Search Results (312)

To meet the demand for rapid and accurate glucose determination in clinical diagnostics, food testing, and related fields, this study developed a high-performance electrochemical glucose biosensor based on multi-walled carbon nanotubes/Prussian blue/zeolitic imidazolate framework-8@glucose oxidase/chitosan (MWCNTs/PB/ZIF-8@GOx/CS). The MWCNTs/PB conductive network significantly accelerated electron transfer and catalytic activity, while the ZIF-8, with its regular pore structure and high specific surface area, provides an efficient microenvironment for the immobilization and conformational stabilization of glucose oxidase (GOx), thereby improving substrate diffusion and maintaining enzyme activity. The MWCNTs/PB/ZIF-8@GOx/CS sensor demonstrates excellent sensing performance, featuring a wide linear response to glucose concentrations ranging from 4.8 μM to 2.24 mM, a high sensitivity of 579.57 μA/mM/cm², and a low detection limit of 0.55 μM (S/N = 3). In addition, the sensor performs excellent repeatability (RSD = 1.49%) and retained 86.23% of its initial response after 3 weeks of storage at 4 °C, highlighting its strong potential for practical application in glucose detection. Full article

In vitro cultured biomass of Rindera graeca, a rare endemic plant, is an efficient renewable source of bioactive naphthoquinones, e.g., rinderol, a potential bioactive inducer of apoptosis in cancer cells. Bioengineering strategies, as biomass immobilization on functionalized biomaterial-based scaffolds, elicitation by chitosan, and in situ extraction of metabolites, are tested for intensifying naphthoquinones production in R. graeca hairy roots. The aim of the study was to investigate the effects of hybrid poly(lactic)–chitosan scaffolds on biomass proliferation and rinderol production in R. graeca hairy roots. Effects of chitosan origin (fungal or squid), molecular mass (350–1800 kDa), and concentration (up to 45%) in the developed hybrid scaffolds have been quantitatively identified, and the results were compared to the reference culture system containing an unmodified PLA-based construct. Applying PLA–chitosan scaffold containing 33% of fungal chitosan resulted in 635 times higher rinderol production (3660 µg g_DW⁻¹) than the application of reference scaffolds. Among the tested parameters, the chitosan concentration in the hybrid scaffolds revealed significant importance in rinderol production. To sum up, the developed hybrid PLA-chitosan scaffold may be recognized as a functional key element supporting the production of naphthoquinones in cultures of R. graeca biomass. Full article

Phosphatidylserine (PS) holds considerable importance in both the food and medical sectors; however, its biosynthesis is critically dependent on phospholipase D (PLD). The practical application of PLD is constrained by pronounced side reactions in its free form and by reduced selectivity when immobilized. To address these challenges, this study employed a sequential strategy involving bioimprinting to hyperactivate PLD, followed by microencapsulation via ionotropic gelation within an alginate–chitosan matrix. This approach induced conformational rigidification, enabling PLD to maintain its hyperactivated state in aqueous environments. Under optimal conditions, the encapsulation efficiency reached 78.56%, and the enzyme activity recovery achieved 105.27%. The immobilized bioimprinted PLD demonstrated exceptional catalytic performance, achieving a 94.68% PS yield within 20 min, which significantly surpassed that of free PLD (85.82% in 150 min) and non-imprinted immobilized PLD (90.34% in 60 min). This represents 7.27-fold and 2.14-fold efficiency improvements, respectively. Furthermore, the biocatalyst exhibited outstanding storage stability, thermal stability, and reusability (77.53% yield after 8 cycles). To our knowledge, this is the first report combining bioimprinting with alginate-chitosan microencapsulation via ionotropic gelation, which yielded remarkably enhanced PLD activity. These findings highlight the strong potential of this method for efficient PS production. Full article

