The effects of heat treatment(heating temperature and pH) on the structures and emulsifying properties of caseins were systematically studied by spectroscopy.Heat treatment from 60to 100℃resulted in an increase in th...The effects of heat treatment(heating temperature and pH) on the structures and emulsifying properties of caseins were systematically studied by spectroscopy.Heat treatment from 60to 100℃resulted in an increase in their fluorescence intensity,hydrodynamic diameter,turbidity and emulsifying activity index,but decreased the size polydispersity of caseins.In the pH range of 5.5to 7.0,the fluorescence intensity,hydrodynamic diameter,turbidity and emulsifying properties decreased with increased heating pH,but the size polydispersity of caseins increased with increased pH.The relationship between the surface fluorescence intensity and emulsifying activity was also investigated,revealing a correlation coefficient of 0.90.These results suggested that heat treatment could be used to modify the structures and emulsifying properties of caseins by appropriately selecting heating conditions.展开更多
Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and e...Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.展开更多
This paper carried out a comparative analysis of different types of electrophoretic systems which were used for the analysis of casein complex from cow milk (polyacrylamide gel electrophoresis: for the neutral and aci...This paper carried out a comparative analysis of different types of electrophoretic systems which were used for the analysis of casein complex from cow milk (polyacrylamide gel electrophoresis: for the neutral and acidic native conditions, in gradient variant, the presence of sodium dodecyl sulphate or with including urea). Taking into attention the separation efficiency, complexity of electrophoretic system, the impact of system components, we have selected the anode system of the homogeneous gel in the presence of urea as the basis for the preparation of casein fractions. It also changed the composition and the structure of the electrophoretic apparatus. The changes allow purification of casein fractions up to several grams during one stage of treatment (for 5 hours). The purified casein fractions were tested for the homogeneity and have been recommended for using in the biomedical researches, including the processes of the formation of the bioactive peptides.展开更多
Polyembryony has posed a significant impediment to the advancement of citrus hybrid breeding.FhRWP is widely regarded as a pivotal factor governing asexual reproduction in citrus,and prior research has demonstrated th...Polyembryony has posed a significant impediment to the advancement of citrus hybrid breeding.FhRWP is widely regarded as a pivotal factor governing asexual reproduction in citrus,and prior research has demonstrated that FhARID1,acting as an upstream regulator,modulates FhRWP expression.In this study,we performed a genome-wide characterization of the ARID-HMG-related genes using the short juvenile minicitrus Fortunella hindsii.A total of 20 ARID-HMG-related genes were identified.Protein interaction network and enrichment analysis suggested that ARID-HMG-related proteins might might be involved in chromatin remodeling complexes.Knockout of FhARID1 in F.hindsii did not induce the conversion from polyembryony to monoembryony.However,fharid1 plants in T1 generation exhibited abnormal proliferation at axillary buds,which is similar to phenotype of fhrwp plants.Expression analysis of fharid1 ovary tissues revealed the downregulation of FhRWP.The results indicated that FhARID1,as an upstream regulator of FhRWP,has an effect on the development of citrus axillary buds.Expression analysis of overexpressed leaves of FhARID1 lines showed that no significant up-regulation of FhRWP,indicating that FhARID1 is not the sole upstream regulatory factor of FhRWP.Only FhARID2 showed a correlation in expression with FhARID1 among the ARID-related genes,further supporting the notion that this gene may be involved in complex formation rather than acting alone.Yeast two-hybrid and MS/MS spectra further indicated that FhARID1 function requires casein kinase II-mediated post-transcriptional phosphorylation.This study elucidated the function of FhARID1 in citrus apomixis and axillary bud development,providing a fundamental basis for understanding the role of ARID-HMG-related genes.展开更多
