Enhancement of immunity and disease resistance in Litopenaeus vannamei through injection of tyramine formulated with polyethylene glycol.

This study investigates the prolonged effect of immune disease resistance in Litopenaeus vannamei through the administration of tyramine (TA) formulated with polyethylene glycol (PEG). Facing the challenges of intensive farming, environmental stress, and global climate changes, innovative approaches to improve shrimp health are essential. The research focuses on the role of biogenic amines in stress response and immune regulation, demonstrating that TA, especially when combined with PEG, significantly prolongs immunity and resistance against Vibrio alginolyticus. The experimental design included administering TA, PEG, and TA-PEG, followed by evaluations of immunity, lactate and glucose levels, and immune-related gene expressions. Results showed notable prolonged effects in total hemocyte count, phenoloxidase activity, and phagocytic activity in the TA-PEG group, indicating enhanced immune activation period. Additionally, the expression of prophenoloxidase system-related genes was significantly upregulated in the TA-PEG group. Furthermore, the TA-PEG group exhibited a significantly higher survival rate in a susceptibility test against V. alginolyticus. The results of this study confirm that the combined use of PEG can effectively extend the immunostimulatory duration of TA.

Specific Substrate Activity of Lotus Root Polyphenol Oxidase: Insights from Gaussian-Accelerated Molecular Dynamics and Markov State Models.

Polyphenol oxidase (PPO) plays a key role in the enzymatic browning process, and this study employed Gaussian-accelerated molecular dynamics (GaMD) simulations to investigate the catalytic efficiency mechanisms of lotus root PPO with different substrates, including catechin, epicatechin, and chlorogenic acid, as well as the inhibitor oxalic acid. Key findings reveal significant conformational changes in PPO that correlate with its enzymatic activity. Upon substrate binding, the alpha-helix in the Q53-D63 region near the copper ion extends, likely stabilizing the active site and enhancing catalysis. In contrast, this helix is disrupted in the presence of the inhibitor, resulting in a decrease in enzymatic efficiency. Additionally, the F350-V378 region, which covers the substrate-binding site, forms an alpha-helix upon substrate binding, further stabilizing the substrate and promoting catalytic function. However, this alpha-helix does not form when the inhibitor is bound, destabilizing the binding site and contributing to inhibition. These findings offer new insights into the substrate-specific and inhibitor-induced structural dynamics of lotus root PPO, providing valuable information for enhancing food processing and preservation techniques.

JrPPO1/2 play distinct roles in regulating walnut fruit browning by different spatiotemporal expression and enzymatic characteristics.

Polyphenol oxidase (PPO) activity drives walnut fruit browning, but the roles of its only two-family genes, JrPPO1 and JrPPO2, remain unclear. This study explores the spatiotemporal expression and enzymatic characteristics of JrPPO1 and JrPPO2 in walnut. Treatment with the PPO activator CuSO4 and H2O2 accelerated fruit browning and up-regulated JrPPO1/2 expression, whereas treatment with the PPO inhibitor ascorbic acid delayed browning, down-regulating JrPPO1 and up-regulating JrPPO2 expression. Compared to mJrPPO1, mJrPPO2 can exhibited better enzyme activity at higher temperatures (47 °C) and in more acidic environments (pH 4.25). mJrPPO2 exhibited a higher substrate specificity over mJrPPO1, and the preferred substrates are catechol, chlorogenic acid, and epicatechin. Additionally, mJrPPO2 adapted better to low concentration of oxygen (as low as 1.0% O2) and slightly elevated CO2 levels compared to mJrPPO1. Subcellular localization and spatiotemporal expression patterns showed that JrPPO1 is only expressed in green tissues and located in chloroplasts, while JrPPO2 is also located in chloroplasts, partly associated with membranes, and is expressed in both green and non-green tissues. Silencing JrPPO1/2 with virus-induced gene silencing (VIGS) reduced fruit browning, maintained higher total phenols, and decreased MDA production. Notably, silencing JrPPO1 had a greater impact on browning than JrPPO2, indicating JrPPO1's greater contribution to PPO activity and fruit browning in walnut fruits. Consequently, JrPPO1 can be effectively regulated both at the molecular level and by manipulating environmental conditions, to achieve the objective of controlling fruit browning.