Growing consumer awareness of the link between diet and health has increased interest in functional foods, including fermented juices. Grapefruit juice has potential health-promoting properties, but its bitter taste limits its acceptance by consumers. This study aimed to develop a fermentation process for debittering grapefruit juice at natural pH using Lacticaseibacillus rhamnosus and naringinase. Grapefruit juice was fermented with Lactic. rhamnosus using free naringinase and naringinase immobilized on carob gum and chitosan supports at 30 ± 0.2 °C for 72 h. Naringin concentration, bacterial cell count, total phenol content, organic acids, carbohydrates, antioxidant activity, and pH were analyzed. Naringinase immobilized on carob gum demonstrated the highest efficiency, hydrolyzing over 42% of naringin after 24 h (from 418.20 to 241.19 μg/mL). The free enzyme reduced the naringin concentration to 155.28 μg/mL after 48 h. The highest Lactic. rhamnosus cell count (2.05 × 10⁹ CFU/mL) was achieved with the free enzyme. Total phenol content decreased from 42.24 to 16.58 mg GAE/100 mL when using naringinase immobilized on chitosan. The combined use of naringinase and Lactic. rhamnosus enables the development of an integrated process that improves consumer acceptance with potential applications in the functional beverage industry. Full article

Foodborne illnesses remain a global challenge, requiring rapid and sensitive detection platforms. We developed a magnetosome-based electrochemical immunosensor for lipopolysaccharide (LPS) antigens from Escherichia coli and Salmonella typhimurium. Magnetosomes isolated from Magnetospirillum sp. RJS1 were characterized by HR-TEM and functionalized with antibodies (2 CFU mL⁻¹), with FTIR confirming successful conjugation. The antibody–magnetosome complexes were immobilized on a chitosan/glutaraldehyde-modified glassy carbon electrode. AFM revealed globular (200–700 nm) and island-like (1–3 µm) features after antigen binding. Electrochemical impedance spectroscopy showed stepwise increases in charge-transfer resistance upon electrode modification and antigen interaction. The sensor exhibited high sensitivity toward E. coli (3–7 CFU mL⁻¹) and Salmonella (3–8 CFU mL⁻¹), achieving an immune sensitivity of 36.24 Ω/CFU mL⁻¹ and a detection limit of 1 CFU mL⁻¹. These results demonstrate the potential of magnetosome-based immunosensors as portable, efficient platforms for the rapid detection of foodborne pathogens in real samples. Full article

The proteolytic fraction (P1G10) from Vasconcellea pubescens displays pharmacological activity in diverse therapeutic settings. It is responsible for antifungal activity against Botrytis cinerea, impairing its germination and the integrity of the plasma membrane. The application of P1G10 is limited by stability in aqueous environments, where proteases lose activity. In this study, we aim to stabilize the proteolytic fraction, by complexation, to preserve the enzymatic activity ensued by controlled release. The proportion of each polymer, and the established reaction sequence, is chitosan (CS) plus P1G10 and alginate (ALG) using ALG:CS mass ratio = 1.0. Scanning electron microscopy (SEM) of the product shows the ALG-CS-P1G10 complex displaying a rough surface contrasting with the smoother surface of the ALG-CS complex, likely induced by interactions between the protein and ALG-CS complex. The optimal amount of protein taken up by the complex under this condition was 13 mg, and the incorporation yield was 72%. The melting temperature (Tm) determined by differential scanning calorimetry (DSC) in ALG-CS increased from 80 °C to 86 °C for the biocatalyst ALG-CS-P1G10; this difference was probably induced by the interactions between P1G10 and ALG-CS. Fourier transform infrared spectrometry (FTIR) comparison between ALG-CS and ALG-CS-P1G10 shows two bands in the biocatalyst at 1601 and 1523 cm⁻¹, suggesting the presence of amine residues from P1G10 which is rich in lysine residues. The release of P1G10 from the complex was assessed by increasing the ionic strength in the media between 0.1 and 0.4 M NaCl. The results show that, at 0.3 M NaCl, the protein released after 8 h attained 70% and expressed enzymatic activity of 0.90 × 10⁻³ U/mg protein compared to the enzymatic activity from free P1G10 protein, which was 5.55 × 10⁻⁴ U/mg protein. Full article