Introduction:Diet intervention,especially supplementation with high-quality protein,is considered to be a critical strategy in sarcopenia.However,different sources and types of protein have different health impacts.Ob...Introduction:Diet intervention,especially supplementation with high-quality protein,is considered to be a critical strategy in sarcopenia.However,different sources and types of protein have different health impacts.Objectives:The aim of this study is to explore the differences in the ameliorative effects and mechanisms of different sources and types of proteins on sarcopenia,providing an optimal path for the prevention and treatment of sarcopenia.Methods:A sarcopenia model was established by intraperitoneal injection of dexamethasone(5 mg/kg).Sixty male C57BL/6 mice(8 months old)were randomly divided into the normal control,sarcopenia,goat whey protein,goat milk casein,bovine whey protein,and bovine milk casein groups.Animals were treated for 8 consecutive weeks.Organism-level and molecular phenotypes,16S rRNA gene sequencing,and untargeted metabolomics profiling based on GC-TOF/MS were employed to investigate the correlation between host metabolism,microbial metabolism,autophagy and inflammation and their influence on sarcopenia in C57BL/6 male mice.Results:All 4 proteins increased muscle mass,and goat whey protein improved muscle strength in sarcopenic mice.Goat and bovine milk proteins promoted muscle regeneration by increasing MyoD1 and MyoG expression,and the former had a more distinct effect in inducing autophagy and decreasing inflammation than the latter.In addition,goat whey protein and casein could modulate hostmicrobial arginine co-metabolism.Notably,goat milk proteins responded well to sarcopenia comorbidities,including sarcopenic obesity,osteosarcopenia,and osteoarthritis.Conclusion:The study confirmed that goat milk proteins were more effective than bovine milk proteins for the control of sarcopenia.Moreover,we found that whey protein and casein could modulate host-microbial arginine co-metabolism,which shows their potential as precision nutritional supplements for the management of sarcopenia.Our study provides theoretical support for the prevention and control of sarcopenia.展开更多
Insomnia is associated with neurotransmitters and intestinal dysbiosis.Though studies have demonstrated the ameliorative effects of milk hydrolysates on insomnia,the underlying mechanisms require further exploration.I...Insomnia is associated with neurotransmitters and intestinal dysbiosis.Though studies have demonstrated the ameliorative effects of milk hydrolysates on insomnia,the underlying mechanisms require further exploration.In this study,we investigated how papain hydrolysates of goat casein(CPH)and whey protein(WPH)affected mice's sleeplessness.Here,we show that CPH effectively improved the total sleep time in 12 h and restoring neurotransmitters(5-hydroxytryptamine(5-HT),γ-aminobutyric acid(GABA),dopamine(DA),and norepinephrine(NE))in mice.Further gut microbiota analysis revealed a significant increase in the relative abundance of Helicobacter and Escherichia-Shigella and a decrease in the relative abundance of Lactobacillus in the insomnia model mice(Model).Compared to the Model group,both CPH and WPH significantly increased the relative abundance of Akkermansia and Lactobacillus while lowering the relative abundance of Helicobacter and Escherichia-Shigella.Notably,while diazepam(DZP)increased mouse sleep duration,it also increased the relative abundance of Colidextribacter,Parasutterella,Muribaculaceae,and Prevotella.Additionally,the gene expression and protein expression of GABA_A receptor,cAMP-response element binding protein(CREB),and brain-derived neurotrophic factor(BDNF)were upregulated in the hypothalamus.We also discovered a link between intestinal gut microbiota and neurotransmitters.Overall,our results suggest that goat milk hydrolysates,especially CPH,can effectively improve insomnia,providing a theoretical basis for further experimentation and individualized designs.展开更多
Casein kinase 1(CK1)is an important member of the serine/threonine protein kinase family,playing a crucial role in various cellular processes,including cell cycle regulation,signal transduction,DNA repair,and circadia...Casein kinase 1(CK1)is an important member of the serine/threonine protein kinase family,playing a crucial role in various cellular processes,including cell cycle regulation,signal transduction,DNA repair,and circadian rhythm control.CK1 is also essential in the nervous system,where it regulates neuronal growth,differentiation,and synaptic plasticity.Studies have shown that CK1δ phosphorylates neuron-specific proteins to regulate axonal growth and synaptogenesis.Primary cilia are non-motile microtubule structures present on the surface of most mammalian cells.Recent studies have revealed their multiple roles in cellular physiology and development,and dysfunction of cilia can impact the development and function of the nervous system.CK1 has an important role in the formation and function of primary cilia.By regulating various signaling pathways and the phosphorylation status of proteins,CK1 affects the generation,maintenance,and signaling transduction of cilia.In this review,the relationship between CK1,primary cilia,and the nervous system was explored,focusing on how CK1 influences cilia to regulate the structure and function of the nervous system.展开更多
