Search Results (180)

Vibrio parahaemolyticus is the leading cause of bacterial diarrhea in humans from shellfish consumption. In Thailand, blood clam is a popular shellfish, but homemade cooking often results in insufficient heating. Therefore, consumers may suffer from food poisoning due to Vibrio infection. This study aimed to determine the effect of chitooligosaccharide conjugated with epigallocatechin gallate (COS-EGCG) at different concentrations (200 and 400 ppm) combined with high-voltage atmospheric cold plasma (HVACP) on inhibiting V. parahaemolyticus in vitro and in challenged blood clam meat. Firstly, HVACP conditions were optimized for gas composition and treatment time (20 and 60 s); a 70% Ar and 30% O₂ gas mixture resulted in the highest ozone formation and a treatment time of 60 s was used for further study. COS-EGCG conjugate at 400 ppm with HVACP (ACP-CE400) completely killed V. parahaemolyticus after incubation at 37 °C for 6 h. Furthermore, an antibacterial ability of ACP-CE400 treatment against bacterial cells was advocated due to the increased cell membrane damage, permeability, and leakage of proteins and nucleic acids. Scanning electron microscopy (SEM) showed cell elongation and pore formation, while confocal microscopy revealed disrupted biofilm formation. Additionally, the shelf life of challenged blood clam meat treated with ACP-CE400 was extended to nine days. SEM analysis revealed damaged bacterial cells on the meat surface after ACP-CE400 treatment, indicating the antibacterial activity of the combined treatment. Thus, HVACP combined with COS-EGCG conjugate, especially at a highest concentration (400 ppm), effectively inhibited microbial growth and extended the shelf life of contaminated blood clam meat. Full article

Chitooligosaccharides (COSs), degraded products of chitosan or chitin, are attracting growing interest owing to their low degree of polymerization (DP), high solubility, and prominent anti-inflammatory activity. However, the correlation between their structure and anti-inflammatory activities still needs to be explored. In this study, we use LPS-stimulated RAW 264.7 macrophages as an inflammatory model to systematically evaluate COS1–7 for their effects on inflammatory mediators and NF-κB signaling pathways. The results of Griess assay, ELISA, and real-time quantitative PCR show that COSs can inhibit the expression of NO, iNOS, and pro-inflammatory cytokines (IL-6, TNF-α, MCP-1 and IL-1β), thereby attenuating inflammatory signaling. Notably, chitohexaose (COS6) exhibits the most significant anti-inflammatory effect, reducing the mRNA levels of LPS-induced iNOS, IL-6, and IL-1β and the production of IL-6 and TNF-α by more than 50%. Transcriptome, western blotting, and real-time quantitative PCR analysis reveal that COSs can inhibit the activation of the NF-κB signal pathway by down-regulating TLR2 levels. Additionally, molecular docking confirms that COSs retard TLR2/4 dimerization and LPS recognition by TLR4, affecting downstream signaling cascades. In summary, this study provides a valuable insight into the potential anti-inflammatory mechanism of COSs and highlights the possible applications in human health promotion by modulating receptor-mediated signaling pathways. Full article

The depolymerization of chitosan using chitinolytic enzymes is one of the most promising approaches for the production of bioactive soluble chitooligosaccharides (COS) due to its high specificity, environmental safety, mild reaction conditions, and potential for development. However, the comparative efficacy of bacterial chitinases and chitosanases in terms of yield, solubility, and antimicrobial activity of produced COS remains understudied. In this work, chitinase (73 kDa) and chitosanase (40 kDa) from the strain Bacillus thuringiensis B-387 (Bt-387) were purified using various chromatographic techniques and compared by their action on chitosan (DD 85%). The molecular mass and structure of generated COS was determined using TLC, LC-ESI-MS, HP-SEC, and C¹³-NMR techniques. Chitosanase converted the polymer more rapidly to short COS (GlcN₂-GlcN₄), than chitinase, and was more specific in its action on mixed bonds between GlcN and GlcNAc. Chitosanase needed a noticeably shorter incubation time and enzyme–substrate ratio than chitinase for production of larger oligomeric molecules (Mw 2.4–66.5 and 15.4–77.7 kDa, respectively) during controlled depolymerization of chitosan. Moreover, chitosanase-generated oligomers demonstrate better solubility and a higher antifungal activity in vitro against the tested plant pathogenic fungi. These features, as well as the high enzyme production and its simplified purification protocol, make chitosanase B-387 more suitable for the production of antifungal chitooligomers than chitinase. Full article

