-based hybrid material. The selenium additive present in the discussed material gives it supplementary properties that increase the immobilization yield of the enzyme and improve enzyme stability. The obtained results confirm the applicability of these nanomaterials as useful platforms for enzyme immobilization in the contemplation of the structural stability of an enzyme and the high catalytic activity of fabricated biocatalysts. Full article

The development of efficient strategies for wastewater treatment to remove micropollutants is of the highest importance. Hence, in this study, we presented a rapid approach to the production of biocatalytic membranes based on commercially available cellulose membrane and oxidoreductase enzymes including laccase, tyrosinase, and horseradish peroxidase. Effective enzyme deposition was confirmed based on Fourier transform infrared spectra, whereas results of spectrophotometric measurements showed that immobilization yield for all proposed systems exceeded 80% followed by over 80% activity recovery, with the highest values (over 90%) noticed for the membrane-laccase system. Further, storage stability and reusability of the immobilized enzyme were improved, reaching over 75% after, respectively, 20 days of storage, and 10 repeated biocatalytic cycles. The key stage of the study concerned the use of produced membranes for the removal of hematoporphyrin, (2,4-dichlorophenoxy)acetic acid (2,4-D), 17α-ethynylestradiol, tetracycline, tert-amyl alcohol (anesthetic drug), and ketoprofen methyl ester from real wastewater sampling at various places in the wastewater treatment plant. Although produced membranes showed mixed removal rates, all of the analyzed compounds were at least partially removed from the wastewater. Obtained data clearly showed, however, that composition of the wastewater matrix, type of pollutants as well as type of enzyme strongly affect the efficiency of enzymatic treatment of wastewater. Full article

Lignin-based spherical particles have recently gained popularity due to their characteristic and the usage of biopolymeric material. In this study, lignin-based spherical particles were prepared using choline chloride at different pH values, ranging from 2 to 10. Their dispersive, microstructural, and physicochemical properties were studied by a variety of techniques, including scanning electron microscopy, Fourier transform infrared spectroscopy, and zeta potential analysis. The best results were obtained for the particles prepared at pH 5 and 7, which had a spherical shape without a tendency to form aggregates and agglomerates. The lignin-based spherical particles were used for the immobilization of lipase, a model enzyme capable of catalyzing a wide range of transformations. It was shown that the highest relative activity of immobilized lipase was obtained after 24 h of immobilization at 30 °C and pH 7, using 100 mg of the support. Moreover, the immobilized lipase exhibited enhanced stability under harsh process conditions, and demonstrated high reusability, up to 87% after 10 cycles, depending on the support used. In the future, the described approach to enzyme immobilization based on lignin spheres may play a significant role in the catalytic synthesis of organic and fine chemicals, with high utility value. Full article

In this study, lipase from Aspergillus niger immobilized by physical immobilization by the adsorption interactions and partially interfacial activation and mixed physical immobilization via interfacial activation and ion exchange was used in the kinetic resolution of the ketoprofen racemic mixture. The FTIR spectra of samples after immobilization of enzyme-characteristic signals can be seen, and an increase in particle size diameters upon immobilization is observed, indicating efficient immobilization. The immobilization yield was on the level of 93% and 86% for immobilization unmodified and modified support, respectively, whereas activity recovery reached around 90% for both systems. The highest activity of immobilized biocatalysts was observed at pH 7 and temperature 40 °C and pH 8 and 20 °C for lipase immobilized by physical immobilization by the adsorption interactions and partially interfacial activation and mixed physical immobilization via interfacial activation and ion exchange, respectively. It was also shown that over a wide range of pH (from 7 to 10) and temperature (from 20 to 60 °C) both immobilized lipases retained over 80% of their relative activity, indicating improvement of enzyme stability. The best solvent during kinetic resolution of enantiomers was found to be phosphate buffer at pH 7, which obtained the highest efficiency of racemic ketoprofen methyl ester resolution at the level of over 51%, followed by enantiomeric excess 99.85% in the presence of biocatalyst obtained by physical immobilization by the adsorption interactions and partially interfacial activation. Full article

