Targeted metabolomics reveals the contribution of degradation and oxidation of lipids and proteins mediated by pH to the formation of characteristic volatiles in preserved egg yolk during pickling.

Targeted metabolomics and flavouromics combined with relative odor activity value were performed to explore the effect of degradation and oxidation of matrix mediated by pH on the formation of characteristic volatiles in preserved egg yolk (PEY) during pickling. It was found that the oxidation of proteins and lipids in PEY induced by pH sequentially occurred in early and later periods, and degradation both mainly occurred in early stage. Moreover, 1-octen-3-one, heptanal, trimethylamine, etc., compounds and 5-HETrE, proline, etc., components were confirmed as up-regulated characteristic volatiles and differential metabolites in PEY during pickling. The formation of octanal-M/D and benzeneacetaldehyde-M was attributed to ß-oxidation of hydroxyeicosapentaenoic acid and L-isoleucine catalyzed by strong alkali at early period based on correlation network between them, respectively. Meanwhile, the generation of 1-octen-3-one-M/D mainly depended on L-serine and could be promoted by phosphatidylcholines oxidation. At later stage, the formation of heptanal-M/D was primarily attributed to phosphatidylethanolamines oxidation induced by alkali, and the enrichment of heptanal-M/D and nonanal were both enhanced by oxidized lipids. Lastly, trimethylamine was derived from L-lysine under alkaline conditions and promoted by protein oxidation during the whole process. This manuscript provided insight into the differential contribution of oxidation and degradation from matrix regulated by exogenous factors on the formation pathway for characteristic volatiles in foods.

Formation mechanism and stability of egg white fluid gels under ultrahigh-pressure homogenization pretreatment and synergistic heating effect.

The thermal sterilization process of protein beverages inevitably leads to the formation of insoluble thermal aggregates, greatly reducing the texture and taste of protein beverages. In this study, homogenized egg white (HEW) was obtained by ultrahigh-high-pressure (UHP) homogenization pretreatment of egg white (EW), and then a special egg white fluid gel (EWFG) was prepared by water bath heating. The results showed that the optimal conditions for preparing EWFG were three cycles at 20 MPa homogenizing pressure and heating in a water bath at 72℃ for 10 min. Under these conditions, the EWFG was a milky-white homogeneous liquid with an average particle size of about 560 nm. Measurements of the physicochemical properties of HEW and EWFG showed that the UHP homogenization treatment reduced the viscosity of HEW, decreased the particle size of protein aggregates, and increased the zeta potential, which altered the interactions of proteins during the subsequent heating process and facilitated the formation of homogeneous and dispersed EWFG. EWFG showed good stability at pH 6-10 and in low-concentration saline and medium-concentration sucrose solutions. The EWFG obtained by the present treatment is more suitable for factory-scale production and has great potential for protein beverage applications.

Emulsifying properties of egg proteins: Influencing factors, modification techniques, and applications.

As an essential food ingredient with good nutritional and functional properties and health benefits, eggs are widely utilized in food formulations. In particular, egg proteins have good emulsification properties and can be commonly used in various food products, such as mayonnaise and baked goods. Egg protein particles can act as stabilizers for Pickering emulsions because they can effectively adsorb at the oil-water interface, reduce interfacial tension, and form a stable physical barrier. Due to their emulsifying properties, biocompatibility, controlled release capabilities, and ability to protect bioactive substances, egg proteins have become ideal carriers for encapsulating and delivering functional substances. The focus of this review is to summarize current advances in using egg proteins as emulsifiers. The effects of influencing factors (temperature, pH, and ionic strength) and various modification methods (physical, chemical, and biological modification) on the emulsifying properties of egg proteins are discussed. In addition, the application of egg proteins as emulsifiers in food products is presented. Through in-depth research on the emulsifying properties of egg proteins, the optimization of their applications in food, biomedical, and other fields can be achieved.

A new glycoprotein from pigeon egg: Study on its structure and digestive characteristics.

Pigeon egg white (PEW) is widely recognized as a promising source of bioactive proteins, with a high degree of glycosylation. This study focused on the characterization of a novel glycoprotein extracted from PEW, known as ovalbumin-related protein Y (OVAY). Our investigation included an analysis of the N-glycan and protein structures of OVAY, as well as an examination of simulated gastrointestinal digestive products and the transmembrane transport mechanism of OVAY-digested peptides. The results revealed that OVAY contains two glycosylation sites (Asn 62, 215) and consists of 30 N-linked glycoforms, with the top three glycans being N6H3, N6H7S1, and N6H5. Additionally, OVAY is rich in Gal and sialic acid and exhibits a rod-like molecular structure. Furthermore, it was found that OVAY demonstrates resistance to gastric digestion, with its digested peptides primarily transported via PepT1 and endocytosis. This study provides insight into the glycoprotein structure of OVAY and elucidates its physiological activity, providing a theoretical reference for the development of a novel sialate-rich protein.