Chitosanases are glycoside hydrolases (GHs) that catalyze the endo- or exo-type cleavage of β-1,4-glycosidic linkages in chitosan, enabling the selective production of chitooligosaccharides (COSs) with well-defined structures and diverse bioactivities. Owing to their substrate specificity and environmentally friendly catalytic action, chitosanases have garnered increasing attention as sustainable biocatalysts for COS production, with broad application potential in agriculture, food, medicine, and cosmetics. This review provides a comprehensive overview of recent advances in chitosanase research, focusing on the catalytic mechanisms and structure–function relationships that govern substrate selectivity and functional divergence across different GH families. Microbial diversity and heterologous expression systems for chitosanase production are discussed in parallel with biochemical characterization to support the rational selection of enzymes for specific biotechnological applications. Advances in protein engineering and computational approaches are highlighted as strategies to improve catalytic efficiency, substrate range, and stability. In addition, bioprocess optimization is addressed, with emphasis on fermentation using low-cost substrates and the application of immobilized enzymes and nano-biocatalyst systems for green and efficient COS production. Summarizing and discussing previous findings are essential to support future research and facilitate the development of next-generation chitosanases for sustainable industrial use. Full article

Organophosphorus pesticides (OPs) pose significant environmental and health risks due to their widespread use and toxicity, primarily by inhibiting acetylcholinesterase. Traditional detection methods are often slow and costly, highlighting the urgent need for advanced, sensitive, and accessible technologies. This study developed a highly sensitive electrochemical cholinesterase-inhibiting biosensor for OP pesticides, utilizing Ti₃C₂T_x MXene Quantum Dots (MQDs), which was synthesized via a hydrothermal method. The biosensor’s performance was characterized using electrochemical impedance spectroscopy, differential pulse voltammetry (DPV), and cyclic voltammetry. DPV proved to be the optimal technique, exhibiting an ultralow detection limit of 1 × 10⁻¹⁷ M and a wide linear range (10⁻¹⁴–10⁻⁸ M) for chlorpyrifos (a model OP) with an estimated inhibition constant of 62 nM. The biosensor demonstrated high selectivity for OPs (chlorpyrifos, acephate, glyphosate) over a non-target pyrethroid (permethrin), confirmed by distinct electrochemical signatures and compared to in vitro cholinergic activity assays in bean beetle homogenates. The enhanced performance is attributed to the high surface-to-volume ratio, quantum confinement effects, and superior conductivity of the MQDs, as well as the robust enzyme immobilization facilitated by glutaraldehyde cross-linking and a chitosan matrix. This work presents a promising platform for rapid, sensitive, and selective detection of OP pesticides, with potential applications in environmental monitoring and public health protection. Full article

7,12-Dimethylbenzo[a]anthracene (DMBA-7,12), a highly toxic and environmentally persistent polycyclic aromatic hydrocarbon (PAH), poses significant threats to marine biodiversity and human health due to its bioaccumulation through the food chain. Conventional chromatographic methods, while achieving comparable detection limits, are hindered by the need for expensive instrumentation and prolonged analysis times, rendering them unsuitable for rapid on-site monitoring of DMBA-7,12 in marine environments. Therefore, the development of novel, efficient detection techniques is imperative. In this study, we have successfully developed an electrochemical immunosensor based on a polydopamine (PDA)–chitosan (CTs) composite interface to overcome existing technical limitations. PDA provides a robust scaffold for antibody immobilization due to its strong adhesive properties, while CTs enhances signal amplification and biocompatibility. The synergistic integration of these materials combines the high efficiency of electrochemical detection with the specificity of antigen–antibody recognition, enabling precise qualitative and quantitative analysis of the target analyte through monitoring changes in the electrochemical properties at the electrode surface. By systematically optimizing key experimental parameters, including buffer pH, probe concentration, and antibody loading, we have constructed the first electrochemical immunosensor for detecting DMBA-7,12 in seawater. The sensor achieved a detection limit as low as 0.42 ng/mL. In spiked seawater samples, the recovery rates ranged from 95.53% to 99.44%, with relative standard deviations (RSDs) ≤ 4.6%, demonstrating excellent accuracy and reliability. This innovative approach offers a cost-effective and efficient solution for the in situ rapid monitoring of trace carcinogens in marine environments, potentially advancing the field of marine pollutant detection technologies. Full article

