Structural Basis of the Inhibition of L-Methionine γ-Lyase from Fusobacterium nucleatum.

Fusobacterium nucleatum is a lesion-associated obligate anaerobic pathogen of destructive periodontal disease; it is also implicated in the progression and severity of colorectal cancer. Four genes (FN0625, FN1055, FN1220, and FN1419) of F. nucleatum are involved in producing hydrogen sulfide (H2S), which plays an essential role against oxidative stress. The molecular functions of Fn1419 are known, but their mechanisms remain unclear. We determined the crystal structure of Fn1419 at 2.5 Å, showing the unique conformation of the PLP-binding site when compared with L-methionine γ-lyase (MGL) proteins. Inhibitor screening for Fn1419 with L-cysteine showed that two natural compounds, gallic acid and dihydromyricetin, selectively inhibit the H2S production of Fn1419. The chemicals of gallic acid, dihydromyricetin, and its analogs containing trihydroxybenzene, were potentially responsible for the enzyme-inhibiting activity on Fn1419. Molecular docking and mutational analyses suggested that Gly112, Pro159, Val337, and Arg373 are involved in gallic acid binding and positioned close to the substrate and pyridoxal-5'-phosphate-binding site. Gallic acid has little effect on the other H2S-producing enzymes (Fn1220 and Fn1055). Overall, we proposed a molecular mechanism underlying the action of Fn1419 from F. nucleatum and found a new lead compound for inhibitor development.

Structural and Biochemical Analyses of the Butanol Dehydrogenase from Fusobacterium nucleatum.

Butanol dehydrogenase (BDH) plays a significant role in the biosynthesis of butanol in bacteria by catalyzing butanal conversion to butanol at the expense of the NAD(P)H cofactor. BDH is an attractive enzyme for industrial application in butanol production; however, its molecular function remains largely uncharacterized. In this study, we found that Fusobacterium nucleatum YqdH (FnYqdH) converts aldehyde into alcohol by utilizing NAD(P)H, with broad substrate specificity toward aldehydes but not alcohols. An in vitro metal ion substitution experiment showed that FnYqdH has higher enzyme activity in the presence of Co2+. Crystal structures of FnYqdH, in its apo and complexed forms (with NAD and Co2+), were determined at 1.98 and 2.72 Å resolution, respectively. The crystal structure of apo- and cofactor-binding states of FnYqdH showed an open conformation between the nucleotide binding and catalytic domain. Key residues involved in the catalytic and cofactor-binding sites of FnYqdH were identified by mutagenesis and microscale thermophoresis assays. The structural conformation and preferred optimal metal ion of FnYqdH differed from that of TmBDH (homolog protein of FnYqdH). Overall, we proposed an alternative model for putative proton relay in FnYqdH, thereby providing better insight into the molecular function of BDH.

Structural and Functional Analysis of the Pyridoxal Phosphate Homeostasis Protein YggS from Fusobacterium nucleatum.

Pyridoxal 5'-phosphate (PLP) is the active form of vitamin B6, but it is highly reactive and poisonous in its free form. YggS is a PLP-binding protein found in bacteria and humans that mediates PLP homeostasis by delivering PLP to target enzymes or by performing a protective function. Several biochemical and structural studies of YggS have been reported, but the mechanism by which YggS recognizes PLP has not been fully elucidated. Here, we report a functional and structural analysis of YggS from Fusobacterium nucleatum (FnYggS). The PLP molecule could bind to native FnYggS, but no PLP binding was observed for selenomethionine (SeMet)-derivatized FnYggS. The crystal structure of FnYggS showed a type III TIM barrel fold, exhibiting structural homology with several other PLP-dependent enzymes. Although FnYggS exhibited low (<35%) amino acid sequence similarity with previously studied YggS proteins, its overall structure and PLP-binding site were highly conserved. In the PLP-binding site of FnYggS, the sulfate ion was coordinated by the conserved residues Ser201, Gly218, and Thr219, which were positioned to provide the binding moiety for the phosphate group of PLP. The mutagenesis study showed that the conserved Ser201 residue in FnYggS was the key residue for PLP binding. These results will expand the knowledge of the molecular properties and function of the YggS family.

Milk proteins and their derived peptides on bone health: Biological functions, mechanisms, and prospects.