Redistribution and fusion of protein-lipid assemblies within the egg yolk sphere under slight non-destructive deformation causing a change in thermal gel properties.

Egg yolk production processed after separating egg white is a common method to shorten cycle, but its taste quality will change even the vitelline membrane is intact. This might be related to the slight non-destructive deformation causing redistribution and fusion of protein-lipid assemblies within the egg yolk spheres. We investigated the mechanism of the change in thermal gel properties under slight deformation. The results of microscopic structural morphology revealed that the whole boiled egg yolk (WEY) underwent a transition in protein-lipid assembly morphology within yolk spheres, which changed from local aggregation to disordered fusion in shaken boiled egg yolks (SEYs). The spectroscopic and physicochemical properties analysis demonstrated that the redistribution of protein-lipid assemblies gave rise to marked changes in water migration, texture properties, molecular interactions, and oral sensation simulation of egg yolk thermal gels. This is benefit to guide the regulation of the taste quality egg yolk products in industry.

Insect-transmitted plant virus balances its vertical transmission through regulating Rab1-mediated receptor localization.

Rice stripe virus (RSV) establishes infection in the ovaries of its vector insect, Laodelphax striatellus. We demonstrate that RSV infection delays ovarian maturation by inhibiting membrane localization of the vitellogenin receptor (VgR), thereby reducing the vitellogenin (Vg) accumulation essential for egg development. We identify the host protein L. striatellus Rab1 protein (LsRab1), which directly interacts with RSV nucleocapsid protein (NP) within nurse cells. LsRab1 is required for VgR surface localization and ovarian Vg accumulation. RSV inhibits LsRab1 function through two mechanisms: NP binding LsRab1 prevents GTP binding, and NP binding LsRab1-GTP complexes stimulates GTP hydrolysis, forming an inactive LsRab1 form. Through this dual inhibition, RSV infection prevents LsRab1 from facilitating VgR trafficking to the cell membrane, leading to inefficient Vg uptake. The Vg-VgR pathway is present in most oviparous animals, and the mechanisms detailed here provide insights into the vertical transmission of other insect-transmitted viruses of medical and agricultural importance.

Chain reactions of temperature-induced egg white protein amorphous aggregates: Formation, structure and material composition of thermal gels.

Egg white protein is widely used in food, chemical, medical and other fields due to its excellent thermal gel properties. However, the regularity of egg white thermal gel (EWTG) by temperature influence is still unknown. In this study, we investigated the potential mechanism of temperature (75-95 °C, 15 min) gradient changes inducing thermal aggregation and gel formation of EWTG. The results showed that changes in textural characteristics and water holding capacity (WHC) of EWTGs depended on switching in protein aggregation morphology (spherical shape - chain shape - regiment shape) and gel network structure differences ("irregular bead-like" - "regular lamellar structure"). In addition, proteomics indicated that the generation of amorphous protein aggregates at 95 °C might be related to Mucin 5B as the aggregation core. The research revealed the EWTG formation from "whole egg white protein" to "single molecules", aiming to provide a reference for quality control in gel food processing.

Preparation of microgel particles from egg yolk components by combining phospholipase A2 with high-pressure homogenization: Physicochemical, structural properties and their effects on foaming, processing stability of egg white protein.