Variations in quality attributes of pulsed light-treated table grape juice during refrigerated storage (4°C) and ambient conditions (25°C).

Pulsed light (PL) pasteurization is being explored as a substitute for the conventional thermal pasteurization of juices in recent times due to better retention of nutrients and overall quality. However, the long-term stability of the PL-pasteurized juice must be investigated to promote its application by the industry. The effect of PL treatment (effective fluence of 1.15 J·cm-2) and thermal treatment (90°C for 60 s) on microbial quality, enzyme activity, bioactive compounds, sensory acceptance, and color profile of table grape juice during storage at 4 and 25°C was investigated in this study. The PL pasteurization enhanced the microbial shelf-life of the juice (<6 log10cfu·mL-1) from 5 to 35 days at 4°C. The PL and thermally-pasteurized juice demonstrated a shelf-life of only 10 days when stored at 25°C. The total soluble solids and titratable acidity did not alter significantly throughout the storage period. The peroxidase, polyphenol oxidase, and pectin methylesterase activities were below 10% for the PL and thermally-treated beverage when stored at 4°C. The sensory acceptability of the PL-pasteurized juice after 35 days of refrigerated storage (6.9 ± 0.3) was close to the untreated juice (7.2 ± 0.3) and greater than thermally-treated juice (6.2 ± 0.2). After the 35th day of storage at 4°C, PL-treated grape juice retained 55%, 12%, and 15.3% more phenolics, flavonoids, and antioxidant capacity, respectively, than the thermally-pasteurized juice. Hence, PL pasteurization can effectively prolong the shelf-life of table grape juice while achieving microbial and enzymatic stability, along with high sensory and nutritional appeal. PRACTICAL APPLICATION: Exploring non-thermal methods like pulsed light (PL) pasteurization as a substitute for conventional thermal methods is gaining recognition for its ability to retain nutrients and improve overall juice quality. However, the industry's adoption depends on understanding the shelf-stability of PL-pasteurized juice. This study specifically investigates the practical applications of PL treatment in comparison with conventional thermal treatment in enhancing microbial safety and enzymatic stability in table grape juice. The findings contribute insights into optimizing the shelf life of table grape juice and preserving its quality, supported by microbial, enzymatic, and sensory evaluations.

Rational Design, Synthesis, and Computational Investigation of Dihydropyridine [2,3-d] Pyrimidines as Polyphenol Oxidase Inhibitors with Improved Potency.

Polyphenol oxidase (PPO) is an industrially important enzyme associated with browning reactions. In the present study, a set of ten new dihydropyridine [2,3-d] pyrimidines (TD-Hid-1-10) were synthesized and was found to be proven characteristically by 1H NMR, 13C NMR, IR, elemental analysis, and assessed as possible PPO inhibitors. PPO was purified from banana using three-phase partitioning, achieving an 18.65-fold purification and 136.47% activity recovery. Enzyme kinetics revealed that the compounds TD-Hid-6 and TD-Hid-7 are to be the most potent inhibitors, exhibiting mixed-type inhibition profile with IC50 values of 1.14 µM, 5.29 µM respectively against purified PPO enzyme. Electronic structure calculations at the B3LYP/PBE0 level of theories using def-2 SVP, def2-TZVP basis sets with various molecular descriptors characterized the electronic behavior of studied derivatives TD-Hid-1-10. Molecular electrostatic potential (MEP) and reduced density gradient analyses of RDG-NCI provided insights into charge distributions and weak intermolecular interactions. Docking study simulations predicted binding poses within crucial amino acid sequence in the 2y9x enzyme's active site, which is typically similar in sequence to the PPO form is not allowed. Ligands were analysed in terms of binding energies, inhibitor concentrations (mM) and various molecular interactions such as H-bonds, H-carbon, π-carbon, π-sigma, π-sigma, π-π T-shaped, π-π stacked, π-alkyl, Van der Waals and Cu interactions. The lowest binding energy (-7.83 kcal/mol) and the highest inhibitory effect (1.83 mM) were shown by the ligand Td-Hid-6, which forms H-bonds with Met280 and Asn260, exhibits π-sigma interactions with His61 and π-alkyl interactions with Val283. Other ligands also showed different interactions with various amino acids; for example, the Td-Hid-1 ligand formed H-bonds with His244 and showed π-sigma interactions with His244 and Val283.

