An inhibitor of the adaptability of Pseudomonas fluorescens in a high-salt environment. Phenomenon and mechanism of inhibition.

Pseudomonas fluorescens is a spoilage bacterium in food that has the ability to maintain growth and reproduction in high-salt environments. It acts as a defence mechanism through the exclusion of ions and the formation of biofilms. Hence, disrupting this defence mechanism may be a good way to control food spoilage. In this study, a specific flavonoid small molecule baicalin was found, which was able to dismantle the defence mechanism of the bacteria at a lower concentration (400 µM) of treatment. In synergy with salt, baicalin showed a significant inhibitory effect on the growth, c-di-gmp synthetics and biofilm formation of Pseudomonas fluorescens Pf08. Through transcriptomics, we also found that baicalein interfered with bacterial transport and polysaccharide production functions. Through molecular docking and QPCR, we found that baicalin is able to binding with the RpoS protein through hydrogen bonding and thus interfere with its function.

Design and characterization of an antibacterial film composited by hydroxyethyl cellulose (HEC), carboxymethyl chitosan (CMCS), and nano ZnO for food packaging.

For food packaging, a novel composite film was prepared by solution casting method using hydroxyethyl cellulose (HEC), carboxymethyl chitosan (CMCS), and zinc oxide nanoparticles as raw materials. The composite film successfully compounded the nanoparticles, as deduced by spectroscopy, crystallography and morphology observation. The addition of CMCS and ZnO enhanced the solvent resistance (the water solubility of the composite film was reduced by 94.3 %) and UV shielding ability (the UV shielding capacity of the composite film was increased by 45.73 %) of the composite film, thus improving the application prospects of the composite film in water-rich foods. In addition, the synergistic effect of CMCS and ZnO helped the composite film to efficiently inhibit the pathogenic bacteria Listeria monocytogenes and Pseudomonas aeruginosa (rate of inhibition>99.99 %) in food. The addition of CMCS and ZnO also significantly improved the elasticity (improve 494.34 %) and maximum load capacity (improve 142.24 %) of the composite film.

The nano antibacterial composite film carboxymethyl chitosan/gelatin/nano ZnO improves the mechanical strength of food packaging.

The carboxymethyl chitosan (CMCS)/fish skin gelatin (Gel) based novel nanocomposite film was developed with nano ZnO for potential food packaging applications. The SEM and FT-IR results indicated that the nano ZnO was success composited with CMCS/Gel film. The X-ray diffraction result revealed that the total crystallinity of the CMCS/Gel/nano ZnO achieved 94.92 %, improving the crystallinity of the original substrate. Compared with CMCS/nano ZnO and Gel/nano ZnO, the water solubility of CMCS/Gel/nano ZnO decreased to 23 %. Moreover, its contact angle reached 91°, representing that the composite film showed better solvent resistance and can be widely used in food packaging, especially in foods with high water content. After nano-ZnO was compounded with CMCS/Gel film, the physical properties were further improved. Furthermore, CMCS/Gel/nano ZnO has higher elasticity and ductility than CMCS/nano ZnO and Gel/nano ZnO. For food packages, CMCS/Gel films incorporated with nano ZnO depicted strong against Escherichia coli (99.20 %) and Staphylococcus aureus (84.70 %) for food packages. The CMCS/Gel film with the addition of ZnO was optimal for producing nanocomposite films with higher water-insolubility, elasticity and ductility, and higher antibacterial properties.

Exploring the interaction mechanism of dietary protein ovalbumin and folic acid: A combination research of molecular simulation technology and multispectroscopy.

This study aims to reveal the mechanism of the interaction between folic acid (FA) and egg ovalbumin (OVA) through the method of multi-spectroscopic, molecular docking, and molecular dynamics simulation in order to probe OVA as the possibility of a carrier of unstable vitamins. The results of the fluorescence spectra indicated a static quenching in the OVA-FA with a strong affinity of 6.998 × 104 M-1. At the same time, the complex formed by FA and OVA has changed the microenvironment. The measurement results of circular dichroism and particle size showed that FA and OVA gradually formed larger particles without changed the secondary structure of the protein. In addition, the results of molecular simulations indicated that the interaction between OVA and FA is mainly stabilized by strong hydrophobic and hydrogen bonds. This research was expanded the application prospect of dietary protein OVA as a transportation and protection system of vitamin substances.

Deciphering Changes in the Structure and IgE-Binding Ability of Ovalbumin Glycated by α-Dicarbonyl Compounds under Simulated Heating.

As a complex reaction, biological consequences of the Maillard reaction (MR) on dietary proteins need to be deciphered. Despite previous studies on the structural and antigenic properties of ovalbumin (OVA) by MR, associated changes induced by specific MR intermediates and their downstream products are largely unknown. This study focused on the impacts of glycation by α-dicarbonyl compounds (α-DCs), intermediates of MR and precursors of advanced glycation end-products (AGEs), on the structural and IgE-binding properties of ovalbumin (OVA) under simulated heating. Methylglyoxal (MGO), glyoxal (GO), and butanedione (BU) were selected as typical α-DCs to generate glycated OVA with different AGE-modifications (AGE-Ms). The results showed that reactions between OVA and α-DCs generated OVA-AGE with various degrees of modification and conformational unfolding, and the reactivity of α-DCs followed the order GO > MGO > BU. Depending on the precursor type, the levels of 10 specific AGEs were verified, and the amounts of total AGEs increased with heating temperature and α-DC dosage. Compared to native OVA, glycated OVA showed reduced IgE-binding levels but with sRAGE-binding ligands, the extent of which was associated with the contents of total AGEs and Nε-carboxymethyllysine, and changes in certain protein conformational structures. High-resolution mass spectrometry further identified different AGE-Ms on the Lys and Arg residues of OVA, confirming variations in the glycation sites and their associations with the immunoreactive epitopes of OVA under different conditions.