Fresh wet vermicelli is highly susceptible to microbial contamination during storage as a result of its high moisture content and rich nutrients, which leads to spoilage and deterioration. In addition to exerting a great impact on the quality of the product, this results in significant economic losses and potential food safety risks. This work aimed to identify spoilage microorganisms via traditional culturing methods and molecular biology techniques. The effects of environmental factors such as temperature and pH on the growth and development of the dominant spoilage fungi were investigated, and the inhibitory effects of both chemical (potassium sorbate) and natural antimicrobial agents (chitooligosaccharides, chitosan, tea polyphenols, citric acid, and ε-polylysine hydrochloride) were evaluated. The results indicated that Penicillium crustosum was the major spoilage microorganism in fresh wet vermicelli, whose optimal growth temperature and pH was 28 °C and 7, respectively. While conidial germination began at 7 h, hyphal formation was only observed after 12 h. Moreover, the findings suggest that both natural and chemical antimicrobial agents can effectively inhibit the growth of P. crustosum, with ε-polylysine hydrochloride being the strongest antimicrobial agent. Overall, the findings of this study provide a scientific foundation for improving the preservation of fresh wet vermicelli, which is of great significance for extending its shelf life and enhancing food safety. Full article

Chitooligosaccharides (COS) exhibits good activity of inducing plant resistance, but the structure–activity relationship is still unclear. In this study, chitin oligosaccharides (CHOS) with a degree of polymerization (DP) of 2~6 were used as raw materials. Three deacetylases (NodB, VcCOD, and ArCE4A) were employed to prepare three different sequence-arranged COSs, namely N-COS, C-COS, and A-COS, and their structures were characterized by infrared spectroscopy, high-performance liquid chromatography, and mass spectrometry. Further studies were conducted on inducing the plant salt resistance of the three different sequence-arranged COSs on wheat seedlings. The results showed a sequence-dependent effect of COS inducing plant salt resistance. Among them, A-COS exhibited the best activity. When sprayed at a concentration of 10 mg/L on wheat seedlings under salt stress for 3 days, the leaf length of the wheat seedlings sprayed with A-COS was recovered, and the wet mass and dry mass were recovered by 20.40% and 6.64%, respectively. Following the enhancement of proline accumulation, the malondialdehyde content decreased by 34.75%, and the Na⁺/K⁺ ratio also exhibited a significant reduction, thereby alleviating salt stress-induced damage. This study was the first to demonstrate the effect of COS with specific sequences on inducing plant salt resistance, providing a theoretical basis for the development of a new generation of efficient COS plant biostimulator. Full article

The combined effects of chitooligosaccharide–epigallocatechin gallate conjugate (CEC) at different concentrations (1, 2, and 3%, w/w) and cold atmospheric plasma (CAP) on the depurated Asian green mussel edible portion (AGM-EP) were investigated during refrigerated storage for 15 days. Among all the treatments, the microbial counts, total volatile bases (TMA-N and TVB-N), and lipid oxidation of AGM-EP-treated 3% CEC in conjunction with CAP (CEC-3-CAP) were lower than the other samples during 15-day storage (p < 0.05). Total viable bacteria (6.16 log CFU/g sample), psychrotrophic bacteria (3.24 log CFU/g sample), Vibrio spp. (2.47 log CFU/g sample), presumptive Pseudomonas (5.93 log CFU/g sample), and H₂S-producing bacteria (5.05 log CFU/g sample) counts of the CEC-3-CAP were lower than samples treated with 1 and 2% (w/w) CEC on day 15, as well as samples solely treated using CAP during refrigerated storage, irrespective of storage time. Additionally, CEC-3-CAP had significantly lower lipid oxidation (PV: 8.36 mg cumene hydroperoxide/kg sample and TBARS: 2.65 mg MDA/kg sample) as compared to those without CEC added and other samples (p < 0.05). The incorporation of CEC effectively mitigated lipid oxidation as supported by lower reduction of PUFAs in AGM-EP. Moreover, on day 0, no significant differences were observed in cooking loss or textural parameters (firmness and toughness) among the treatments (p > 0.05). However, as storage progressed, cooking loss increased in the CEC-3-CAP sample, while a noticeable decline in firmness and toughness was recorded (p < 0.05). This further attributed to the lower likeness attained for CAP-3-CAP on day 12, but the score was higher than the acceptable limit (5.0). Therefore, CAP together with CEC is a promising technology to prolong the shelf-life of depurated AGM-EP by at least 9 days as compared to the control (3 days), but it certainly needs further studies for the retention of textural properties and sensorial attributes. Full article