Water quality is essential for safeguarding human health and ensuring the stability of ecosystems. Nonetheless, the rising prevalence of emerging contaminants, particularly pharmaceutical compounds, has raised serious environmental concerns due to their bioactivity, widespread use, persistence, and potential toxicity. Among these, acetaminophen (paracetamol) is one of the most frequently detected pharmaceutical pollutants in aquatic environments. Among the various degradation strategies explored, biological methods, especially those involving white-rot fungi, have shown substantial promise owing to their production of ligninolytic enzymes capable of degrading complex pollutants. This study investigates the use of laccases from Ganoderma parvulum, covalently immobilized on chitosan microspheres, for acetaminophen degradation. The immobilization involved a 10% crosslinking agent, 60-min crosslinking time, and 10,000 U/L enzyme concentration, resulting in an immobilization efficiency of 123%, 203%, and 218%, respectively. The immobilized enzymes displayed enhanced stability across pH 3–8 and temperatures between 20 and 60 °C. Biodegradation assays achieved 97% acetaminophen removal within four hours. Nuclear Magnetic Resonance (¹H NMR and COSY) confirmed structural transformation. The enzymes also retained over 95% catalytic activity after multiple reuse cycles. These findings highlight the novel application of laccases as efficient and reusable biocatalysts for pharmaceutical pollutant removal, providing valuable insights into the mechanisms of enzymatic environmental remediation. Full article

This work aims to study the catalytic properties of Fe, Cr, and Cu catalysts deposited on chitosan–silica (SBA-15) composites in liquid phase oxidation of cyclohexane (CH) with H₂O₂ and photocatalytic hydrogen evolution reaction. The catalysts were obtained by consecutive adsorption of chitosan (CS) and metal ions (Fe³⁺, Cr³⁺, Cu²⁺) on SBA-15 at ambient conditions. Characterization of the catalysts by XRD, IR spectroscopy, XPS, TEM, SEM, etc., showed the CS and metal ion adsorption on the solid support. Modification with CS provided better immobilization of the metal ions on SBA-15. The synthesized catalysts demonstrated different performance in liquid phase oxidation of cyclohexane with H₂O₂ under mild conditions at 40 °C and atmospheric pressure. Cyclohexane conversion on Fe–CS/SBA-15 (18.5%) and Cr–CS/SBA-15 (21.6%) was higher than on Cu–CS/SBA-15 (9.3%). The influence of different conditions of the reaction such as time, temperature, catalyst dosage, substrate and oxidant ratio on cyclohexane conversion in the presence of the most efficient Cr–CS/SBA-15 catalyst was also studied. The optimal reaction conditions were found to be the following: duration of reaction—4 h, temperature of reaction—50 °C, m_cat—0.03 g, a substrate/H₂O₂ ratio of 1:3. In addition, Cr–CS/SBA-15 and Fe–CS/SBA-15 catalysts were studied in a photocatalytic H₂ evolution reaction. The Fe-containing catalyst demonstrated superior efficiency in photocatalytic H₂ evolution. The total volume of hydrogen produced within 3 h was 103 mL/g. Thus, this study demonstrates that chitosan possesses promising potential in the design of the supported catalysts for cyclohexane oxidation and photocatalytic hydrogen evolution reactions. Full article

D-allulose is a rare sugar with promising applications in food and health industries, owing to its low caloric value and multiple health benefits. In this study, we systematically investigated a thermostable D-allulose 3-epimerase (TcDAEase) from Thermogemmatispora carboxidivorans for food-compatible continuous production. The enzyme exhibited remarkable thermostability, with over 70% activity retained at 80 °C, and showed broad pH tolerance across the range of 8.0 to 13.0. Notably, TcDAEase exhibited high catalytic activity toward D-allulose and D-fructose even without the addition of metal ions. Moreover, food-grade Mg²⁺ was identified as enhancing enzyme activity by 14.3%, thus ensuring compliance with Generally Recognized as Safe (GRAS) standards for food applications. To improve industrial applicability, the enzyme was immobilized using a chitosan-diatomaceous earth (DE) matrix via three-step adsorption–crosslinking–embedding strategy. The immobilized TcDAEase achieved 48.7% ± 2.4% activity recovery and retained 90.3% ± 1.5% activity over seven reaction cycles. Furthermore, continuous production of D-allulose was realized in a packed-bed reactor, operating stably at 60 °C, pH 8.0 and 5 mM Mg²⁺ for 150 days, producing 756 kg of D-allulose with a conversion yield exceeding 89.7% of the theoretical maximum. Overall, this study provides a feasible strategy for the continuous and efficient production of high-value-added D-allulose in the food industry. Full article