Ultrasonic pre-treatment modifies the pH-dependent molecular interactions between ß-lactoglobulin and dietary phenolics: Conformational structures and interfacial properties.

There is a need to understand the ultrasound-induced changes in the interactions between proteins and phenolic compounds at different pH. This study systematically explored the role of high-intensity ultrasound pre-treatment on the binding mechanisms of ß-lactoglobulin (ß-LG) to two common phenolic compounds, i.e., (-)-epigallocatechin-3-gallate (EGCG) and chlorogenic acid (CA) at neutral and acidic pH (pH 7.2 and 2.4). Tryptophan fluorescence revealed that compared to proteins sonicated at 20% and 50% amplitudes, 35%-amplitude ultrasound pre-treatment (ULG-35) strengthened the binding affinities of EGCG/CA to ß-LG without altering the main interaction force. After phenolic addition, ULG-35 displayed a similar but a greater extent of protein secondary and tertiary structural changes than the native protein, ascribed to the ultrasound-driven hydrophobic stacking among interacted molecules. The dominant form of ß-LG (dimer/monomer) played a crucial role in the conformational and interfacial properties of complexes, which can be explained by the distinct binding sites at different pH as unveiled by molecular docking. Combining pre-ultrasound with EGCG interaction notably increased the foaming and emulsifying properties of ß-LG, providing a feasible way for the modification of bovine whey proteins. These results shed light on the understanding of protein-phenolic non-covalent binding under ultrasound and help to develop complex systems with desired functionality and delivery.

Transcription factors FabR and FadR regulate cold adaptability and spoilage potential of Shewanella baltica.

Shewanella baltica is the specific spoilage microorganism of Pseudosciaena crocea during low-temperature storage. Exploring the correlation between cold adaptability and spoilage potential may provide a new perspective for prolonging shelf life of aquatic products. In the present study, we investigated the synthesis pathway of unsaturated fatty acid (UFA) responsible for regulating cold adaptability in Shewanella baltica and its effect on spoilage potential. FabR and FadR, as key regulators of membrane unsaturated fatty acids synthesis pathway, were identified in S. baltica. FabR was significantly down-regulated at 4 °C compared to at 30 °C, yet FadR displayed the opposite results. By overexpressing fabR and fadR genes at 4 °C, we found that FabR and FadR had negative and positive effects on UFA content and membrane fluidity as well as spoilage potential, respectively. These data indicated that FabR and FadR functioned collectively to increase the membrane fluidity for better cold adaptability at low temperature, resulting in the maintenance of spoilage potential of S. baltica.

MgO/carboxymethyl chitosan nanocomposite improves thermal stability, waterproof and antibacterial performance for food packaging.

A novel nanocomposite film was fabricated by carboxymethyl chitosan (CMCS) and nano MgO for potential food packaging applications. The impregnation of MgO nanoparticles into CMCS was evidenced by the X-ray diffraction and FTIR spectroscopy. SEM micrographs revealed a dense layer of MgO formation in the CMCS matrix, which is a major contributor to the improvement of crystallinity. Compared with pure CMCS, CMCS/MgO composites confer improved thermal stability, better UV shielding performance, as well as water-insolubility, improving the feasibility of using CMCS-based biopolymer films as food packagings, especially in the case of water-rich food. These physical properties were further enhanced with the increase in MgO content. Furthermore, MgO nanoparticles can simultaneously provide CMCS with increased elasticity and ductility at a rather low filler content (1.0 % by weight). For biological properties, CMCS/MgO composites exhibited excellent antimicrobial activity against Listeria monocytogenes and Shewanella baltica.

A tandem GGDEF-EAL domain protein-regulated c-di-GMP signal contributes to spoilage-related activities of Shewanella baltica OS155.

Cyclic diguanylate (c-di-GMP) is a second messenger involved in the regulation of various physiological processes in bacteria. However, its function in spoilage bacteria has not yet been addressed. Here, we studied the function of a tandem GGDEF-EAL domain protein, Sbal_3235, in the spoilage bacterium Shewanella baltica OS155. The deletion of sbal_3235 significantly reduced the c-di-GMP level, biofilm formation, and exopolysaccharide, trimethylamine (TMA), and putrescine production; sbal_3235 deletion also downregulated the expression of the torS and speF genes and affected membrane fatty acid composition. Site-directed mutagenesis in conserved GGDEF and EAL motifs abolished diguanylate cyclase (DGC) and phosphodiesterase (PDE) activity, respectively. These data indicate that Sbal_3235 is an essential contributor to the c-di-GMP pool with bifunctional DGC and PDE activity, which is involved in the biofilm formation and spoilage activity of S. baltica OS155. Our findings expand the biochemical role of c-di-GMP and uncover its link to spoilage activities, providing novel targets for food quality and safety controlling.