Morphology of Four Strains of Phellinoid Agaricomycetes and Microstructural and Physiological Properties of Their Exudates.

To study and compare the morphology of the phellinoid Agaricomycetes strains and find other strategies to improve Phellinus spp. growth and metabolism. In this study, the morphological characteristics of four Phellinus igniarius strains (phellinoid Agaricomycetes) were observed under a light microscope. The exudates from these fungi were observed using light microscopy, scanning electron microscopy (SEM), and energy-dispersive spectrometry (EDS). The exudates were initially transparent with a water-like appearance, and became darker with time at neutral pH. Microscopy of air-dried exudates revealed regular shapes and crystals. Cl- (chloride) and K+ were the two key elements analyzed using EDS. Polyphenol oxidase (POD), catalase (CAT), and laccase activities were detected in mycelia from each of the four Phellinus strains. The K+ content of the three strains was higher than that of the wild strain. Cl- content correlated negatively with that of K+. Laccase activities associated with each mycelia and its corresponding media differed under cold and contaminated conditions.

Modified Metal-Organic Framework (MOF-818) inspired by natural enzymes for intelligent detection of total antioxidants and bisphenol A in infant beverages.

BACKGROUND: Accurate and quick judgement of the food quality can protect the legitimate rights of consumers. Currently, nanozymes are widely employed in the rapid detection of food due to their stability and economy. The contents of bisphenol A and antioxidant can be used to measure the quality of beverages. However, due to the complexity of the actual samples, it is still challenging to achieve the sensitive detection of both at the same time. The development of nanozyme with high enzyme activity is essential for sensitive detection of targets in complex foods. RESULTS: In this work, a novel nanomaterial (ZrTGA) was synthesized based on thioglycolic acid-modified Metal-Organic Framework (MOF-818). The interaction between Cu-S bonds and increase in the proportion of Cu1+ resulted in ZrTGA exhibiting higher peroxidase-like and polyphenol oxidase-like activities. These enzyme activities were 317 % and 200 % of the original values, respectively. With high enzyme activity can sensitively detect two important indicators of bisphenol A and antioxidants in beverages. The increased enzyme activity of ZrTGA enabled the content of both substances to be detected by smartphone extraction of RGB. Finally, through the output of the ''0″ and ''1″ signals of the logic gates, it is possible to quickly determine the level of the two substances and thus directly assess the quality of the beverages. SIGNIFICANCE: The modification of nanozyme enables the detection of substances at low concentrations based on enhancing dual-enzyme activity. The combination of mobile phone photography and logic gate technology enables the continuous detection of two important indicators in beverages, overcoming the limitations of traditional large-scale instruments. It also provides an alternative strategy for food quality detection.

Transcription factor MdbZIP44 targets the promoter of MdPPO2 to regulate browning in Malus domestica Borkh.

Apple (Malus domestica Borkh.) is among the most widely planted and economically valuable horticultural crops globally. Over time, the apple fruit's cut surface undergoes browning, and the degree of browning varies among different apple varieties. Browning not only affects the appearance of fruits but also adversely affects their taste and flavor. In the present study, we observed browning in different apple varieties over time and analyzed the expression of genes in the polyphenol oxidase gene family. The results indicated a strong correlation between the browning degree of the fruit and the relative expression of the polyphenol oxidase gene MdPPO2. With the MdPPO2 promoter as bait, the basic leucine zipper (bZIP) transcription factor MdbZIP44 was identified using the yeast single-hybrid screening method. Further investigation revealed that the overexpression of MdbZIP44 in 'Orin' callus could enhance the expression of MdPPO2 and promote browning of the callus. However, knocking out MdbZIP44 resulted in a callus with no apparent browning phenotype. In addition, our results confirmed the interaction between MdbZIP44 and MdbZIP11. In conclusion, the results indicated that MdbZIP44 can induce apple fruit browning by activating the MdPPO2 promoter. The results provide a theoretical basis for further clarifying the browning mechanism of apple fruit.

Lack of signal peptide in insect prophenoloxidase to avoid glycosylation to damage the zymogen activity.