Bone is a dynamic organ under constant metabolism (or remodeling), where a delicate balance between bone resorption and bone formation is maintained. Disruption of this coordinated bone remodeling results in bone diseases, such as osteoporosis, the most common bone disorder characterized by decreased bone mineral density and microarchitectural deterioration. Epidemiological and clinical evidence support that consumption of dairy products is beneficial for bone health; this benefit is often attributed to the presence of calcium, the physiological contributions of milk proteins on bone metabolism, however, are underestimated. Emerging evidence highlighted that not only milk proteins (including individual milk proteins) but also their derived peptides positively regulate bone remodeling and attenuate bone loss, via the regulation of cellular markers and signaling of osteoblasts and osteoclasts. This article aims to review current knowledge about the roles of milk proteins, with an emphasis on individual milk proteins, bioactive peptides derived from milk proteins, and effect of milk processing in particular fermentation, on bone metabolism, to highlight the potential uses of milk proteins in the prevention and treatment of osteoporosis, and, to discuss the knowledge gap and to recommend future research directions.

Preparation, Bioavailability, and Mechanism of Emerging Activities of Ile-Pro-Pro and Val-Pro-Pro.

Ile-Pro-Pro and Val-Pro-Pro are two most well-known food-derived bioactive peptides, initially identified as inhibitors of angiotensin I-converting enzyme (ACE) from a sample of sour milk. These two peptides were identified in fermented and enzymatic hydrolyzed cow and non-cow (that is, goat, sheep, buffalo, yak, camel, mare, and donkey) milk, as well as sourdough prepared from wheat, rye, and malt. Similar to other bioactive peptides, bioavailability of these peptides is low (about 0.1%), reaching picomolar concentration in human plasma; they showed blood pressure lowering activity in animals and in human, via improved endothelial function, activation of ACE2, and anti-inflammatory property. Emerging bioactivities of these two peptides toward against metabolic syndrome and bone-protection received limited attention, but may open up new applications of these peptides as functional food ingredients. Further studies are warranted to determine the best source as well as to identify novel enzymes (particularly from traditional fermented milk products) to improve the efficiency of production, to characterize possible peptide receptors using a combination of omics technology with molecular methods to understand if these two peptides act as signal-like molecules, to improve their bioavailability, and to explore new applications based on emerging bioactivities.

Structural and Biochemical Analysis of the Citrate-Responsive Mechanism of the Regulatory Domain of Catabolite Control Protein E from Staphylococcus aureus.

Catabolite control protein E (CcpE) is a LysR-type transcriptional regulator that positively regulates the transcription of the first two enzymes of the TCA cycle, namely, citZ and citB, by sensing accumulated intracellular citrate. CcpE comprises an N-terminal DNA-binding domain and a C-terminal regulatory domain (RD) and senses citrate with conserved arginine residues in the RD. Although the crystal structure of the apo SaCcpE-RD has been reported, the citrate-responsive and DNA-binding mechanisms by which CcpE regulates TCA activity remain unclear. Here, we report the crystal structure of the apo and citrate-bound SaCcpE-RDs. The SaCcpE-RD exhibits conformational changes between the two subdomains via hinge motion of the central ß4 and ß10 strands. The citrate molecule is located in a positively charged cavity between the two subdomains and interacts with the highly conserved Ser98, Leu100, Arg145, and Arg256 residues. Compared with that of the apo SaCcpE-RD, the distance between the two subdomains of the citrate-bound SaCcpE-RD is more than ∼3 Å due to the binding of the citrate molecule, and this form exhibits a closed structure. The SaCcpE-RD exhibits various citrate-binding-independent conformational changes at the contacting interface. The SaCcpE-RD prefers the dimeric state in solution, whereas the SaCcpE-FL prefers the tetrameric state. Our results provide insight into the molecular function of SaCcpE.

Crystal structure of the nicotinamidase/pyrazinamidase PncA from Bacillus subtilis.

The nicotinamidase/pyrazinamidase PncA is a member of a large family of hydrolase enzymes that catalyze the deamination of nicotinamide to nicotinic acid. PncA also functions as a pyrazinamidase in a wide variety of eubacteria and is an essential coenzyme in many cellular redox reactions in living systems. We report the crystal structure of substrate-free PncA from Bacillus subtilis (BsPncA) at 2.0 Šresolution to improve our understanding of the PncA family. The structure of BsPncA consists of an α/ß domain and a subdomain. The subdomain of BsPncA has a different conformation than that of PncA enzymes from other organisms. The B-factor analysis revealed a rigid structure of the α/ß domain, while the subdomain is highly flexible. Both dimers and tetramers were observed in BsPncA protein crystals, but only dimers were observed in solution. Our results provide useful information that will further enhance our understanding of the molecular functions of PncA family members.