The storage duration and hatchability of eggs largely depend on the lysozyme content in egg whites; therefore, determining the lysozyme status is important for characterizing their quality. For the first time, a fast and accurate method for determining the active lysozyme in egg whites has been proposed to establish the lysozyme status of eggs using the fluorescence polarization assay and synthetic chitooligosaccharide conjugates with a fluorescent label without sample preparation. The egg whites of hens, black hens, chukars, quails, ducks, geese, turkeys, peacocks, and ostriches were studied. Samples of egg whites from hens, black hens, chukars, and quails demonstrate the possibility of measuring the lysozyme activity. Samples of hen and black hen eggs from a farm showed approximately the same enzymatic activity of lysozyme. A relatively higher enzymatic activity was demonstrated by the samples from quail egg whites; however, a wide range of data was observed among the eggs. Chitooligosaccharide conjugates demonstrate that they bind only to C-type lysozyme, and no interaction with G-type lysozyme has been shown. Lysozyme activity in the egg whites of duck, goose, turkey, peacock, and ostrich eggs has not been detected by using the obtained chitooligosaccharide tracers, which may be related to the structural features of lysozyme in different bird species. Thus, the method of fluorescence polarization (FP), using fluorescently labeled chitopentaoside to determine the lysozyme status, can be used to characterize hen, black hen, chukar, and quail eggs, which will allow for the selection of a batch of eggs with a high content of active lysozyme, for example, for long-term storage. Full article

Natural pigments like betacyanins are highly unstable under heat, light, acid, and alkaline conditions. Yeast protein (YP) is a promising substitute protein, while chitooligosaccharides (COS) are water-soluble alkaline polysaccharides. Water-in-oil-in-water (W₁/O/W₂) emulsions, with two-membrane, three-phase structure, can serve as effective carriers for stabilizing pigments. In this study, YP-COS complexes formed through electrostatic interactions were used as hydrophilic emulsifiers to create betacyanin-coated W₁/O/W₂ emulsions. The W₁/O colostrum was designed to make up 30%, 70%, and 90% of the emulsion (v/v)and the W₂ was designed by the complexes with three concentrations of YP (2%, 1.25% and 0.5%, w/v)-COS (6%, 3.75% and 1.5%, w/v). The optimal formulation was determined through comprehensive evaluation of micromorphological characteristics, particle size, zeta potential and creaming index, ultimately yielding a system comprising YP (2%)-COS (6%) and 90% W₁/O colostrum. Moreover, the W₁/O/W₂ emulsion system significantly improved the betacyanins retention under thermal treatment, photolytic exposure, pH gradients, and extended storage compared to the betacyanin aqueous solution (p < 0.05). In vitro digestion tests showed the emulsion retained 58.39% of betacyanins, while the betacyanin aqueous solution retained only 41.42%, demonstrating the emulsion’s ability to delay the betacyanins release, offering new insights for using YP-COS complexes in food production and other fields. Full article

Termitomyces, a rare edible fungus with both nutritional and medicinal value, has garnered significant attention for its antioxidant properties. This study aims to elucidate the effects of various nutritional components on the antioxidant activity of Termitomyces. Through assays including FRAP, DPPH, ABTS, and •OH scavenging activity, strain XNQL025, which exhibits high antioxidant activity, was identified. Subsequent optimization of culture medium components using single-factor experiments and response surface methodology revealed that aspartic acid (Asp) significantly enhances the antioxidant capacity of this strain. Transcriptomic analysis showed that Asp activates key pathways, including glycolysis/gluconeogenesis, propanoate metabolism, amino sugar and nucleotide sugar metabolism, valine–leucine–isoleucine biosynthesis, and tryptophan metabolism, along with modulating the peroxisome and mitogen-activated protein kinase (MAPK) signaling pathways. These regulatory actions promote the synthesis of antioxidant compounds and establish a multi-layered antioxidant defense system comprising enzymatic (catalase) and non-enzymatic (leucine/chitooligosaccharides) components. The synergistic interaction between these systems significantly strengthens the antioxidant defense capacity of Termitomyces. This study is the first to elucidate the molecular network by which Asp enhances the antioxidant activity of Termitomyces, thereby providing a foundation for its development as a natural antioxidant. Full article

