Towards Multi-Analyte Detection with Field-Effect Capacitors Modified with Tobacco Mosaic Virus Bioparticles as Enzyme Nanocarriers

Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, tobacco mosaic virus (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO2-Ta2O5 layer structure for the sequential detection of penicillin and urea. The TMV-assisted bi-enzyme EISCAP biosensor exhibited a high urea and penicillin sensitivity of 54 and 85 mV/dec, respectively, in the concentration range of 0.1–3 mM. For comparison, the characteristics of single-enzyme EISCAP biosensors modified with TMV particles immobilized with either penicillinase or urease were also investigated. The surface morphology of the TMV-modified Ta2O5-gate was analyzed by scanning electron microscopy. Additionally, the bi-enzyme EISCAP was applied to mimic an XOR (Exclusive OR) enzyme logic gate.

Recent progress in magnetic nanoparticles and mesoporous materials for enzyme immobilization: an update

Enzymes immobilized onto substrates with excellent selectivity and activity show a high stability and can withstand extreme experimental conditions, and their performance has been shown to be retained after repeated uses. Applications of immobilized enzymes in various fields benefit from their unique characteristics. Common methods, including adsorption, encapsulation, covalent attachment and crosslinking, and other emerging approaches (e.g., MOFs) of enzyme immobilization have been developed mostly in recent years. In accordance with these immobilization methods, the present review elaborates the application of magnetic separable nanoparticles and functionalized SBA-15 and MCM-41 mesoporous materials used in the immobilization of enzymes.

ADAPTATION OF THE PROCEDURE OF CO-IMMOBILIZATION OF ENZYMES WITH DIFFERENT MODIFICATIONS OF ZEOLITES ON THE SURFACE OF CONDUCTOMETRIC TRANSDUCERS

A comparative study of different parameters of conductometric biosensors based on urease and glucose oxidase, co-immobilized with different types of zeolites. Urease immobilized on silicalite-2 was shown to have better performance than immobilized urease without zeolites. Conductometric biosensor with glucose oxidase co-immobilized with zeolite NH4+-Beta 25 had similar response values compared to immobilized enzyme without zeolite. Immobilization of zeolites NH4+-BEA 30 and H+-BEA 30 together with urease leads to an increase in the response of the biosensor, while the reproducibility of the signal remains unchanged. Biosensors with zeolites with a higher Si/Al ratio were characterized by increased signals. The use of zeolites modified with methylviologen and silver did not give a positive effect.

Production and Characterization of Cross-Linked Aggregates of Geobacillus thermoleovorans CCR11 Thermoalkaliphilic Recombinant Lipase

Immobilization of enzymes has many advantages for their application in biotechnological processes. In particular, the cross-linked enzyme aggregates (CLEAs) allow the production of solid biocatalysts with a high enzymatic loading and the advantage of obtaining derivatives with high stability at low cost. The purpose of this study was to produce cross-linked enzymatic aggregates (CLEAs) of LipMatCCR11, a 43 kDa recombinant solvent-tolerant thermoalkaliphilic lipase from Geobacillus thermoleovorans CCR11. LipMatCCR11-CLEAs were prepared using (NH4)2SO4 (40% w/v) as precipitant agent and glutaraldehyde (40 mM) as cross-linker, at pH 9, 20 °C. A U10(56) uniform design was used to optimize CLEA production, varying protein concentration, ammonium sulfate %, pH, glutaraldehyde concentration, temperature, and incubation time. The synthesized CLEAs were also analyzed using scanning electron microscopy (SEM) that showed individual particles of <1 µm grouped to form a superstructure. The cross-linked aggregates showed a maximum mass activity of 7750 U/g at 40 °C and pH 8 and retained more than 20% activity at 100 °C. Greater thermostability, resistance to alkaline conditions and the presence of organic solvents, and better durability during storage were observed for LipMatCCR11-CLEAs in comparison with the soluble enzyme. LipMatCCR11-CLEAs presented good reusability by conserving 40% of their initial activity after 9 cycles of reuse.