The occurrence of 17α-ethinylestradiol (EE2) in the environment and its removal have drawn special attention from the scientific community in recent years, due to its hazardous effects on human and wildlife around the world. Therefore, the aim of this study was to produce an efficient enzymatic system for the removal of EE2 from aqueous solutions. For the first time, commercial silica nanopowder and 3D fibrous chitinous scaffolds from Aplysina fistularis marine sponge were used as supports for horseradish peroxidase (HRP) immobilization. The effect of several process parameters onto the removal mechanism of EE2 by enzymatic conversion and adsorption of EE2 were investigated here, including system type, pH, temperature and concentrations of H₂O₂ and EE2. It was possible to fully remove EE2 from aqueous solutions using system SiO₂(HRP)–chitin(HRP) over a wide investigated pH range (5–9) and temperature ranges (4–45 °C). Moreover, the most suitable process conditions have been determined at pH 7, temperature 25 °C and H₂O₂ and EE2 concentrations equaling 2 mM and 1 mg/L, respectively. As determined, it was possible to reuse the nanoSiO₂(HRP)–chitin(HRP) system to obtain even 55% EE2 degradation efficiency after five consecutive catalytic cycles. Full article

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. Full article

Biotechnological use of probiotic microorganisms involves providing them with appropriate conditions for growth, but also protection against environmental changes caused by an exchange of the medium, isolation of metabolites, etc. Therefore, the research on effective immobilization of probiotic microorganisms should be focused in this direction. The present study aimed to evaluate the effectiveness of an innovative hybrid immobilization system based on electrospun nanofibers and alginate hydrogel. The analyses carried out included the study of properties of the initial components, the evaluation of the degree and durability of cell immobilization in the final material, and their survival under stress conditions. Effective binding of microorganisms to the hydrogel and nanofibers was confirmed, and the collected results proved that the proposed biocomposite is an efficient method of cell protection. In addition, it was shown that immobilization on electrospun nanofibers leads to the preservation of the highest cell activity and the least cell growth restriction as compared to free or lyophilized cells only. The completed research opens new perspectives for the effective immobilization of microorganisms of significant economic importance. Full article

The presented study deals with the fabrication of highly stable and active nanobiocatalysts based on Candida antarctica lipase B (CALB) immobilization onto pristine and poly(dimethylsiloxane) modified MWCNTs. The MWCNTs/PDMS nanocomposites, containing 40 wt.% of the polymer with two molecular weights, were successfully synthesized via adsorption modification. The effect of PDMS chains length on the textural/structural properties of produced materials was studied by means of the nitrogen adsorption–desorption technique, Raman spectroscopy, and attenuated total reflectance Fourier transform infrared spectroscopy. P-MWCNTs and MWCNTs/PDMS nanocomposites were tested as supports for lipase immobilization. Successful deposition of the enzyme onto the surface of P-MWCNTs and MWCNTs/PDMS nanocomposite materials was confirmed mainly using ATR-FTIR spectroscopy. The immobilization efficiency, stability, and catalytic activity of the immobilized enzyme were studied, and the reusability of the produced biocatalytic systems was examined. The presented results demonstrate that the produced novel biocatalysts might be considered as promising materials for biocatalytic applications. Full article

One of the directions of development in the textiles industry is the search for new technologies for producing modern multifunctional products. New solutions are sought to obtain materials that will protect humans against the harmful effects of the environment, including such factors as the activity of microorganisms and UV radiation. Products made of natural cellulose fibers are often used. In the case of this type of material, it is very important to perform appropriate pretreatment before subsequent technological processes. This treatment has the aim of removing impurities from the surface of the fibers, which results in the improvement of sorption properties and adhesion, leading directly to the better penetration of dyes and chemical modifiers into the structure of the materials. In this work, linen fabrics were subjected to a new, innovative treatment being a combination of bio-pretreatment using laccase from Cerrena unicolor and modification with CuO-SiO₂ hybrid oxide microparticles by a dip-coating method. To compare the effect of alkaline or enzymatic pretreatment on the microstructure of the linen woven fabrics, SEM analysis was performed. The new textile products obtained after this combined process exhibit very good antimicrobial activity against Candida albicans, significant antibacterial activity against the Gram-negative Escherichia coli and the Gram-positive Staphylococcus aureus, as well as very good UV protection properties (ultraviolet protection factor (UPF) > 40). These innovative materials can be used especially for clothing or outdoor textiles for which resistance to microorganisms is required, as well as to protect people who are exposed to long-term, harmful effects of UV radiation. Full article