Insect prophenoloxidases (PPOs) are important immunity proteins for defending against the invading pathogens and parasites. As a Type-Ⅲ copper-containing proteins, unlike Homo sapiens tyrosinases, the insect PPOs and most bacterial tyrosinases contain no signal peptides for unknown reason, however they can still be released. To this end, we fused different signal peptides to Drosophila melanogaster PPOs for in vitro and in vivo expression, respectively. We demonstrate that an artificial signal peptide can help PPO secretion in vitro. The secreted PPO appeared larger than wild-type PPO on molecular weight sizes due to glycosylation when expressed in S2 cells. Two asparagine residues for potential glycosylation in PPO1 were identified when a signal peptide was fused. After purification, the glycosylated PPO1 lost zymogen activity. When PPO1 containing a signal peptide was over-expressed in Drosophila larvae, the glycosylation and secretion of PPO1 was detected in vivo. Unlike insect PPO, human tyrosinase needs a signal peptide for protein expression and maintaining enzyme activity. An artificial signal peptide fused to bacterial tyrosinase had no influence on the protein expression and enzyme activity. These Type-Ⅲ copper-containing proteins from different organisms may evolve to perform their specific functions. Intriguingly, our study revealed that the addition of calcium inhibits PPO secretion from the transiently cultured larval hindguts in vitro, indicating that the calcium concentration may regulate PPO secretion. Taken together, insect PPOs can maintain enzyme activities without any signal peptide.

Ultraviolet C irradiation enhances the resistance of grape against postharvest black rot (Aspergillus carbonarius) by regulating the synthesis of phenolic compounds.

UV-C irradiation can maintain fruit quality by inducing fruit disease resistance and reducing decay during storage. Grape (Vitis Vinifera L.) was exposed to 2.4 kJ m-2 UV-C irradiation then inoculated with Aspergillus carbonarius to investigate the changes in nutritional quality, defense related substances and enzyme activities. Postharvest UV-C irradiation can increased the levels of defense-related substances and enzyme activities, such as phenols, flavanols, lignin, proline, glutathione, phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), and ß-1,3-glucanase (GLU). In addition, Resveratrol plays an important role in grape resistance to A. carbonarius infection through further verification by gene expression levels, the transcription factors VvWRKY24 and VvMYB14 are highly correlated with the regulation of VvSTS gene expression. This study revealed the molecular mechanism of postharvest grape fruit response to UV-C irradiation and the defense mechanism against black rot, and provided a theoretical basis for postharvest grape storage and preservation technology.

Improving blueberry cold storage quality: the effect of preharvest hexanal application on chilling injuries and antioxidant defense mechanisms.

Blueberries are vulnerable to chilling injury (CI). This can lead to limited longevity when they are subjected to cold storage conditions. This study investigated the effectiveness of a preharvest spray containing 0.02% hexanal in reducing CI and improving the postharvest storage quality of 'Star' and 'Biloxi' blueberries. The blueberries were stored for a period of 5 weeks at 2 °C and in 90% relative humidity (RH). The findings revealed that the preharvest hexanal spraying of both cultivars delayed senescence by mitigating CI, as evidenced by the bolstering of the antioxidant defense system through increased superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), catalase (CAT), and phenylalanine ammonia lyase (PAL) enzyme activity. The treated fruit also maintained elevated levels of total phenol content (TPC), total flavonoids (TFC), and vitamin C, demonstrating enhanced free radical scavenging capacity (FRSC), while exhibiting reduced polyphenoloxidase (PPO) activity, and reduced malondialdehyde (MDA), and H2O2 content in comparison with the control group. The preharvest hexanal treatment also suppressed fruit softening by maintaining greater firmness and higher membrane stability index (MSI) scores, inhibiting the activity of polygalacturonase (PG), pectinmethylesterase (PME), xylanase, and α-amylase, and reducing microbial counts (MC) and incidence of decay (DI) in comparison with the control. Preharvest hexanal treatment also improved the overall storage quality by reducing weight loss, total soluble solids (TSS), pH, and the TSS/acid ratio, while increasing titratable acidity (TA) in comparison with the control during cold storage. The findings suggest that hexanal, as a preharvest application, delays senescence effectively and preserves overall quality by enhancing cold tolerance through antioxidant defense mechanisms in blueberry storage under cold conditions. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

γ-aminobutyric acid treatment inhibits browning and promotes storage quality by regulating reactive oxygen species and membrane lipid metabolism in fresh-cut stem lettuce.