This study reports, for the first time, the successful application of chitosan oligosaccharide (COS) and 2-hydroxyethyl cellulose (HEC) coatings on electrospun poly(3-hydroxybutyrate) (PHB) materials for the immobilization of non-conventional yeast strains with fungal biocontrol potential. The coatings enhanced the surface wettability of PHB fibers, facilitating efficient yeast adhesion and viability maintenance. Among the tested strains, Pichia acaciae YD6 was newly isolated and characterized, while Pichia fermentans YP6 and Zygosaccharomyces bailii YE1 had previously been identified as endophytic colonizers. All three strains demonstrated high adaptability, efficient immobilization, and antagonistic activity, confirming their potential for biocontrol applications. COS-coated PHB fibers promoted greater colony expansion than those coated with HEC. Antifungal assays of the yeast-containing biocarriers showed significant inhibition of F. graminearum growth. These findings underscore the potential of PHB-based fibrous materials as sustainable, bioactive carriers for yeast immobilization, with desirable biological properties. This approach offers a promising and eco-friendly strategy for pest control and bioactive agent delivery in agricultural applications. Full article

The low effectiveness of various brain cancer treatment methods is due to a number of significant challenges. Most of them are unable to penetrate the blood–brain barrier (BBB) when drugs are administered systemically through the bloodstream. Nanoscale particles play a special role among materials capable of binding drug molecules and successfully crossing the BBB. Biopolymeric nanoparticles (NPs) demonstrate excellent biocompatibility and have the remarkable ability to modify the environment surrounding tumor cells, thereby potentially improving cellular uptake of delivery agents. In our research, nanoscale polyelectrolyte complexes (PECs) ranging in size from 56 to 209 nm were synthesized by ionic interaction of the oppositely charged polysaccharides pectin and chitosan. The structural characteristics of these complexes were carefully characterized by infrared (FTIR) and Raman spectroscopy. The immobilization efficiency of antitumor drugs was comprehensively evaluated using UV spectrophotometry. The cytotoxicity of the NPs was evaluated in the U87-MG cell line. The preliminary data indicate a significant decrease in the metabolic activity of these tumor cells. Important details on the interaction of the NPs with an endothelial layer structurally similar to the BBB were obtained by simulating the BBB using a model based on human blood vessels. Our studies allowed us to establish a significant correlation between the kinetic parameters of drug immobilization and the ratio of biopolymer concentrations in the initial compositions, which provides valuable information for future optimization of drug delivery system design. Full article

In this study, we developed an innovative method for one-step enzyme purification and immobilization utilizing polysaccharide-based microspheres through a chitosan-binding module that mediated affinity adsorption. The chitosan-binding domain derived from Paenibacillus sp. IK-5 was genetically fused with multiple target enzymes (lysine decarboxylase, glutamate oxidase, and formate dehydrogenase), all of which were successfully expressed in soluble forms. Three distinct polysaccharide microspheres with optimized surface characteristics were engineered to facilitate the concurrent purification and immobilization of these fusion enzymes. Comprehensive characterization using organic elemental analysis, fluorescence microscopy, and thermogravimetric analysis confirmed the efficient immobilization of fusion enzymes. Remarkably, the immobilized enzymes demonstrated exceptional operational stability, maintaining over 80% of their initial catalytic activity after ten consecutive reuse cycles. This study establishes a robust and versatile platform for enzyme immobilization, providing significant advantages in biocatalyst engineering applications. Full article