Laccase enzyme from white-rot fungi is a potential biocatalyst for the oxidation of emerging contaminants (ECs), such as pesticides, pharmaceuticals and steroid hormones. This study aims to develop a three-step platform to treat ECs: (i) enzyme production, (ii) enzyme concentration and (iii) enzyme application. In the first step, solid culture and liquid culture were compared. The solid culture produced significantly more laccase than the liquid culture (447 vs. 74 µM/min after eight days), demonstrating that white rot fungi thrived on a solid medium. In the second step, the enzyme was concentrated 6.6 times using an ultrafiltration (UF) process, resulting in laccase activity of 2980 µM/min. No enzymatic loss due to filtration and membrane adsorption was observed, suggesting the feasibility of the UF membrane for enzyme concentration. In the third step, concentrated crude enzyme was applied in an enzymatic membrane reactor (EMR) to remove a diverse set of ECs (31 compounds in six groups). The EMR effectively removed of steroid hormones, phytoestrogen, ultraviolet (UV) filters and industrial chemical (above 90%). However, it had low removal of pesticides and pharmaceuticals. Full article

For the first time, 3D chitin scaffolds from the marine demosponge Aplysina archeri were used for adsorption and immobilization of laccase from Trametes versicolor. The resulting chitin–enzyme biocatalytic systems were applied in the removal of tetracycline. Effective enzyme immobilization was confirmed by scanning electron microscopy. Immobilization yield and kinetic parameters were investigated in detail, in addition to the activity of the enzyme after immobilization. The designed systems were further used for the removal of tetracycline under various process conditions. Optimum process conditions, enabling total removal of tetracycline from solutions at concentrations up to 1 mg/L, were found to be pH 5, temperature between 25 and 35 °C, and 1 h process duration. Due to the protective effect of the chitinous scaffolds and stabilization of the enzyme by multipoint attachment, the storage stability and thermal stability of the immobilized biomolecules were significantly improved as compared to the free enzyme. The produced biocatalytic systems also exhibited good reusability, as after 10 repeated uses they removed over 90% of tetracycline from solution. Finally, the immobilized laccase was used in a packed bed reactor for continuous removal of tetracycline, and enabled the removal of over 80% of the antibiotic after 24 h of continuous use. Full article

In this study, novel Ga₂O₃/lignin and ZrO₂/lignin hybrid materials were obtained and used as supports for the adsorption of the enzyme glucose oxidase (GOx). A biosensor system based on the hybrid supports was then designed to determine the concentration of glucose in various solutions. The obtained bioinspired platforms were analyzed to determine chemical and physical properties of the support structures. A determination was made of the effectiveness of the proposed method of immobilization and the quality of operation of the constructed glucose biosensor in electrochemical tests. To characterize the materials, Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), thermogravimetric analysis (TGA), electrokinetic (zeta) potential measurements, atomic force microscopy (AFM), particle size measurements (NIBS technique), and elemental analysis (EA) were used. In further research, glucose oxidase (GOx) was immobilized on the surface of the obtained functional Ga₂O₃/lignin and ZrO₂/lignin biomaterials. The best immobilization capacities—24.7 and 27.1 mg g⁻¹ for Ga₂O₃/lignin and ZrO₂/lignin, respectively—were achieved after a 24 h immobilization process. The Ga₂O₃/Lig/GOx and ZrO₂/Lig/GOx systems were used for the construction of electrochemical biosensor systems, in a dedicated carbon paste electrode (CPE) with the addition of graphite and ferrocene. Full article

The conversion of biomass components catalyzed via immobilized enzymes is a promising way of obtaining valuable compounds with high efficiency under mild conditions. However, simultaneous transformation of glucose and xylose into gluconic acid and xylonic acid, respectively, is an overlooked research area. Therefore, in this work we have undertaken a study focused on the co-immobilization of glucose dehydrogenase (GDH, EC 1.1.1.118) and xylose dehydrogenase (XDH, EC 1.1.1.175) using mesoporous Santa Barbara Amorphous silica (SBA 15) for the simultaneous production of gluconic acid and xylonic acid. The effective co-immobilization of enzymes onto the surface and into the pores of the silica support was confirmed. A GDH:XDH ratio equal to 1:5 was the most suitable for the conversion of xylose and glucose, as the reaction yield reached over 90% for both monosaccharides after 45 min of the process. Upon co-immobilization, reaction yields exceeding 80% were noticed over wide pH (7–9) and temperature (40–60 °C) ranges. Additionally, the co-immobilized GDH and XDH exhibited a significant enhancement of their thermal, chemical and storage stability. Furthermore, the co-immobilized enzymes are characterized by good reusability, as they facilitated the reaction yields by over 80%, even after 5 consecutive reaction steps. Full article

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. Full article