The effects of γ-aminobutyric (GABA) on enzymatic browning, storage quality, membrane and reactive oxygen species (ROS) metabolism in fresh-cut stem lettuce were investigated. The results illustrated that GABA treatment delayed browning degree, polyphenol oxidase (PPO) activity and the expression of LsPPO. Meanwhile, higher chlorophyll and ascorbic acid contents were exhibited in GABA-treated stem lettuce, as well as the slower microbial propagation. Further investigation revealed that exogenous GABA application declined malondialdehyde content, electrolyte leakage and the enzyme activities of membrane metabolism, and the expression levels of related genes were also downregulated. In addition, GABA treatment scavenged ROS and strengthened the enzyme activities of ROS metabolism, as well as the expression levels of corresponding genes. Taken together, these findings implied that the repressed enzymatic browning and microbial propagation in GABA-treated stem lettuce were due to the inhibition of ROS accumulation, enhancement of membrane stability and increased resistance to oxidation.

Impressive promiscuous biomimetic models of ascorbate, amine, and catechol oxidases.

Copper metalloenzymes ascorbate oxidase (AOase), amine oxidase (AmOase), and catechol oxidase (COase) possess copper(II) sites of coordination, which are trimeric, homodimeric, and dimeric, respectively. Two newly mononuclear copper(II) complexes, namely, [Cu(L)(bpy)](ClO4) (1) and [Cu(L)(phen)](ClO4) (2) where HL = Schiff base, have been synthesized. UV-visible, EPR and single-crystal X-ray diffraction examinations were used to validate the geometry in solution and solid state. For complex 1, the metal exhibits a coordination sphere between square pyramidal and trigonal bipyramidal geometry (τ, 0.49). A positive CuII/I redox potential indicates a stable switching between CuII and CuI redox states. Despite the monomeric origin, both homogeneous catalysts (1 or 2) in MeOH were found to favor three distinct chemical transformations, namely, ascorbic acid (H2A) to dehydroascorbic acid (DA), benzylamine (Ph-CH2-NH2) to benzaldehyde (Ph-CHO), and 3,5-di-tert-butylcatechol (3,5-DTBC) to 3,5-di-tert-butylquinone (3,5-DTBQ) [kcat: AOase, 9.6 (1) or 2.0 × 106 h-1(2); AmOase, 13.4 (1) or 9.4 × 106 h-1 (2); COase, 2.0 (1) or 1.9 × 103 h-1 (2)]. They exhibit higher levels of AOase activity as indicated by their kcat values compared to the AOase enzyme. The kcat values for COase activity in buffer solution [5.93 (1) or 2.95 × 105 h-1 (2)] are one order lower than those of the enzymes. This is because of the labile nature of the coordinated donor, the flexibility of the ligand, the simplicity of the catalyst-substrate interaction, and the positive CuII/I redox potential. Interestingly, more efficient catalysis is promoted by 1 and 2 concerning that of other mono- and dicopper(II) complexes.

Fluorescence detection of valence speciation of Cr(III) based on the catechol oxidase mimic enzyme activity of CuSeNP nanozymes.

Copper selenide nanoparticles (CuSeNP) were synthesized using histidine, ethylenediamine, and sodium selenate as precursors by one-step microwave digestion methods. The as-prepared CuSeNPs exhibit excellent catechol oxidase mimic enzyme and catalase (CAT)-like activities. Dopamine (DA) can be oxidized to aminochrome with H2O2 by CuSeNPs, and the intermediate product aminochrome can further react with α-naphthol to yield a highly fluorescent derivative. It was confirmed that Cr(III) could adsorb on the surface of CuSeNPs and inhibit the production of semiquinone radicals in the reaction system, and the catalytic activity of CuSeNPs was inhibited. The detection mechanisms, kinetics, and catalytic properties of CuSeNPs were systematically investigated. As a result, a novel fluorescence method for the assay of Cr(III) was established. The feasibility of CuSeNP nanozyme in detecting speciation Cr(III) in food samples was explored with satisfactory results. It showed the obvious potential for developing effective and dependable fluorescent detection method for protecting food safety.

Unveiling the long-term dynamic effects: Biochar mediates bacterial communities to modulate the petroleum hydrocarbon degradation in oil-contaminated sediments.