In this study, two types of microgel particles from egg yolk components were prepared by combining enzymatic hydrolysis with high-pressure homogenization (HPH), and their differences in physicochemical properties, foaming properties, and microstructure were compared. Results showed that the particle size of both types of microgel particles had decreased from 2744.07 ± 408.26 nm (egg yolk, EY) to 144.97 ± 3.19 nm (PLA2 hydrolyzed egg yolk microgel particles, PYM) and 535.07 ± 46.07 nm (egg yolk microgel particles hydrolyzed by PLA2, YMP), from 736.24 ± 34.61 nm (EG) to 182.76 ± 4.12 nm (PLA2 hydrolyzed egg yolk granules microgel particles, PGM) and 443.98 ± 27.09 nm (egg yolk granules microgel particles hydrolyzed by PLA2, GMP). Besides, their interfacial adsorption abilities were significantly improved, reflected in the increase values in overrun, from161.90 % ± 9.84 % (EY) to 269.64 % ± 16.73 % (PMY) and 307.20 % ± 16.09 % (YMP), from 189.21 % ± 5.02 % (EG) to 280.38 % ± 36.05 % (PGM) and 261.91 % ± 34.03 % (GMP). Their structural properties showed higher stabilities after treatments. When the microgel particles are applied to cakes, the specific volume was increased from 2.05 ± 0.1 mL/g (EY) to 2.25 ± 0.13 mL/g (PYM) and 2.45 ± 0.03 mL/g (YPM), and from 2.00 ± 0.09 mL/g (EG) to 2.51 ± 0.13 mL/g (PGM) and 2.75 ± 0.21 mL/g (GMP), respectively. The hardness and chewiness were reduced with both types of microgel particles from egg yolk components, which indicated their potential value as edible foam stabilizers in the baking industry.

Modification of the Protein Amino Acid Content in Hen Eggs as a Consequence of Different Concentrations of Lupine and Soy in Feed.

The effect of the diet modification (soybean and lupine addition) on the content of protein and amino acids (AA) in eggs was studied. Both the sampling day and the diet influenced the total protein content. In albumen, the lowest protein content (10.6%) was noted after administering a diet containing 25% lupine; in the same egg the yolk contained the most proteins (16.7%). In the content of nonessential AA (NAA) in egg yolks, differences were noted only for cysteine, with its the highest content in the yolks of the control group. The stable content of essential yolk amino acids (EAA) was observed only for isoleucine, leucine, tryptophan and phenylalanine. The highest contents of EAA and NAA were recorded in the yolks of the control group (~47 and ~53 g/100 g of protein, respectively) and in the group with 25% additions of lupine (~42 and ~51 g/100 g of protein, respectively). AA with constant content in the tested albumens were methionine, tryptophan and alanine. The highest content of EAA (>~42 g/100 g of protein) and NAA (>~62 g/100 g of protein) were determined in albumen of eggs determined in the group with at least 20% additions of lupine. The highest content of EAA for humans delivered eggs from groups 4-6 (with the addition of soy into the diet ≤5%). The protein sources used in the hen diet significantly influenced the content of protein and individual AA in the produced eggs.

Phylogenetic variations in a novel family of hyperstable apple snail egg proteins: insights into structural stability and functional trends.

The relationship between protein stability and functional evolution is little explored in proteins purified from natural sources. Here, we investigated a novel family of egg proteins (Perivitellin-1, PV1) from Pomacea snails. Their remarkable stability and clade-related functions in most derived clades (Canaliculata and Bridgesii) make them excellent candidates for exploring this issue. To that aim, we studied PV1 (PpaPV1) from the most basal lineage, Flagellata. PpaPV1 displays unparalleled structural and kinetic stability, surpassing PV1s from derived clades, ranking among the most hyperstable proteins documented in nature. Its spectral features contribute to a pale egg coloration, exhibiting a milder glycan binding lectin activity with a narrower specificity than PV1s from the closely related Bridgesii clade. These findings provide evidence for substantial structural and functional changes throughout the genus' PV1 evolution. We observed that structural and kinetic stability decreased in a clade-related fashion and was associated with large variations in defensive traits. For instance, pale PpaPV1 lectin turns potent in the Bridgesii clade, adversely affecting gut morphology, while giving rise to brightly colored PV1s providing eggs with a conspicuous, probably warning signal in the Canaliculata clade. This work provides a comprehensive comparative analysis of PV1s from various apple snail species within a phylogenetic framework, offering insights into the interplay among their structural features, stability profiles and functional roles. More broadly, our work provides one of the first examples from natural evolution showing the crucial link among protein structure, stability and evolution of new functions.

Impact of NaCl perturbation on physicochemical and structural properties of preheat-treated egg white protein modulating foaming property.

This study aimed to investigate the mechanism of NaCl perturbed preheat-treated egg white proteins' (EWPs) physicochemical and structural properties to modulate the foaming property (FP). The results revealed that NaCl regulated the salinolysis (5 mM) - salt precipitation (50 mM) - gradual or complete coverage with hydrated Na+ of the hydration layer (100-300 mM) - enhanced Cl- hydration repulsion (500 mM) of EWP, showing a gradual decrease in aggregates particle size, and reversibility of structural freedom, including moleculer flexibility and surface hydrophobicity. Whereas preheating temperature affected the secondary structure rearrangement and tertiary conformation exposure, and excessive temperature reduced foaming capacity while enhanced foam stability, with a tight correlation between NaCl-mediated EWPs' FP and the extent of Na+ covering the hydration layer. The findings provide a theoretical basis for processing factors to modulate the protein hydration layer to influence the functional properties.