In this study,two series of foams based on tannic acid(TA),furfuryl alcohol(FA),soybean protein isolate(SPI),and casein(CA),namely TA–FA–SPI(TS series)and TA–FA–CA(TC series)were developed,and their properties wer...In this study,two series of foams based on tannic acid(TA),furfuryl alcohol(FA),soybean protein isolate(SPI),and casein(CA),namely TA–FA–SPI(TS series)and TA–FA–CA(TC series)were developed,and their properties were enhanced by adding poplar fibers(WF).From the samples produced,a complete set of characterization was performed including possible crosslinking reactions,morphology,mechanical properties,flame retardancy,thermal insulation and thermal stability.Fourier-transform infrared spectroscopy(FTIR)revealed possible covalent crosslinking among the components and hydrogen bonding between WF and the matrix.Viscosity results indicated that lower prepolymer viscosity led to lower apparent density,while WF addition reduced even more the density.Mechanical tests showed that the maximum compressive strengths were good,while WF improved the compressive strength by up to 56%.Scanning electron microscopy(SEM)showed uniform cell structures,but small open pores were observed.Two-dimensional(2D)CT scan images confirmed the good compatibility between WF and the matrix,with low anisotropy in the material.Friability tests indicated that WF decreased the pulverization ratio of the materials by up to 42%.Thermogravimetric analysis(TGA)showed good thermal stability of the materials up to 328°C.Vertical burning tests showed that the materials were self-extinguishing without residue(dripping).The lowest thermal conductivity was 0.04 W/m·K.These results suggest that these novel formaldehyde-free,high biomass content(95%–96%)foams and composite foams have high potential to replace traditional phenolic foams(PF)in applications such as construction,transportation,packaging,and thermal insulation.展开更多
Immunosuppressants currently approved for the treatment of autoimmune diseases and organ transplant rejection present diverse adverse effects that impair the life quality of patients.Therefore,the development of novel...Immunosuppressants currently approved for the treatment of autoimmune diseases and organ transplant rejection present diverse adverse effects that impair the life quality of patients.Therefore,the development of novel immunomodulators with high efficiency and low toxicity is essential.Ellagic acid(EA),a natural polyphenol compound widely distributed in berries,is metabolically transformed by gut microbiome to exert systemic health benefits.Here,we identified that intraperitoneal administration of EA with no cytotoxicity,beyond its wellknown oral metabolic fate,effectively decreased clinical severity and central nervous system(CNS)inflammation/demyelination in experimental autoimmune encephalomyelitis,a mouse model of an autoimmune disease multiple sclerosis.Interestingly,intraperitoneal EA administration at incredibly low doses(0.1 mg/(kg·day))is dose-sparing with fingolimod(FTY720),the first FDA-approved oral drug for MS.In addition,intraperitoneal EA also ameliorated the brain damage in a neuromyelitis optica(NMO)model,and significantly prevented the immune rejection of allograft skin graft.Evidence from pharmacological studies combined with RNA-seq indicate that prototype EA functions by a mechanism that involves direct inhibition of casein kinase II(CKII)to suppress the expression of IL-17 and promote the expression of Cpt1a to regulate T helper cell 17 differentiation.In conclusion,our study demonstrates that the prototype EA entering the blood circulation acts as a novel therapeutic immunomodulator for the treatment of autoimmune diseases and transplant rejection through the CKII-mediated Janus kinase/signal transducer and activator of transcription 3 Cpt1a signaling pathway.展开更多