Immobilization of Enzymes by Polymeric Materials

Enzymes are the highly efficient biocatalyst in modern biotechnological industries. Due to the fragile property exposed to the external stimulus, the application of enzymes is highly limited. The immobilized enzyme by polymer has become a research hotspot to empower enzymes with more extraordinary properties and broader usage. Compared with free enzyme, polymer immobilized enzymes improve thermal and operational stability in harsh environments, such as extreme pH, temperature and concentration. Furthermore, good reusability is also highly expected. The first part of this study reviews the three primary immobilization methods: physical adsorption, covalent binding and entrapment, with their advantages and drawbacks. The second part of this paper includes some polymer applications and their derivatives in the immobilization of enzymes.

Development of Amino Acids Functionalized SBA-15 for the Improvement of Protein Adsorption

Ordered mesoporous materials and their modification with multiple functional groups are of wide scientific interest for many applications involving interaction with biological systems and biomolecules (e.g., catalysis, separation, sensor design, nano-science or drug delivery). In particular, the immobilization of enzymes onto solid supports is highly attractive for industry and synthetic chemistry, as it allows the development of stable and cheap biocatalysts. In this context, we developed novel silylated amino acid derivatives (Si-AA-NH2) that have been immobilized onto SBA-15 materials in biocompatible conditions avoiding the use of toxic catalyst, solvents or reagents. The resulting amino acid-functionalized materials (SBA-15@AA) were characterized by XRD, TGA, EA, Zeta potential, nitrogen sorption and FT-IR. Differences of the physical properties (e.g., charges) were observed while the structural ones remained unchanged. The adsorption of the enzyme lysozyme (Lyz) onto the resulting functionalized SBA-15@AA materials was evaluated at different pHs. The presence of different functional groups compared with bare SBA-15 showed better adsorption results, for example, 79.6 nmol of Lyz adsorbed per m2 of SBA-15@Tyr compared with the 44.9 nmol/m2 of the bare SBA-15.

Biocompatible cryogels: preparation and application

Polymer cryogels are very promising for producing functional materials. Their porous structure makes them indispensable for some areas of medicine, catalysis, and biotechnology. In this review we focused on methods for producing cryogels based on biopolymers, interpolyelectrolyte complexes of biopolymers, and composite cryogels based on them. First, the properties of cryogels and brief theoretical information about the production of cryogels based on biopolymers were considered. The second section summarizes the latest advances in the production of cryogels based on complexes of biopolymers and composite cryogels. The features of the synthesis and the factors affecting the final properties of materials were considered. In the final part the fields of application of cryogels of the considered types in biotechnology, catalysis and medicine were studied in detail. In biotechnology cryogels are used to immobilize molecules and cells, as a basis for cell growth, and as chromatographic materials for cell separation. In catalysis cryogels are used as a matrix for the immobilization of metal nanoparticles, as well as for the immobilization of enzymes. Biocompatible cryogels and their composites are widely used in medicine for bone and cartilage tissue regeneration, drug delivery, providing a long-term profile of drug release in the body.

Magnetic Nanomaterials as Biocatalyst Carriers for Biomass Processing: Immobilization Strategies, Reusability, and Applications

Environmental concerns, along with oil shortages, have increased industrial interest in biomass conversion to produce biofuels and other valuable chemicals. A green option in biomass processing is the use of enzymes, such as cellulases, hemicellulases, and ligninolytic (laccase and peroxidases), which have outstanding specificity toward their substrates and can be reused if immobilized onto magnetic nanocarriers. Numerous studies report the biocatalysts’ performance after covalent binding or adsorption on differently functionalized magnetic nanoparticles (MNPs). Functionalization strategies of MNPs include silica-based surfaces obtained through a sol–gel process, graphene oxide-based nanocomposites, polymer-coated surfaces, grafting polymer brushes, and others, which have been emphasized in this review of the immobilization and co-immobilization of enzymes used for biomass conversion. Careful analysis of the parameters affecting the performance of enzyme immobilization for new hybrid matrices has enabled us to achieve wider tolerance to thermal or chemical stress by these biosystems during saccharification. Additionally, it has enabled the application of immobilized laccase to remove toxic organic compounds from lignin, among other recent advances addressed here related to the use of reusable magnetic carriers for bioderived chemical manufacturing.