Search Results (727)

Volatile fatty acids (VFAs) are essential precursors in chemical synthesis for various chemicals, polymers, pharmaceuticals, and fragrance compounds. Acidogenic anaerobic digestion (or arrested methanogenesis) is a promising method to stabilize organic wastes and convert them to value-added products such as VFAs. However, the VFAs’ accumulation could in turn suppress the fermentation process through product inhibition and limit the titer of VFA in the digestate. Therefore, in situ separation and recovery of VFAs from the fermentate is crucial to constructing an effective continuous VFA-producing system. Recent research has been dedicated to addressing these issues and advancing the utilization of biobased VFAs, particularly through process-intensified strategies employing novel green solvents such as natural deep eutectic solvents. Furthermore, in situ conversion of VFAs into esters is another potential strategy for VFA removal. However, VFA esterification in an aqueous medium is challenging due to the abundant water driving the reaction toward hydrolysis. Recent advances in free or immobilized enzyme catalysis in solvents have demonstrated improved ester yield by providing a hydrophobic space for the esterification reaction in aqueous solution. In this review, we present an overview of critical aspects on the state-of-the-art of green solvent-based process intensification strategies, including feedstock selection and pretreatment, operating condition optimization, advances in membrane- and solvent-based recovery methods, and biocatalytic in situ esterification. Lastly, we provide perspectives toward cost-effective, continuous, high-solid, environmental-benign, and industrial-relevant VFA production applications. Full article

Hydrogel-based optical sensors have emerged as a versatile class of analytical materials that combine soft-matter processability, tunable network chemistry, and compatibility with luminescent, colorimetric, photonic, and hybrid transduction strategies. Progress in the field is driven not by a single sensing mechanism, but by the convergence of key advances in material functionalization, embedded selectivity, operation across diverse sample matrices, mechanical and analytical robustness, and usability beyond the laboratory. Current systems include framework-integrated, nanoparticle-doped, probe-functionalized, photonic-crystal, enzyme-immobilized, and device-coupled hydrogels, reflecting growing architectural diversity and application-oriented engineering. Selectivity has likewise advanced from basic interferent screening to recognition-specific, imprinted, and pattern-discriminative formats suited to complex environmental, food, biological, and wearable settings. Evidence of stability, reusability, and deformation tolerance further suggests that many platforms are moving beyond proof-of-concept demonstrations toward credible real-world operation. At the same time, translational priorities such as portability, smartphone readout, implantable and epidermal formats, and multifunctionality spanning antimicrobial action, adsorption, anti-counterfeiting, and device integration are becoming increasingly prominent. Together, these trends show that hydrogel-based optical sensing is maturing into a materially rich, application-responsive domain. The key challenge ahead is to unify materials design, selectivity control, durability, and deployability in standardized, reproducible, and clinically or environmentally credible sensing platforms. Full article

Hormones regulate numerous physiological processes and are essential for maintaining metabolic homeostasis. Accurate hormone quantification is crucial for the diagnosis and monitoring of endocrine and metabolic disorders. Electrochemical biosensors have recently emerged as promising platforms for hormone detection, offering simplicity, rapid response, cost-effectiveness, and high sensitivity compared to conventional techniques such as chromatography and mass spectrometry. This review summarizes the advances in electrochemical biosensors for detecting clinically relevant hormones, including cortisol, estrogen, progesterone, thyroid-stimulating hormone, parathyroid hormone, prolactin, and insulin, since 2010. Particular attention has been paid to developments in electrode modification strategies, including nanomaterials, redox enzymes, and novel recognition elements, which significantly improve the sensitivity and selectivity. These advances enable hormone detection at lower concentrations in various biological and environmental matrices. Despite these promising developments, challenges related to sensor stability, fabrication costs, and regeneration procedures limit their large-scale commercialization. Future research should focus on improving robustness, optimizing immobilization strategies, and integrating innovative materials to enhance the analytical performance. Continued collaboration among researchers, engineers, and healthcare professionals is essential. With ongoing technological progress, electrochemical biosensors are expected to play an important role in clinical diagnosis, point-of-care testing, and personalized medicine. Full article

