Effect of low-voltage electrostatic field-assisted partial freezing on large yellow croaker protein properties and metabolomic analysis during storage.

BACKGROUND: Large yellow croaker is highly perishable during storage because of high protein and moisture content. The degradation of the fish is mainly attributed to microbial growth and enzyme activity, so it is important to find an efficient storage method to extend its shelf life. METHODOLOGY: This study investigated the effect of a low-voltage electrostatic field combined with partial freezing treatment on the physicochemical properties of myofibrillar protein (MP) and metabolomic analysis of large yellow croaker during preservation. The samples in chilled storage (C), partial freezing storage (PF) and 6 kV/m low-voltage electrostatic field partial freezing storage (LVEF-PF) were analyzed during an 18 day storage period. RESULTS: In comparison with the C and PF groups, LVEF-PF delayed the oxidation of MP by inhibiting the formation of carbonyl groups (2.25 nmol/mg pro), and maintaining higher sulfhydryl content (29.73 nmol/mg pro). Fourier transform infrared (FTIR) spectroscopy and fluorescence spectroscopy analysis also demonstrated that the LVEF-PF treatment maintained the stability of the protein structure by increasing the a-helix ratio (19.88%) and reducing the random coil ratio (17.83%). Scanning electron microscopy showed that, compared with the LVEF-PF group, there was more degeneration and aggregation of MP in the C and PF groups after 18 days' storage. The results of untargeted metabolomic analysis showed that 415 kinds of differential metabolites were identified after storage, and the difference levels of differential metabolites were least between the samples treated with LVEF-PF stored on the ninth day and the fresh samples. The main differential metabolic pathways during storage were amino acid metabolism and lipid metabolism. CONCLUSION: The LVEF-PF treatment could maintain the stability of myofibrillar protein in large yellow croaker during storage. These results showed a potential application of the LVEF-PF method for aquatic product preservation. © 2023 Society of Chemical Industry.

Effect of low voltage electrostatic field combined with partial freezing on the quality and microbial community of large yellow croaker.

The effect of low voltage electrostatic field combined with partial freezing (LVEF- PF) treatment on storage quality and microbial community of large yellow croaker was studied. Three different methods including chilled (C), partial freezing (PF) and 6 kV/m electrostatic field combined partial freezing storage were used to preserve large yellow croaker for 18 days. Total viable counts (TVC), sensory evaluation, and physiochemical index including pH, total volatile basic nitrogen (TVB-N), K value and centrifugal loss were examined. During storage, the large yellow croaker was susceptible to microbial growth and spoilage. However, LVEF-PF treatment was found to be effective in enhancing sensory quality, inhibiting microbial growth, and maintaining myofibril microstructure. Low field nuclear magnetic resonance showed that LVEF-PF treatment reduced the migration of immobilized water to free water. At 18th day, the TVC value of LVEF-PF, PF and chilled group were 3.56 log CFU/g, 5.11 log CFU/g, 7.73 log CFU/g, respectively. Therefore, from the results of TVB-N and TVC value, the shelf life of LVEF-PF group was at least 3 days longer than PF group, and 6 days longer than the chilled group. High-throughput sequencing showed that the microbial community diversity significantly decreased during storage. The predominant bacteria in chilled, PF, LVEF-PF group at 18th day were Pseudomonas, Psychrobacter and Shewanella, respectively, and the relative abundance of spoilage bacteria such as Pseudomonas and Psychrobacter were reduced by LVEF-PF treatment, that corresponding with lower values of TVB-N and TVC value. LVEF-PF treatment could be used as a new processing and storage method to delay deterioration and prolong shelf life of large yellow croaker.

Study on Purification and Characterization of Polyphenol Oxidase from Acetes chinensis.

Acetes chinensis (belonging to the Decapoda Sergestidae genus) is widely distributed in East Asian waters and is extremely widespread and present in the shallow coastal areas of China. Polyphenol oxidase (PPO), which was extracted from Acetes chinensis, was purified in a four-step procedure involving phosphate-buffered saline treatment, ammonium sulphate precipitation, DEAE-Cellulose chromatography, and Phenyl-Sepharose HP chromatography, and then, its biochemical characterization was measured. The specific activity of the purified enzyme was increased to 643.4 U/mg, which is a 30.35 times increase in purification, and the recovery rate was 17.9%. L-dopa was used as the substrate, the enzymatic reactions catalyzed by PPO conformed to the Michaelis equation, the maximum reaction velocity was 769.23 U/mL, and the Michaelis constant Km was 0.846 mmol/L. The optimal pH of PPO from Acetes chinensis was 7.5, and the optimal temperature was 35 °C. The metal ions experiment showed that Mn2+ and K+ could enhance the activity of PPO; that Ba2+ and Ca2+ could inhibit the activity of PPO; and that Cu2+ had a double effect on PPO, increasing the PPO activity at low concentrations and inhibiting the PPO activity at high concentrations. The inhibitor experiment showed that the inhibitory effects of EDTA and kojic acid were weak and that ascorbic acid and sodium pyrophosphate had good inhibitory effects. The purification and characterization of Acetes chinensis serve as guidelines for the prediction of enzyme behavior, leading to effective prevention of enzymatic browning during processing.