[Correction Method of Atmospheric Scattering Effect Based on Three Spectrum Bands].

As a major error of CO2 retrieval, atmospheric scattering effect hampers the application of satellite products. Effect of aerosol and combined effect of aerosol and ground surface are important source of atmospheric scattering, so it needs comprehensive consideration of scattering effect from aerosol and ground surface. Based on the continuum, strong and weak absorption part of three spectrum bands O2-A, CO2 1.6 µm and 2.06 µm, information of aerosol and albedo was analyzed, and improved full physics retrieval method was proposed, which can retrieve aerosol and albedo simultaneously to correct the scattering effect. Simulation study on CO2 error caused by aerosol and ground surface albedo CO2 error by correction method was carried out. CO2 error caused by aerosol optical depth and ground surface albedo can reach up to 8%, and CO2 error caused by different types of aerosol can reach up to 10%, while these two types of error can be controlled within 1% and 2% separately by this correction method, which shows that the method can correct the scattering effect effectively. Through evaluation of the results, the potential of this method for high precision satellite data retrieval is obvious, meanwhile, some problems which need to be noticed in real application were also pointed out.

[Errors Analysis and Correction in Atmospheric Methane Retrieval Based on Greenhouse Gases Observing Satellite Data].

High precision retrieval of atmospheric CH4 is influenced by a variety of factors. The uncertainties of ground properties and atmospheric conditions are important factors, such as surface reflectance, temperature profile, humidity profile and pressure profile. Surface reflectance is affected by many factors so that it is difficult to get the precise value. The uncertainty of surface reflectance will cause large error to retrieval result. The uncertainties of temperature profile, humidity profile and pressure profile are also important sources of retrieval error and they will cause unavoidable systematic error. This error is hard to eliminate only using CH4 band. In this paper, ratio spectrometry method and CO2 band correction method are proposed to reduce the error caused by these factors. Ratio spectrometry method can decrease the effect of surface reflectance in CH4 retrieval by converting absolute radiance spectrometry into ratio spectrometry. CO2 band correction method converts column amounts of CH4 into column averaged mixing ratio by using CO2 1.61 µm band and it can correct the systematic error caused by temperature profile, humidity profile and pressure profile. The combination of these two correction methods will decrease the effect caused by surface reflectance, temperature profile, humidity profile and pressure profile at the same time and reduce the retrieval error. GOSAT data were used to retrieve atmospheric CH4 to test and validate the two correction methods. The results showed that CH4 column averaged mixing ratio retrieved after correction was close to GOSAT Level2 product and the retrieval precision was up to -0.24%. The studies suggest that the error of CH4 retrieval caused by the uncertainties of ground properties and atmospheric conditions can be significantly reduced and the retrieval precision can be highly improved by using ratio spectrometry method and CO2 hand correction method.

Development of chitosan-based edible film incorporated with purified flavonoids from Moringa oleifera: Structural, thermal, antibacterial activity and application.

Purified flavonoids (PF) from Moringa oleifera leaves were incorporated in chitosan (CS) polymer at different concentrations (0.5-4%) to produce a novel edible film. The physical, structure, mechanical, and bio-functional characterizations of the film were evaluated. The incorporation of PF significantly (p < 0.05) improved the thickness, solubility, swelling, and color of CS-films. Incorporating 4% of Moringa oleifera purified flavonoids (MOPF) improved the water vapor permeability from 8.85 to 2.47 g-1 s-1 Pa-1, and increased the film surface heterogeneity observed by SEM. Results also indicated that PF enhanced the mechanical properties and thermal stability of CS-films. The FTIR results indicated alterations in the CS-MOPF composite films' characteristics. Additionally, the incorporation of MOPF increased the antioxidation capacity. Furthermore, 4% of MOPF inhibited the activity of pathogenic bacteria in packed beef burgers. These results suggest that CS-MOPF composite films with enhanced technological and bio-functional properties could be industrially applied to increase the shelf-life of packaged foods.

Polysaccharides from Brassica rapa root: Extraction, purification, structural features, and biological activities. A review.