Cysteine proteases (bromelain, ficin, and papain) are widely used in biotechnology and medicine, but their application is limited by rapid autolysis and oxidative inactivation. This study aimed to develop effective supports for these enzymes based on graft copolymers of sodium alginate and poly(N-vinylpyrrolidone) (SA-g-PVP) and to elucidate the structure–property relationships governing immobilization efficiency, catalytic activity, and storage stability. Copolymers were synthesized via radical solution polymerization under optimized conditions. Enzymes were immobilized by physical adsorption, and the resulting complexes were characterized by Fourier-transform infrared (FTIR) spectroscopy, protein content assays, proteolytic and amidase activity measurements, and molecular docking. The graft copolymer with a smaller particle size in solution provided a larger accessible surface area, leading to higher bromelain and papain loading. Ficin showed the opposite trend due to its unique surface amino acid composition. Immobilization dramatically increased storage stability: half-life values for bromelain, ficin, and papain reached up to 20, 14, and 14 days, respectively, compared to 1–3 days for the free enzymes. Molecular docking revealed that the dense polymer shell stabilizes the enzyme tertiary structure by forming multiple contacts with internal cavities and tunnels, thereby preventing autolysis and conformational unfolding. Collectively, these findings demonstrate that SA-g-PVP copolymers are promising, non-toxic supports for cysteine proteases, with ficin showing up to 100% activity recovery, making them suitable for food, cosmetic, and biomedical applications. Full article

The use of hydrolytic enzymes is one of the most promising methods for combating bacterial biofilms. However, the use of native enzymes is limited by the rapid loss of activity under unfavorable conditions. Immobilization of enzymes on carbon nanoparticles enhances their stability, allows for biocatalyst reuse, and creates a synergistic effect due to the intrinsic antimicrobial properties of the nanomaterials. The aim of this investigation was to create and comparatively analyze conjugates of acid and alkaline proteases with carboxylated multiwalled carbon nanotubes (MWCNTs-COOH) and to assess their effect on the formation and destruction of E. coli VKM B-3858D biofilms. The immobilization efficiency and kinetics of enzyme adsorption on the support were quantified by determining the protein concentration using the Bradford assay. The morphology and dispersion of the resulting conjugates were analyzed using atomic force microscopy (AFM). Protease activity was determined by a modified Anson method using the Folin–Ciocalteu reagent. Biofilm biomass was determined using crystal violet staining. The binding efficiency of the acid protease to MWCNTs-COOH was shown to reach 93%, which is significantly higher than that of the alkaline protease. The highest degree of immobilization was observed at a protein concentration of 117–338 μg/mL (10–20 mg/mL of the enzyme preparations). The interaction of the acid protease with the carbon nanoparticles increased dispersion, reducing the size of aggregates from ~1 μm to ~68 nm. As a result, acid protease conjugates with MWCNTs-COOH significantly reduced the biofilm biomass compared to both the enzyme-free control and the native enzyme. Alkaline protease, unlike the acid protease, destroys mature biofilms, and immobilization on MWCNTs-COOH enhances this ability. Native alkaline protease and acid protease conjugates with MWCNTs-COOH are effective in combating the biofilm formation of Gram-negative bacteria, while alkaline protease conjugates are suitable for disrupting mature biofilms. Full article

This work introduces a textile-based platform for biocatalysis by integrating a copper-based hybrid domain onto aminated/amidine-functionalized acrylic textile (TAC–Cu), producing a functional bio-textile capable of high-performance enzyme immobilization. The textile substrate was chemically modified with ethylenediamine to generate amine/amidine-type functional groups, enabling in situ formation of copper-based hybrid structures through either a conventional solvothermal approach or a plant-mediated route employing Costus speciosus extract. The green-synthesized TAC–Cu composite exhibited superior structural uniformity, improved porosity, and enhanced surface chemistry, resulting in a higher horseradish peroxidase (HRP) immobilization yield (92%) compared with the chemically synthesized analogue. The resulting HRP-functionalized bio-textile demonstrated markedly improved catalytic behavior, including a reaction rate constant nearly twice that of the free enzyme, and strong operational robustness. As a technical textile engineered for environmental applications, the composite achieved 90% bisphenol A (BPA) removal within 90 min and retained substantial enzymatic activity even at 80 °C, whereas free HRP was almost fully deactivated. Overall, this study highlights the potential of eco-engineered TAC–Cu materials as a new class of functional and sustainable bio-textiles, combining enzyme stabilization, high catalytic efficiency, and suitability for wastewater treatment and other technical textile applications. Full article