Sediment, as the destination of marine pollutants, often bears much more serious petroleum pollution than water. Biochar is increasingly utilized for remediating organic pollutant-laden sediments, yet its long-term impacts on oil-contaminated sediment remain poorly understood. In this study, simulation experiments adding 2.5 wt% biochars (corn straw and wood chips biochar at different pyrolysis temperatures) were conducted. The effects on petroleum hydrocarbon attenuation, enzyme activities, and microbial community structure were systematically investigated. Results showed enhanced degradation of long-chain alkanes in certain biochar-treated groups. Biochar species and PAH characteristics together lead to the PAHs' attenuation, with low-temperature corn straw biochar facilitating the degradation of phenanthrene, fluorene, and chrysene. Initially, biochars reduced polyphenol oxidase activity but increased urease and dehydrogenase activities. However, there was a noticeable rise in polyphenol oxidase activity for a long time. Biochars influenced bacterial community succession and abundance, likely due to nutrient release stimulating microbial activity. The structural equations model (SEM) reveals that DON affected the enzyme activity by changing the microbial community and thus regulated the degradation of PAHs. These findings shed light on biochar's role in bacterial communities and petroleum hydrocarbon degradation over extended periods, potentially enhancing biochar-based remediation for petroleum-contaminated sediments.

A comprehensive Study of Juglone's Effect on Polyphenol Oxidase in Cucumber: In Vitro Experiments and Computational Docking and Dynamics Insights.

This study explores the impact of juglone on cucumber (Cucumis sativus cv. Beith Alpha), scrutinizing its effects on seed germination, growth, and the polyphenol oxidase (PPO) enzyme's activity and gene expression. Employing concentrations ranging from 0.01 to 0.5 mM, we found juglone's effects to be concentration-dependent. At lower concentrations (0.01 and 0.1 mM), juglone promoted root and shoot growth along with germination, whereas higher concentrations (0.25 and 0.5 mM) exerted inhibitory effects, delineating a threshold for its allelopathic influence. Notably, PPO activity surged, especially at 0.5 mM in roots, hinting at oxidative stress involvement. Real-time PCR unveiled that juglone modulates PPO gene expression in cotyledons, peaking at 0.1 mM and diminishing at elevated levels. Correlation analyses elucidated a positive link between juglone-induced root growth and cotyledon PPO gene expression but a negative correlation with heightened root enzyme activity. Additionally, germination percentage inversely correlated with root PPO activity, while PPO activities positively associated with dopa and catechol substrates in both roots and cotyledons. Molecular docking studies revealed juglone's selective interactions with PPO's B chain, suggesting regulatory impacts. Protein interaction assessments highlighted juglone's influence on amino acid metabolism, and molecular dynamics indicated juglone's stronger, more stable binding to PPO, inferring potential alterations in enzyme function and stability. Conclusively, our findings elucidate juglone's dose-dependent physiological and biochemical shifts in cucumber plants, offering insights into its role in plant growth, stress response, and metabolic modulation.

Chitosan stabilized lemon essential oil nanoemulsion controls black mold rot and maintains quality of table grapes.

Aspergillus carbonarius infection leads to black mold rot in table grapes, causes grape decay, reduces fruit quality and marketability, which produces significant economic losses. This study investigated the antifungal efficacy of chitosan-stabilized lemon essential oil nanoemulsion (LO-CNE) against A. carbonarius and black mold rot of table grapes. LO-CNE was prepared with a mean diameter of 130.01 ± 8.34 nm. LO-CNE exhibited superior antifungal activity, reduced spore germination and germ tube elongation, decreased the antioxidant enzyme activities in A. carbonarius; the minimal inhibitory concentration of LO-CNE was determined to be 30 mg/mL. LO-CNE reduced the occurrence of black mold rot by 63 % and lesion diameter by 56.78 % in table grapes compared to the control. At their peak activity level, the grapes treated with LO-CNE exhibited significantly enhanced antioxidant and defense-related enzyme activities. Specifically, polyphenol oxidase activity increased by 2.27-fold, peroxidase activity by 2.22-fold, superoxide dismutase activity by 0.68-fold, catalase activity by 1.61-fold, phenylalanine ammonia-lyase activity by 3.38-fold, and ascorbate peroxidase activity by 2.36-fold. The LO-CNE application reduced natural decay by 95 %, weight loss by 15 % compared to the control, and effectively maintained the quality parameters of table grapes. Therefore, LO-CNE can be considered an alternative disease-control agent for grape preservation.