Brassica rapa (B. rapa) roots are attracting increased attention from nutritionists and health-conscious customers because of their remarkable performance in supplying necessary nutrients. Polysaccharides are major biologically active substances in B. rapa roots, which come in a variety of monosaccharides with different molar ratios and glycosidic bond types. Depending on the source, extraction, separation, and purification methods of B. rapa roots polysaccharides (BRP); different structural features, and pharmacological activities are elucidated. Polysaccharides from B. rapa roots possess a range of nutritional, biological, and health-enhancing characteristics, including anti-hypoxic, antifatigue, immunomodulatory, hypoglycemic, anti-tumor, and antioxidant activities. This paper reviewed extraction and purification methods, structural features, and biological activities as well as correlations between the structural and functional characteristics of polysaccharides from the B. rapa roots. Ultimately, this work will serve as useful reference for understanding the connections between polysaccharide structure and biological activity and developing novel BRP-based functional foods.

Seasonal and spatial distribution of 4-tert-octylphenol, 4-nonylphenol and bisphenol A in the Huangpu River and its tributaries, Shanghai, China.

The seasonal variations and spatial distributions of 4-tert-octylphenol (OP), 4-nonylphenol (NP) and bisphenol A (BPA) in surface waters, suspended solids and surface sediments in the Huangpu River and its tributaries (Suzhou River and Yunzao Brook) were firstly investigated. The mean concentrations of OP, NP and BPA in the three rivers were 10.59, 120.96 and 22.93 ng L(-1) in surface waters, 199.87, 2,300.87 and 84.11 ng g(-1) in suspended solids and 9.49, 119.44 and 7.13 ng g(-1) dry weight in surface sediments, respectively. The concentrations of NP and OP were higher in summer than in winter in the suspended solids and surface sediments, while the reverse was true in surface waters. Similarly, the levels of BPA were lower in summer than in winter in surface sediments, while the opposite was true in surface waters and suspended solids. These seasonal variations might be attributed to temperature and stream flows. High levels of OP, NP and BPA were found in surrounding river intersections, residential and industrial areas. Their concentrations decreased gradually with increasing distance from those areas, while the lowest levels were measured in near less urbanized and agricultural areas. These phenomena might indicate that the stream current and pollutant source were the major factors that affect the spatial distributions of OP, NP and BPA in the three rivers. Ecological risk assessment indicated that NP was the only one of the three pollutants with the potential to influence local aquatic organisms. The results of this study provide scientific support for control of these pollutants.

Whey Protein Hydrolysate Ameliorated High-Fat-Diet Induced Bone Loss via Suppressing Oxidative Stress and Regulating GSK-3ß/Nrf2 Signaling Pathway.

Long-term hypercaloric intake such as a high-fat diet (HFD) could act as negative regulators on bone remodeling, thereby inducing bone loss and bone microarchitecture destruction. Currently, food-derived natural compounds represent a promising strategy to attenuate HFD-induced bone loss. We previously prepared a whey protein hydrolysate (WPH) with osteogenic capacity. In this study, we continuously isolated and identified an osteogenic and antioxidant octapeptide TPEVDDA from WPH, which significantly promoted the alkaline phosphatase activities on MC3T3-E1 cells and exerted DPPH radical scavenging capacity. We then established an HFD-fed obese mice model with significantly imbalanced redox status and reduced bone mass and further evaluated the effects of different doses of WPH on ameliorating the HFD-induced bone loss and oxidative damages. Results showed that the administration of 2% and 4% WPH for 12 weeks significantly restored perirenal fat mass, improved serum lipid levels, reduced oxidative stress, and promoted the activity of antioxidant enzymes; meanwhile, WPH significantly preserved bone mass and bone mechanical properties, attenuated the degradation of trabecular microstructure, and regulated serum bone metabolism biomarkers. The protein levels of Runx2, Nrf2, and HO-1, as well as the phosphorylation level of GSK-3ß in tibias, were notably activated by WPH. Overall, we found that the potential mechanism of WPH on ameliorating the HFD-induced bone loss mainly through its antioxidant and osteogenic capacity by activating Runx2 and GSK-3ß/Nrf2 signaling pathway, demonstrating the potential of WPH to be used as a nutritional strategy for obesity and osteoporosis.