Fabrication, characterization, and oxidation resistance of gelatin/egg white protein cryogel-templated oleogels through apple polyphenol crosslinking.

Cryogel-templated oleogels (CTO) were fabricated via a facile polyphenol crosslinking strategy, where apple polyphenol was utilized to crosslink the gelatin/egg white protein conjugates without forming hydrogels. After freeze-drying, cryogel templates were obtained and used to construct CTO by oil absorption. Apple polyphenol crosslinking improved the emulsion-related properties with appearance changes on samples, and infrared spectroscopy further confirmed the interactions between proteins and apple polyphenol. The crosslinked cryogels presented porous microstructures (porosity of over 96 %), enhanced thermal/mechanical stabilities, and could absorb a high content of oil (14.41 g/g) with a considerable oil holding capacity (90.98 %). Apple polyphenol crosslinking also influenced the rheological performances of CTO, where the highly crosslinked samples owned the best thixotropic recovery of 85.88 %. Moreover, after the rapid oxidation of oleogels, the generation of oxidation products was effectively inhibited by crosslinking (POV: 0.48 nmol/g, and TBARS: 0.53 mg/L). The polyphenol crosslinking strategy successfully involved egg white protein and gelatin to fabricate CTO with desired physical/chemical properties. Apple polyphenol acted as both a crosslinker and an antioxidant, which provided a good reference for fabricating pure protein-based CTO.

Insights from different reproductive gene knockdowns via RNA interference in the lady beetle Eriopis connexa: Establishing a new model for molecular studies on natural enemies.

Insect pest control can be achieved by the application of RNA interference (RNAi), a key molecular tool in functional genomics. Whereas most RNAi research has focused on insect pests, few studies have been performed on natural enemies. Validating the efficacy of RNAi in natural enemies is crucial for assessing its safety and enabling molecular research on these organisms. Here, we assessed the efficacy of RNAi in the ladybird beetle Eriopis connexa Germar (Coleoptera: Coccinellidae), focusing on genes related to reproduction, such as vitellogenin (Vg) and its receptor (VgR). In the transcriptome of E. connexa, we found one VgR (EcVgR) and two Vg genes (EcVg1 and EcVg2). These genes have been validated by in silico analyses of functional domains and evolutionary relationships. Five-day-old females were injected with 500 ng/µL of a specific double-stranded RNA (dsRNA) (dsEcVg1, dsEcVg2, or dsEcVgR) for RNAi tests, while nonspecific dsRNA (dsGFP or dsAgCE8.1) was used as a control. Interestingly, dsEcVg2 was able to knockdown both Vg genes, while dsEcVg1 could silence only EcVg1. Additionally, the viability of the eggs was significantly reduced when both Vg genes were knocked down at the same time (after treatment with dsEcVg2 or "dsEcVg1+dsEcVg2"). Ultimately, malformed, nonviable eggs were produced when EcVgR was silenced. Interestingly, no dsRNA treatment had an impact on the quantity of eggs laid. Therefore, the feasibility of RNAi in E. connexa has been confirmed, suggesting that this coccinellid is an excellent Neotropical model for molecular research on natural enemies and for studying RNAi nontarget effects.

Quality improvement of frozen cooked noodles by protein addition.

This study investigated the impact of protein enrichment on the physicochemical, cooking, textural, and color properties of frozen cooked noodles (FCN) stored for 0-3 weeks at -18 °C. Incorporating casein, egg white protein, and soy protein into the noodles significantly increased moisture content, with casein-enriched noodles showing the highest initial moisture levels. The addition of proteins also led to increased ash content, indicating improved nutritional quality. Protein enrichment resulted in reduced cooking loss and enhanced water retention during cooking and frozen storage. Casein-enriched noodles exhibited the highest water absorption capacity and the most substantial enhancement in textural properties, maintaining cohesiveness, gumminess, and elasticity better than egg white protein and soy protein during storage. The results indicated that egg white protein promotes intermolecular interactions, leading to enhanced color stability over time. These findings suggest that enriching with the protein could be a viable approach to elevate the overall quality of FCN.

Implications of intracrystalline OC17 on the protection of lattice incorporated proteins.