Impact of dietary administration of Arthrospira platensis free-lipid biomass on growth performance, body composition, redox status, immune responses, and some related genes of pacific whiteleg shrimp, Litopenaeus vannamei.

The current study aimed to assess the influence of dietary inclusion of cyanobacterium Arthrospira platensis NIOF17/003 as a dry material and as a free-lipid biomass (FL) on the growth performance, body composition, redox status, immune responses, and gene expression of whiteleg shrimp, Litopenaeus vannamei postlarvae. L. vannamei were fed five different supplemented diets; the first group was fed on an un-supplemented diet as a negative control group (C-N), the second group was fed on a commercial diet supplemented with 2% of A. platensis complete biomass as a positive control group (C-P20), whereas, the three remaining groups were fed on a commercial diet supplemented with graded amounts of FL at 1%, 2%, and 3% (FL10, FL20, and FL30, respectively). The obtained results indicated that the diet containing 1% FL significantly increased the growth performance, efficiency of consumed feed, and survival percentage of L. vannamei compared to both C-N and C-P20 groups. As for the carcass analysis, diets containing A. platensis or its FL at higher levels significantly increased the protein, lipid, and ash content compared to the C-N group. Moreover, the shrimp group fed on C-P20 and FL10 gave significantly stimulated higher digestive enzyme activities compared with C-N. The shrimp fed C-P20 or FL exhibited higher innate immune responses and promoted their redox status profile. Also, the shrimp fed a low FL levels significantly upregulated the expression of both the peroxiredoxin (Prx) and prophenoloxidase (PPO1) genes than those receiving C-N. The current results recommended that dietary supplementation with 1% FL is the most effective treatment in promoting the performance and immunity of whiteleg shrimp.

Inactivation of polyphenol oxidase by low intensity DC field: Experiment and mechanism analysis via molecular dynamics simulation and molecular docking.

In this study, inactivation of mushroom polyphenol oxidase (PPO) by low intensity direct current (DC) electric field and its molecular mechanism were investigated. In the experiments under 3 V/cm, 5 V/cm, 7 V/cm and 9 V/cm electric fields, PPOs were all completely inactivated after different exposure times. Under 1 V/cm, a residual activity of 11.88 % remained. The inactivation kinetics confirms to Weibull model. Under 1-7 V/cm, n value closes to a constant about 1.3. The structural analysis of PPO under 3 V/cm and 5 V/cm by fluorescence emission spectroscopy and molecular dynamics (MD) simulation showed that the tertiary structure was slightly changed with increased radius of gyration, higher potential energy and rate of C-alpha fluctuation. After exposure to the electric field, most of the hydrophobic tryptophan (TRP) residues turned to the hydrophilic surface, resulting the fluorescence red-shifted and quenched. Molecular docking indicated that the receptor binding domain of catechol in PPO was changed. PPO under electric field was MD simulated the first time, revealing the changing mechanism of the electric field itself on PPO, a binuclear copper enzyme, which has a metallic center. All these suggest that the low intensity DC electric field would be a promising option for enzymatic browning inhibition or even enzyme activity inactivation.

The role of a novel secretory peptidoglycan recognition protein with antibacterial ability from the Chinese Oak Silkworm Antheraea pernyi in humoral immunity.

Peptidoglycan recognition proteins (PGRPs) are a family of pattern recognition receptors that play a critical role in the immune response of invertebrates and vertebrates. Herein, the short ApPGRP-D gene was cloned from the model lepidopteran Antheraea pernyi. Quantitative PCR (qPCR) confirmed that ApPGRP-D is an immune-related protein and that the expression of ApPGRP-D can be induced by microorganisms. ApPGRP-D is a broad-spectrum pattern recognition protein that activates the prophenoloxidase cascade activation system and promotes the agglutination of microbial cells. Likely due to its amidase activity, ApPGRP-D can inhibit the growth of E. coli and S. aureus. In addition, we demonstrated for the first time that zinc ions, as important metal coenzymes, could promote multiple functions of ApPGRP-D but not its amidase activity.