As a kind of eco-friendly material with wide application prospects, chitooligosaccharide (COS) has attracted increasing attention because of its unique bioactivities. In this study, novel polyphenol-functionalized COS pyridinium salts were designed and synthesized. The structural characteristics of the desired derivatives were confirmed by FT-IR and ¹H NMR spectroscopy. Their antioxidant activities were evaluated in vitro by DPPH radical scavenging assay, superoxide anion radical scavenging assay, and reducing power assay. The solubility assay in common solvents and cytotoxicity assay against L929 cells using the MTT method in vitro were also performed. The antioxidant assay results showed that the compounds functionalized by polyphenol displayed improved antioxidant activities, which were enhanced with the increase of sample concentration and the number of phenolic hydroxyl groups. The solubility assay indicated that the prepared derivatives had good water solubility. Besides, the modified products were non-toxic to the cells tested. In short, the polyphenol-functionalized COS pyridinium salts with enhanced antioxidant activity and good biocompatibility could be employed as newly safe antioxidant in the fields of biomedicine and food. Full article

Sustainable agriculture increasingly relies on biocontrol agents as eco-friendly solutions to combat plant diseases while improving soil health. In this context, species of the genus Bacillus, particularly Bacillus subtilis, have shown promise as effective biocontrol agents for plant diseases. This study demonstrates the successful fabrication of biohybrid materials by decorating electrospun poly(3-hydroxybutyrate) (PHB) fibers with electrosprayed chitooligosaccharide (COS) and Bacillus subtilis using simultaneous electrospinning and electrospraying. During electrospraying, COS formed a uniform film over the PHB fibers, serving as both an adhesive and a protective coating that maintained the viability and functionality of the embedded bacteria. SEM confirmed that bacterial spores were uniformly spread across the COS-coated biopolymer fibers. ATR-FTIR spectroscopy verified the successful deposition of COS on the fibers, while mechanical assay demonstrated enhancement in mechanical characteristics after the COS film formation on the PHB fibers compared to uncoated PHB. In addition, COS improved the wetting properties of the fibrous PHB material, creating a favorable environment for bacterial growth and development. Microbiological tests showed that the embedded B. subtilis remained viable and proliferated normally after 48 h under suitable conditions at 28 °C on agar medium. Furthermore, the biohybrid COS/B. subtilis-on-PHB materials also effectively inhibited the growth of pathogenic fungi, including species of Alternaria and Fusarium. These findings highlight the potential of dual electrospinning/electrospraying techniques for the fabrication of eco-friendly biocontrol formulations. The integration of COS coatings with B. subtilis provides a promising approach for sustainable agriculture by combining enhanced material properties with effective antifungal activity. Full article

Due to environmental pollution, unhealthy lifestyles, and autoimmunity, many individuals frequently encounter skin problems, such as sensitive skin, caused by the activation of the TRPV1 pathway and immune-inflammatory reactions. This study aims to synthesize compounds with soothing effects using β-chitooligosaccharides (COS) derived from marine Sargassum, which is investigated for its regulating inflammatory responses and activating of TRPV1 pathways. Dopamine is used as the hydroxyl donor in this process. COS-diDA, a compound that binds two dopamine amines, is synthesized using Michael additions and Schiff base reactions under alkaline circumstances, with a pH of 8.5. This work investigates the soothing effects of COS-diDA using a HaCaT cell model activated with capsaicin (CAP). ELISA and immunofluorescence experiments provide evidence that COS-diDA efficiently inhibits the activation of the TRPV1 protein in HaCaT cells by reducing its levels when stimulated with CAP. In addition, ELISA analysis demonstrates that COS-diDA reduces the secretion of IL-1α, IL-6, and IL-8 from HaCaT cells stimulated with CAP, indicating an interaction between the TRPV1 protein and interleukins. Additional research on in vitro pain models generated by CAP confirms the anti-inflammatory and soothing properties of COS-diDA. The study indicates that COS-diDA exhibits favorable adhesion qualities and possesses anti-inflammatory action, making it a viable candidate for inclusion as an anti-inflammatory component of cosmetic formulations. Full article