Enzyme-Induced Carbonate Precipitation (EICP) represents a sustainable advancement in geotechnical engineering for stabilizing fine-grained soils (e.g., silt). Utilizing plant-derived urease (~12 nm) to catalyze urea hydrolysis, this technique generates calcium carbonate (CaCO₃) for soil reinforcement. Unlike Microbially Induced Carbonate Precipitation (MICP), EICP overcomes microbial size constraints (0.5–3 µm) by penetrating soil micropores, enabling uniform cementation. Its innovative single-phase low-pH method achieves >98% calcium conversion efficiency, yielding 6.41 MPa unconfined compressive strength (UCS) in sand—a 92.97% improvement over MICP. EICP demonstrates versatility: enhancing soil strength (up to 650% for silt), erosion resistance (wind erosion modulus increased ~20-fold), anti-seepage performance (permeability reduced from 10⁻⁶ to <10⁻⁹ cm/s), and heavy metal immobilization (>99%). However, challenges include unstable crystal morphologies (e.g., excessive vaterite), urease stability/cost constraints, and environmental concerns related to NH₃ emissions from urea hydrolysis. The manuscript acknowledges these emissions’ impacts and introduces mitigation strategies: ammonia capture technologies, optimized dosing protocols, and exploration of alternative N-sources. Long-term durability data under complex field conditions remain insufficient. Ongoing research addresses these gaps through nucleating agents (dried skim milk, biochar), enzyme immobilization, process optimization, and byproduct treatment. As a low-carbon technology with targeted mitigation measures, EICP advances environmentally conscious soil stabilization practices. This study presents a comparative narrative analysis of EICP’s performance and challenges, integrating laboratory findings and field applications. Full article

Plastic pollution caused by poly(ethylene terephthalate) (PET) highlights the urgent need for efficient biodegradation strategies. However, PET hydrolases such as DuraPETase typically exhibit limited substrate affinity for PET and insufficient operational stability. Although conventional immobilization improves enzyme stability, it often compromises catalytic activity. Here, we design a PET-targeting, orientation-controlled immobilization strategy that overcomes this traditional trade-off and enables efficient PET biodegradation. Guided by rational structural analysis, three Cys variants (R53C, R59C, R224C) were engineered for site-specific covalent attachment to a PDA@SiO₂ support with inherent PET-binding capability. The resulting conjugates (Dura_R53C-PDA@SiO₂, Dura_R59C-PDA@SiO₂, and Dura_R224C-PDA@SiO₂) displayed distinct catalytic and stability profiles. Among them, Dura_R59C-PDA@SiO₂ achieved the optimal balance between activity and stability, retaining kinetic properties comparable to the free enzyme and maintaining 87.6% residual activity after 2 h at 80 °C. Water contact angle measurements confirmed its PET-targeting behavior, as evidenced by the reduction in the PET contact angle from 85° to 45°. In 10-day degradation assays at 50 °C, Dura_R59C-PDA@SiO₂ released a total of 4865.32 μM degradation products, representing a 2.37-fold increase relative to free DuraPETase. These findings demonstrate an effective strategy for industrial enzymatic PET degradation and recycling. Full article

The microenvironment of the porous channels in enzyme immobilization carriers critically determines the catalytic performance of immobilized enzymes. In this study, we systematically tuned the hydrophobicity/hydrophilicity of the channel microenvironment of hydrogen-bonded organic frameworks (HOFs) by introducing four different substituents (-CH₃, -Cl, -F, -NH₂) at the 2-position of the phenyl ring of the HOF-101 monomer. These HOF-101 derivatives, which are isostructural to the parent HOF-101, were used to immobilize phosphotriesterase (PTE). The enzyme loading efficiencies ranged from 64.7% to 70.7%, indicating that the substituents had little effect on PTE binding, which primarily relies on carboxyl-residue interactions. Kinetic studies revealed that the hydrophilic -NH₂-functionalized HOF-101 (PTE@HOF-101-NH₂) exhibited the highest catalytic efficiency (1.43 × 10⁸ M⁻¹ s⁻¹), 2.27 times that of free PTE, while the hydrophobic -CH₃ analogue showed reduced activity. Notably, PTE@HOF-101-F demonstrated superior acid resistance (70% relative activity at pH 2) and long-term thermal stability (70% activity retention after 6 h at 70 °C), outperforming other derivatives. In contrast, PTE@HOF-101-NH₂ showed the highest activity under mild conditions but suffered from framework dissolution under prolonged harsh treatments. This work demonstrates that fine-tuning the HOF channel microenvironment is an effective strategy to enhance enzyme activity and stability, providing a platform for designing advanced immobilized enzyme systems. Full article