Casein Hydrolysate Alleviates Adipose Chronic Inflammation in High Fat-Diet Induced Obese C57BL/6J Mice through MAPK Pathway.

Obesity-induced adipose chronic inflammation is closely related to the development of insulin resistance and T2DM. Tripeptides l-valyl-l-prolyl-l-proline (VPP) and l-isoleucyl-l-prolyl-L-proline (IPP) derived from bovine casein have been reported to prevent inflammatory changes and mitigate insulin resistance in adipocytes. In this study, we aimed to investigate the influence of casein hydrolysates (CH) containing VPP and IPP on a high fat diet (HFD)-induced obese mice and cytokine TNF-α-induced adipocytes. Our data showed that CH alleviated chronic inflammation both in vivo and in vitro. 4% CH suppressed HFD-induced systemic inflammatory factors, hypertrophic white adipocytes, and macrophage infiltration. More importantly, CH was able to improve adipocyte dysfunction induced by TNF-α by increasing the expression of CCAAT/enhancer binding protein α (C/EBP-α) rather than peroxisome proliferator-activated receptor γ (PPAR-γ). Furthermore, CH also dose-dependently suppressed mitogen-activated protein kinase (MAPK)-c-Jun N-terminal kinase (JNK) phosphorylation and enhanced the phosphorylation of Erk 1/2, but not nuclear factor-kappa B (NF-κB) p65 phosphorylation, in TNF-α-induced 3T3-L1 cells. These results indicated that CH could ameliorate adipose chronic inflammation through the MAPK pathway. Altogether, our findings suggested that 4% CH supplementation for 6 weeks exerted a protective role in preventing obesity-related inflammation and adipose dysfunction.

Structural, in vitro digestion, and fermentation characteristics of lotus leaf flavonoids.

Bioaccessibility and bioactivity of flavonoids in lotus leaves are related to their characteristics in gastrointestinal digestion and colonic fermentation. The aim of this study is to investigate the stability of lotus leaf flavonoids (LLF) in simulated gastrointestinal digestion, and its modulation on gut microbiota in vitro fermentation. Results showed that LLF mainly consisted of quercetin-3-O-galactoside, quercetin-3-O-glucuronide, quercetin-3-O-glucoside, and kaempferol-3-O-glucoside. These flavonoids kept stability with only a small fraction degraded in simulated gastric and intestinal fluids. In vitro fermentation, LLF stimulated the growth of Actinobacteria and Firmicutes, inhibited the growth of Proteobacteria, and induced the production of fermentation gases and short-chain fatty acids. Interestingly, supplementation of soluble starch significantly improved the utilization of LLF by the intestinal flora. These results revealed that LLF shaped a unique biological web with Lactobacillus and Bifidobacterium spp. as the core of the biological network, which would be more beneficial to gut health.

[Expression, purification, and characterization of the histidine kinase CarS from Fusobacterium nucleatum].

Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 ß-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarScyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarScyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarScyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarScyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.

Digestion, absorption, and transport properties of soy-fermented douchi hypoglycemic peptides VY and SFLLR under simulated gastrointestinal digestion and Caco-2 cell monolayers.

Two novel hypoglycemic peptides VY and SFLLR were identified from douchi as the major peptides responsible for the glucose uptake activity. The present work aimed to elucidate their digestion, absorption and transport properties using simulated digestion and Caco-2 cell monolayers transport models. Besides, the effects of digestion and absorption on the structure and activity were also studied. The results showed that VY was resistant to gastrointestinal tract digestion and could cross Caco-2 cell monolayers intactly via both TJs-mediated passive paracellular pathway and PepT1-mediated active route. In comparison, SFLLR was partially degraded into small fragments of SFLL, SFL, and SF by the digestive system, leading to increased glucose uptake activity. Notably, SFLLR, SFLL, and SFL were partly hydrolyzed by aminopeptidase N or dipeptidyl peptidase IV during transport, but they were transported intact. SFL was transported via both paracellular diffusion and PepT1-mediated routes, while SFLLR and SFLL were via paracellular route only.

Douchi Peptides VY and SFLLR Improve Glucose Homeostasis and Gut Dysbacteriosis in High-Fat Diet-Induced Insulin Resistant Mice.