Biogenic CaCO3 formation is regulated by crystallization proteins during crystal growth. Interactions of proteins with nascent mineral surfaces trigger proteins to be incorporated into the crystal lattice. As a result of incorporation, these intracrystalline proteins are protected in the lattice, an example of which is ancient eggshell proteins that have persisted in CaCO3 for thousands of years even under harsh environmental conditions. OC17 is an eggshell protein known to interact with CaCO3 during eggshell formation during which OC17 becomes incorporated into the lattice. Understanding protein incorporation into CaCO3 could offer insights into protein stability inside crystals. Here, we study the protection of OC17 in the CaCO3 lattice. Using thermogravimetric analysis we show that the effect of temperature on intracrystalline proteins of eggshells is negligible below 250 °C. Next, we show that lattice incorporation protects the OC17 structure despite a heat-treatment step that is shown to denature the protein. Because incorporated proteins need to be released from crystals, we verify metal chelation as a safe crystal dissolution method to avoid protein denaturation during reconstitution. Finally, we optimize the recombinant expression of OC17 which could allow engineering OC17 for engineered intracrystalline entrapment studies.

Integrating Computational and Experimental Methods to Identify Novel Sweet Peptides from Egg and Soy Proteins.

Sweetness in food delivers a delightful sensory experience, underscoring the crucial role of sweeteners in the food industry. However, the widespread use of sweeteners has sparked health concerns. This underscores the importance of developing and screening natural, health-conscious sweeteners. Our study represents a groundbreaking venture into the discovery of such sweeteners derived from egg and soy proteins. Employing virtual hydrolysis as a novel technique, our research entailed a comprehensive screening process that evaluated biological activity, solubility, and toxicity of the derived compounds. We harnessed cutting-edge machine learning methodologies, specifically the latest graph neural network models, for predicting the sweetness of molecules. Subsequent refinements were made through molecular docking screenings and molecular dynamics simulations. This meticulous research approach culminated in the identification of three promising sweet peptides: DCY(Asp-Cys-Tyr), GGR(Gly-Gly-Arg), and IGR(Ile-Gly-Arg). Their binding affinity with T1R2/T1R3 was lower than -15 kcal/mol. Using an electronic tongue, we verified the taste profiles of these peptides, with IGR emerging as the most favorable in terms of taste with a sweetness value of 19.29 and bitterness value of 1.71. This study not only reveals the potential of these natural peptides as healthier alternatives to traditional sweeteners in food applications but also demonstrates the successful synergy of computational predictions and experimental validations in the realm of flavor science.

Oral Synergism of Egg-White-Derived Peptides (EWDP) and Curcumin for Colitis Mitigation via Polysaccharide/Cyclodextrin Metal-Organic Framework-Based Assemblies.

Recently, oral deliverable strategies of multiple nutraceuticals for ulcerative colitis (UC) mitigation have attracted increasing attention. This study aimed to fabricate facile oral assemblies loaded with egg-white-derived peptides (EWDP) and curcumin based on carboxymethyl chitosan (CMCS) and an γ-cyclodextrin metal-organic framework (MOF). Herein, outer CMCS could coassemble with EWDP (both nutraceuticals and building blocks) into cobweb-like fibrils to promote bridging with inner MOF via coordinative noncovalent interactions (hydrogen bonding, hydrophobic interaction, and electrostatic interaction). Compared with conventional γ-cyclodextrin/MOF-based composites, the above coassembly could also endow the biocompatible assemblies with superior nanoscale colloidal properties, processing applicability (curcumin storage stability, bioaccessibility, and aqueous solubility), and bioactivity. Moreover, the oral synergism of EWDP and curcumin (initially nonsynergistic) for UC mitigation was achieved by alleviating inflammatory damage and gut microbiota imbalance. Overall, the novel assemblies could be a promising amplifier and platform to facilitate oral formulations of various nutraceuticals for food processing and UC relief.

Comparative proteomic analysis of the ovarian fluid and eggs of Siberian sturgeon.