Effect of infrared radiation on activity and conformation of polyphenol oxidase from Acetes chinensis.

The aim of this paper was to study the effect of infrared radiation (IR) on the activity and conformation of polyphenol oxidase (PPO) in Acetes chinensis. In this paper, the specific activity of PPO was increased from 21.2 to 643.4 U/mg by a four-step purification. The results showed that IR treatment had greater effect on the enzyme activity and conformation of PPO than hot air (HA) treatment. After IR treatment at 70°C, the relative enzyme activity of PPO was 9.28%, the surface hydrophobicity index increased by 80.42%, and the content of sulfhydryl group decreased to 96.99% of the control group. The results of circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR) showed that the α-helix of PPO treated by IR decreased and the random coil increased. The intrinsic fluorescence intensity of PPO decreased after IR treatment, indicating that the tertiary structure of PPO was destroyed. Scanning electron microscopy (SEM) results showed that the surface microstructure of PPO after IR treatment became clear and compact. In conclusion, IR treatment can completely destroy the secondary structure and tertiary structure of PPO and cause enzyme inactivation. This study provides a treatment for reducing the activity of PPO from A. chinensis during the production and processing. PRACTICAL APPLICATION: This study shows that IR treatment has a better inhibitory effect on the activity of PPO than HA treatment. It provides a better treatment method for inactivating the activity of PPO from Acetes chinensis during the production and processing.

microRNA-495 reduces visceral sensitivity in mice with diarrhea-predominant irritable bowel syndrome through suppression of the PI3K/AKT signaling pathway via PKIB.

Diarrhea-predominant irritable bowel syndrome (IBS-D) is one of the most common gastrointestinal disorders in the world, lacking effective therapies. The crucial roles of microRNAs (miRNAs) in IBS-D have attracted increasing attention. The aim of this study is to investigate the effects of miR-495 on the visceral sensitivity of the IBS-D through the PI3K/AKT signaling pathway by targeting PKIB. Microarray data analysis was employed to screen the differentially expressed genes related to IBS-D and regulatory miRNAs. Then, mice were perfused with acetic acid into the rectum to establish the IBS-D model. Next, PKIB expression was measured in IBS-D mice. Additionally, model mice were injected with a series of adenovirus vector to investigate the influence of miR-495 on visceral sensitivity and rectal function in IBS-D mice with the involvement of PKIB and PI3K/AKT signaling pathway. The IBS-D mouse model was successfully established. PKIB was the target gene of miR-495, and highly expressed in mice with IBS-D. Silencing PKIB reduced visceral sensitivity in mice with IBS-D, and overexpression of miR-495 decreased visceral sensitivity in mice with IBS-D by inhibiting PKIB. Moreover, miR-495 upregulation inhibited PI3K/AKT signaling pathway through downregulating PKIB. To sum up, this study reveals that miR-495 upregulation can reduce visceral sensitivity in IBS-D via inhibition of PI3K/AKT signaling pathway by targeting PKIB. It suggests that miR-495 presents a potential target for IBS-D therapy.

Shape-controlled synthesis of 3D and 1D structures of CdS in a binary solution with L-cysteine's assistance.

A facile L-cysteine-assisted route was designed for the selectively controlled synthesis of 1D and novel, interesting 3D CdS spherical nanostructures constructed from CdS nanorods (or nanopolypods) in a binary solution. By controlling reaction conditions such as the molar ratio between Cd(OAc)2 and L-cysteine and the volume ratio of the mixed solvents, the synthesis of various 3D architectural structures and 1D wirelike structures in large quantities can be controlled. This is the first reported case of the direct growth of novel 3D self-assemblies of CdS nanorods (or nanopolypods). The morphology, structure, and phase composition of the as-prepared CdS products were examined by using various techniques (X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), selected-area electron diffraction (SAED), high-resolution TEM, and Raman spectroscopy). On the basis of the results from TEM studies and our analysis, we speculate that in the present synthesis the L-cysteine dominates nucleation growth and the ethylenediamine (en)-dominated, oriented-assembly process. Interestingly, the products obtained show a gradient evolution in color from light-yellow to dark-yellow, which implies that their intrinsic optical properties change, possibly due to variations in their special morphologies and structures. This facile solution-phase L-cysteine-assisted method could be extended for the controlled preparation of other metal chalcogenides nanostructures with complex morphologies.