SCOPE: Two peptides VY (Val-Tyr) and SFLLR (Ser-Phe-Leu-Leu-Arg) are recently identified from soy-fermented douchi with hypoglycemic activity in cells. The study aims to understand their potential effects on glucose metabolism and insulin sensitivity as well as their mechanisms of action in a high-fat diet (HFD) induced insulin resistant model. METHODS AND RESULTS: C57BL/6 mice are fed HFD for 8 weeks, followed by peptide supplementation (doses: 10 and 50 mg kg-1 body weight) for 8 weeks. Peptides supplementation, especially SFLLR, reduces body weight gain, insulin resistance, hyperglycemia, inflammation, liver injury, and lipid accumulation. In both muscle and liver, both peptides activate glycogen synthase (GS), the key enzyme for glycogen synthesis, and also inhibit phosphoenolpyruvate carboxykinase (PEPCK) and glucose 6-phosphatase (G6PC), two rate-limiting enzymes for gluconeogenesis, via insulin and AMPK (5'-adenosine monophosphate-activated protein kinase) signaling pathways. Furthermore, VY and SFLLR supplementation reverse HFD-induced gut dysbacteriosis by decreasing the abundance of Enterococcus, Oscillibacter, and Deferribacter, and also increase the abundances of Alistipes, Lactobacillus, Faecalibaculum, Akkermansia, and Bifidobacterium (usually beneficial in the intestine). CONCLUSION: The study reveals the potential applications of peptides VY and SFLLR as a diet-based strategy for the prevention of type 2 diabetes.

Purification and characterization of hypoglycemic peptides from traditional Chinese soy-fermented douchi.

Douchi is a popular soy-fermented food that originated in China with documented hypoglycemic effects. However, the responsible molecules and the mechanism underlying their beneficial effects remain unclear. Therefore, in this study, we aimed to identify the responsible peptide(s) in douchi. A peptide extract of douchi was isolated step-wise by the C18 Sep-Pak technique, size exclusion chromatography, and semi-preparative liquid chromatography, and then the peptides were sequenced by UPLC-MS/MS. A total of 21 peptides were identified, of which three peptides, P3 (HPFR), P5 (VY), and P7 (SFLLR), were shown to improve glucose uptake in L6 cells. Both P5 and P7 increased glucose transporter 4 (GLUT4) translocation via the activation of AMPK and MAPK signaling pathways, but not the insulin-signaling pathway; adding an AMPK or an MAPK inhibitor prevented P5 or P7-induced glucose uptake as well as AMPK and MAPK activation. Our study showed that P5 and P7 could promote glucose uptake via AMPK and MAPK signaling pathways. In this study, two hypoglycemic peptides from douchi have been characterized for the first time.

Evaluation of the anti-osteoporotic effect of a low-phenylalanine whey protein hydrolysate in an ovariectomized mice model.

A phenylalanine (Phe)-restricted diet is indispensable to control the blood Phe for individuals with phenylketonuria (PKU), who are also confronted with progressive bone impairment. Thus, the development of a low-Phe protein substitute that could positively regulate bone metabolism is desired for their bone health. Our previous study reported the preparation of a low-Phe containing whey hydrolysate (LPH) from a selected whey protein hydrolysate (TAH). However, the effect of LPH on the bone status is unknown. In this study, we used an ovariectomized (OVX) mice model to evaluate the anti-osteoporotic potential of oral administration of whey protein concentrate (WPC, protein control), TAH, and LPH on bone physiology and bone metabolism. The results showed that after 12 weeks of treatment, the decreased bone mineral density, the deteriorated trabecular microarchitecture, and the reduced ultimate load due to ovariectomy were significantly attenuated by two whey protein hydrolysates (TAH and LPH); meanwhile, the body weight, uterine weight, bone composition, and the femoral elastic load of OVX mice had not been significantly affected by whey samples. In addition, LPH and TAH dual-regulated bone remodeling in OVX mice through triggering osteogenesis (promoted the expression of runt-related protein 2 (Runx2) and osteoformation markers) and inhibiting osteoresorption as well as inflammation. The modulated mitogen-activated protein kinase signaling and the inhibited nuclear factor κB signaling by LPH and TAH might relate to the dual-regulatory activities on bone. Overall, in the OVX mice model, LPH exerted higher osteoprotective potential than TAH of the same dose by activating the bone formation markers and inhibiting the inflammatory status. The current study demonstrated for the first time the potential use of a low-Phe whey hydrolysate, a protein substitute for PKU individuals, in the prevention of osteoporosis.