BACKGROUND: Sturgeon species are living fossils that exhibit unique reproductive characteristics, and elucidation of the molecular processes governing the formation and quality of sturgeon eggs is crucial. However, comprehensive data on the protein composition of sturgeon ovarian fluid (OF) and eggs and their functional significance are lacking. To address this knowledge gap, the aim of the present study was to conduct a comprehensive comparative proteomic analysis of Siberian sturgeon OF and eggs using liquid chromatography-mass spectrometry (LC-MS/MS). RESULTS: A total of 617 proteins were identified in OF, and 565 proteins were identified in eggs. A total of 772 proteins showed differential abundance. Among the differentially abundant proteins, 365 were more abundant in OFs, while 407 were more abundant in eggs. We identified 339 proteins unique to OFs and 287 proteins specific to eggs, and further investigated the top 10 most abundant proteins in each. The functional annotation of the OF proteins highlighted their predominant association with immune system processes, including the complement and coagulation cascade, neutrophil and leukocyte-mediated immunity, cholesterol metabolism, and regulation of the actin cytoskeleton. Analysis of egg proteins revealed enrichment in metabolic pathways, such as oxidative phosphorylation and fatty acid metabolism, and protein ubiquitination and translation. OF-specific proteins included extracellular matrix and secretory vesicles, and eggs were enriched in proteins localized to mitochondria and ribosome components. CONCLUSIONS: This study presents the first comprehensive characterization of the protein composition of sturgeon OF and eggs and elucidates their distinct functional roles. These findings advance our understanding of sturgeon reproduction, OF-egg signaling and the origin of OF proteins. The mass spectrometry proteomics data have been deposited in the ProteomeXchange Consortium with the dataset identifier PXD044168 to ensure accessibility for further research.

Assessing the structural and foaming property changes in egg yolk proteins due to malondialdehyde: Experimental and molecular docking studies.

This study evaluated the effects of varying levels of malondialdehyde (MDA) on the structural and foaming properties of the egg yolk proteins (EYPs), and the interaction between them was explored by molecular docking. The results showed that oxidative modification due to MDA increased the carbonyl content of EYPs by 4.49 times. Simultaneously, the total sulfhydryl content was reduced by 21.47%, and the solubility of EYPs was significantly decreased (p < 0.05). Continuous oxidation disorders the previously ordered structure of EYPs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that some proteins underwent crosslinking and aggregation with increased MDA oxidation, aligning with changes in particle size and zeta-potential. Moderate oxidation (<1 mmol/L) enhanced the foaming capacity and foam stability of EYPs. Additionally, molecular docking results uncovered favorable interactions between MDA and specific EYPs, primarily through hydrogen bonding. This research offers valuable insights into managing the functional and quality changes of yolk products during processing.

Hydrolyzed egg yolk peptide prevented osteoporosis by regulating Wnt/ß-catenin signaling pathway in ovariectomized rats.

Hydrolyzed egg yolk peptide (YPEP) was shown to increase bone mineral density in ovariectomized rats. However, the underlying mechanism of YPEP on osteoporosis has not been explored. Recent studies have shown that Wnt/ß-catenin signaling pathway and gut microbiota may be involved in the regulation of bone metabolism and the progression of osteoporosis. The present study aimed to explore the preventive effect of the YPEP supplementation on osteoporosis in ovariectomized (OVX) rats and to verify whether YPEP can improve osteoporosis by regulating Wnt/ß-catenin signaling pathway and gut microbiota. The experiment included five groups: sham surgery group (SHAM), ovariectomy group (OVX), 17-ß estradiol group (E2: 25 µg /kg/d 17ß-estradiol), OVX with low-dose YPEP group (LYPEP: 10 mg /kg/d YPEP) and OVX with high-dose YPEP group (HYPEP: 40 mg /kg/d YPEP). In this study, all the bone samples used were femurs. Micro-CT analysis revealed improvements in both bone mineral density (BMD) and microstructure by YPEP treatment. The three-point mechanical bending test indicated an enhancement in the biomechanical properties of the YPEP groups. The serum levels of bone alkaline phosphatase (BALP), bone gla protein (BGP), calcium (Ca), and phosphorus (P) were markedly higher in the YPEP groups than in the OVX group. The LYPEP group had markedly lower levels of alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) and C-terminal telopeptide of type I collagen (CTX-I) than the OVX group. The YPEP groups had significantly higher protein levels of the Wnt3a, ß-catenin, LRP5, RUNX2 and OPG of the Wnt/ß-catenin signaling pathway compared with the OVX group. Compared to the OVX group, the ratio of OPG/RANKL was markedly higher in the LYPEP group. At the genus level, there was a significantly increase in relative abundance of Lachnospiraceae_NK4A136_group and a decrease in Escherichia_Shigella in YPEP groups, compared with the OVX group. However, in the correlation analysis, there was no correlation between these two bacteria and bone metabolism and microstructure indexes. These findings demonstrate that YPEP has the potential to improve osteoporosis, and the mechanism may be associated with its modulating effect on Wnt/ß-catenin signaling pathway.