Flexible pressure sensors play an extremely important role in the fields of intelligent medical treatment, humanoid robots, and so on. However, the low sensitivity and the small initial capacitance still limit its application and development. At present, the method of constructing the microstructure of the dielectric layer is commonly used to improve the sensitivity of the sensor, but there are some problems, such as the complex process and inaccurate control of the microstructure. In this work, an ion composite photosensitive resin based on polyurethane acrylate and ionic liquids (ILs) was prepared. The high compatibility of the photosensitive resin and ILs was achieved by adding a chitooligosaccharide (COS) chain extender. The microstructure of the dielectric layer was optimized by digital light processing (DLP) 3D-printing. Due to the introduction of ILs to construct an electric double layer (EDL), the flexible pressure sensor exhibits a high sensitivity of 32.62 kPa⁻¹, which is 12.2 times higher than that without ILs. It also has a wide range of 100 kPa and a fast response time of 51 ms. It has a good pressure response under different pressures and can realize the demonstration application of human health. Full article

In this study, chitooligosaccharides (COS) were introduced into black soybean protein (BSP) using transglutaminase (TG) as a biocatalyst. The film-processing properties and physiological activities of the enzymatically glycosylated black soybean protein (EGBSP) were studied. The results showed that glycosylation decreased the surface hydrophobicity, absolute value of the zeta potential, its minimum solubility, and film permeability of BSP by 69.86%, 6.04%, 36.68%, and 14.91%, respectively, while increasing the tensile strength and elongation at break of its protein film by 56.57% and 172.68%, respectively. The gel time was shortened, and the acid-induced gel properties of EGBSP were similar to those of BSP. The anticancer effect of EGBSP was evaluated by the tumor inhibition rate, flow cytometry, and morphology observation of an ascites tumor in H₂₂ tumor-bearing mice. The immune organs (spleen, thymus), immune cells (lymphocytes, NK cells), and immune factors (IL-2, IL-12) of H₂₂ tumor-bearing mice were detected to evaluate the immunomodulatory effects of EGBSP. The results showed that medium and high doses of BSP had positive effects on immune enhancement and anti-cancer activity of H₂₂ tumor-bearing mice, while almost all doses of EGBSP showed significant effects. These results indicated that glycosylation significantly improved the anti-cancer effect and immunomodulatory activity of H₂₂ tumor-bearing mice while prolonging their overall survival. In conclusion, the glycosylation method using microbial transglutaminase to catalyze the introduction of chitooligosaccharides into black bean protein can improve the film-processing properties and biological activities of BSP more effectively than the enzyme crosslinking method. Full article

Chitooligosaccharides (COSs) are a class of functional carbohydrates with significant application prospects in food and medicine. Chitosanase CsnMY002 from the GH46 family has been used to prepare COS with controlled degrees of polymerization. To enhance the industrial applicability of CsnMY002, molecular dynamics (MD) simulations were applied to investigate the structure–property relationship. Guided by the simulation results, the beneficial mutants were screened through a synergistic strategy using a residue-folding free energy calculation and consensus sequence analysis. Iterative combinations constructed the mutant Mut6 (A49G/K70A/S84A/N89G/D199R/N221G) with significantly improved thermal stability, which had a half-life (t_1/2 value) at 55 °C and 75 °C that was 1.80 and 1.62 times higher than that of the wild type, respectively. A highly active mutant, Mut2, was created, exhibiting a 1.52 times catalytic efficiency of the wild type. An MD simulation analysis of the mutants suggested that the improved enzymatic properties were highly correlated with changes in the dynamic behaviours of the enzyme structure. This study generated more suitable CsnMY002 variants for COS production and provided a comprehensive strategy for the optimization of other industrial enzymes with application potential. Full article