Two phenylacetaldehyde dehydrogenases, originating from Escherichia coli K-12 (FeaB-K-12) and Sphingopyxis fribergensis Kp5.2 (FeaB-Kp5.2), were immobilized on powdery silica carrier with various functionalization. First, the suitability of these carriers for application in combination with phenylacetaldehydes and phenylacetic acids was studied. Out of two carriers functionalized differently, mesoporous cellular foam, whose surface was modified with 3-glycidyloxypropyl groups (MCF-G), showed promising results. Hence, this carrier was further tested at 17 different immobilization conditions. Despite both enzymes showing high immobilization efficiency, the initial activities were relatively low compared to the free enzymes. Interestingly, the immobilized FeaB-Kp5.2 on MCF-G-Kw showed about 80% of retained activity after two months of incubation at 0 °C, indicating that the immobilization enhances the stability of this enzyme. In contrast, no changes in the temperature stability of FeaB-Kp5.2 due to immobilization could be noted. However, relative enzyme activities towards all three substituted phenylacetaldehydes could be increased by the immobilization to approximately 130%. The most active and stable powdery immobilizate was MCF-G-Kw-FeaB-Kp5.2 at pH 8. In addition, FeaB-Kp5.2 was also immobilized and tested on monolith silica carrier for continuous catalysis to produce phenylacetic acids. Full article

Nano–bio hybrid catalysts have emerged as a promising platform for sustainable energy conversion by integrating the high selectivity of enzymes with the structural robustness and conductivity of nanomaterials. In recent years, the growing demand for clean energy technologies has driven the development of biohybrid systems capable of efficient electron transfer, enhanced catalytic activity, and improved operational stability. This review comprehensively discusses the design principles, mechanistic foundations, and performance metrics of enzyme–nanomaterial interfaces for energy-related applications. We first outline the fundamentals of enzymatic redox catalysis and the limitations of free enzymes in practical systems. Subsequently, we examine the functional roles of nanomaterials including carbon-based materials, metal and metal oxide nanoparticles, and two-dimensional platforms such as MXenes in facilitating enzyme immobilization and promoting direct or mediated electron transfer. Special emphasis is placed on engineering strategies at the bio–nano interface, including immobilization techniques, surface functionalization, and structural tuning to optimize catalytic efficiency. The review further highlights representative hybrid systems based on laccase, glucose oxidase, peroxidase, and hydrogenase enzymes, and evaluates their applications in biofuel cells, solar–bio hybrid systems, green oxidation reactions, and self-powered biosystems. Stability challenges, deactivation mechanisms, and enhancement strategies such as polymer coatings, cross-linking, and nanoconfinement are critically analyzed. Finally, emerging directions including artificial enzymes, AI-guided catalyst design, and self-healing bioelectrodes are discussed to provide a forward-looking perspective on next-generation sustainable bioelectrocatalytic systems. Full article

This study presents a highly efficient and sustainable biocatalytic platform for bisphenol A (BPA) bioremediation through the covalent immobilization of laccase onto hierarchically functionalized cotton fibers. The immobilization strategy involved selective periodate oxidation of cellulose, grafting a hexamethylenediamine (HMDA) spacer arm, and glutaraldehyde activation, ensuring stable covalent attachment. Characterization via FTIR, SEM, and BET confirmed successful surface modification and high enzyme loading, achieving an immobilization yield of 90.5%. The immobilized laccase (CT-DA-HMD-Lac) exhibited significantly enhanced performance compared to the free enzyme, with a two-fold increase in maximum reaction velocity (V_max) and a 75% improvement in catalytic efficiency of action (V_max/K_m). Furthermore, the biocatalyst demonstrated superior robustness, maintaining high activity across broader pH and temperature ranges, and retaining 75% of its initial activity after 15 consecutive reusability cycles. Storage stability was also markedly improved, with 83% activity retention after 60 days. Practical application in BPA degradation showed 85% removal efficiency within 300 min, a 2.4-fold increase in the degradation rate constant over the free enzyme. These results highlight functionalized cotton as a promising, cost-effective, and scalable support for advanced enzymatic wastewater treatment and the remediation of persistent endocrine-disrupting chemicals. Full article

Biomineralization has recently emerged as a highly effective strategy for enzyme immobilization. Zeolitic imidazolate frameworks (ZIFs), a subclass of metal–organic frameworks (MOFs), are particularly attractive carriers due to their structural tunability and chemical stability. While ZIF-8 has been extensively studied, its denser and thermodynamically more stable analog ZIF-zni has received far less attention. In this work, we report the biomineralization of glucose oxidase (GOx) from Aspergillus niger within the ZIF-zni framework and systematically investigate the influence of zinc and imidazole (Im) concentration on immobilization performance. The optimized biocomposite, obtained at 10 mM Zn²⁺ and a Zn:Im ratio of 1:10, exhibited a specific activity of 2051 IU g⁻¹, which is more than twice the activity obtained for GOx@ZIF-8 in our previous study (874 IU g⁻¹). Furthermore, the GOx@ZIF-zni biocomposite demonstrated remarkable resistance to sodium dodecyl sulfate (SDS) and retained up to 50% of its activity after incubation at 65 °C for one hour. These results demonstrate that ZIF-zni is a highly promising carrier for enzyme immobilization and suggest that framework topology and synthesis conditions play a crucial role in determining the catalytic performance and stability of enzyme@MOF biocomposites. Full article

The enzymatic saccharification of cellulose remains a key step in biomass conversion processes, often influenced by enzyme stability, distribution, and accessibility at solid–liquid interfaces. Immobilization of cellulolytic enzymes on nanostructured supports has been proposed as a strategy to modulate catalytic behavior; however, the relationship between enzyme loading and catalytic response remains insufficiently understood. In this study, CuO-based nanobiocatalysts were prepared through controlled cellulase immobilization and systematically evaluated under defined experimental conditions. Structural and physicochemical characterization was performed using X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and integrated thermal analysis (TGA–DTG–DSC), enabling a comparative assessment of the analyzed systems. SEM analysis showed that the average particle diameter increased from 39.5 ± 14.8 nm (CuO nanoparticles) to 95.6 ± 21.8 nm (NPI10), 106.6 ± 27.7 nm (NPI15), and 113.5 ± 23.1 nm (NPI20), indicating progressive variations in particle organization with increasing enzyme loading. Catalytic performance was evaluated through enzymatic hydrolysis of cellulose filter paper as a model substrate, with products quantified by HPLC at a representative reaction time. The system prepared at lower enzyme loading (NPI10) exhibited product formation comparable to that of the free enzyme, with apparent average glucose formation values of 1.054 and 1.047 mg·mL⁻¹·h⁻¹, respectively. In contrast, higher immobilization levels were associated with reduced catalytic output. Across all systems, glucose was the predominant product, with negligible accumulation of intermediate oligomers under the evaluated conditions. These results indicate that increasing enzyme loading does not correspond to proportional increases in product formation and highlight the influence of enzyme distribution and accessibility within the system. The combined structural and catalytic observations provide a controlled framework for evaluating how immobilization conditions influence system behavior in nanobiocatalytic systems. Full article

Nickel (Ni) contamination threatens plant growth and ecosystem stability, and plant-growth-promoting rhizobacteria (PGPR) are sustainable bioremediation candidates. Here, we isolated and characterized a Ni-resistant PGPR strain, Microbacterium algeriense C14, from the rhizosphere of Zinnia elegans in Ni-contaminated soil. C14 exhibited exceptional Ni tolerance (up to 800 mg·L⁻¹), produced indole-3-acetic acid (IAA), and maintained pH homeostasis (8.3–8.7). XPS and XRD analyses confirmed a novel carboxylate-based precipitation mechanism: C14 secretes carboxyl-containing metabolites that coordinate with Ni²⁺ to form stable amorphous nickel–carboxylate complexes. Under Ni stress (50–600 mg·L⁻¹ for germination; 50–600 mg·kg⁻¹ soil for pot experiments), C14 inoculation increased the seed germination index by up to 47.3%, seedling root length by 36.9%, and mature plant aboveground fresh weight by 21.32%, while reducing plant Ni uptake by 38.7% (seedlings) and 49.9% (mature shoots). It also enhanced plant antioxidant-enzyme (SOD and POD) activities and soluble protein content, improved soil quality (pH +0.16–0.33 units, urease/acid phosphatase activities elevated), and reduced soil-available Ni by 23.7%. Additionally, C14 enriched Proteobacteria in the rhizosphere and modified microbial community structure. These results highlight M. algeriense C14 as a promising resource for Ni-contaminated soil remediation via integrated metal immobilization, growth promotion, and rhizosphere regulation. Full article