Intraocular pressure elevation can induce retinal ganglion cell death and is a clinically reversible risk factor for glaucoma,the leading cause of irreversible blindness.We previously demonstrated that casein kinase-2...Intraocular pressure elevation can induce retinal ganglion cell death and is a clinically reversible risk factor for glaucoma,the leading cause of irreversible blindness.We previously demonstrated that casein kinase-2 inhibition can promote retinal ganglion cell survival and axonal regeneration in rats after optic nerve injury.To investigate the underlying mechanism,in the current study we increased the intraocular pressure of adult rats to 75 mmHg for 2 hours and then administered a casein kinase-2 inhibitor(4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole)by intravitreal injection.We found that intravitreal injection of 4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole promoted retinal ganglion cell survival and reduced the number of infiltrating macrophages.Transcriptomic analysis showed that the mitogen activated protein kinase signaling pathway was involved in the response to intraocular pressure elevation but was not modulated by the casein kinase-2 inhibitors.Furthermore,casein kinase-2 inhibition downregulated the expression of genes(Cck,Htrsa,Nef1,Htrlb,Prph,Chat,Slc18a3,Slc5a7,Scn1b,Crybb2,Tsga10ip,and Vstm21)involved in intraocular pressure elevation.Our data indicate that inhibition of casein kinase-2 can enhance retinal ganglion cell survival in rats after acute intraocular pressure elevation via macrophage inactivation.展开更多
Tiller number and grain size are important agronomic traits that determine grain yield in rice.Here,we demonstrate that DEFECTIVE TILLER GROWTH 1(DTG1),a member of the casein kinase 1 protein family,exerts a co-regula...Tiller number and grain size are important agronomic traits that determine grain yield in rice.Here,we demonstrate that DEFECTIVE TILLER GROWTH 1(DTG1),a member of the casein kinase 1 protein family,exerts a co-regulatory effect on tiller number and grain size.We identified a single amino acid substitution in DTG1(I357K)that caused a decrease in tiller number and an increase in grain size in NIL-dtg1.Genetic analyses revealed that DTG1 plays a pivotal role in regulation of tillering and grain size.The DTG1^(I357K) allelic variant exhibited robust functionality in suppressing tillering.We show that DTG1 is preferentially expressed in tiller buds and young panicles,and negatively regulates grain size by restricting cell proliferation in spikelet hulls.We further confirm that DTG1 functioned in grain size regulation by directly interacting with Grain Width 2(GW2),a critical grain size regulator in rice.The CRISPR/Cas9-mediated elimination of DTG1 significantly enhanced tiller number and grain size,thereby increasing rice grain yield under field conditions,thus highlighting potential value of DTG1 in rice breeding.展开更多
Introduction:The regulation of thyroid-stimulating hormone(TSH)synthesis involves neurotransmitters,with melatonin being a subject of ongoing debate.TSH transcription,synthesis,and secretion from the pituitary pars di...Introduction:The regulation of thyroid-stimulating hormone(TSH)synthesis involves neurotransmitters,with melatonin being a subject of ongoing debate.TSH transcription,synthesis,and secretion from the pituitary pars distalis(PD)is primarily regulated in a photoperiodic manner by thyrotropin-releasing hormone(TRH).In contrast,in the pituitary pars tuberalis(PT),mRNA transcription and alpha/beta chain synthesis,but not secretion,of a TSHlike product is regulated by melatonin.Conversely,non-photoperiodic melatonin might also affect the secretion of a TSH-like product from the PT.Nevertheless,the impact of exogenous melatonin on the underlying PD-TSH synthesis remains unclear.Casein kinase 1α(CK1α)plays a negative regulatory role in TSH synthesis in the mouse pituitary.Objective:We investigated whether non-photoperiodic melatonin affects PD-TSH synthesis through its interaction with CK1α.Methods:Immunohistochemistry and immunofluorescence staining detected the colocalization of the melatonin receptor MT1 with CK1αand TSH-βin the PD.RT-qPCR,western blotting,and ELISA revealed the effect of melatonin on Tshb mRNA,MTNR1A mRNA,Csnk1a1 mRNA,CK1αprotein,MT1 protein,and TSH levels.Results:Robust colocalization of the melatonin receptor MT1 with CK1αand TSH-βin the PD.Tshb mRNA and CK1αprotein expression levels peaked at opposite phases of the 24-h light:dark cycle.Exogenous melatonin administration promoted pituitary TSH synthesis,while concurrently inhibiting CK1αactivity.The upregulation of endogenous CK1αactivity in primary pituitary cells significantly blunted the melatonin stimulatory impact on Tshb mRNA and TSH levels.Mechanistically,the CK1αagonist pyrvinium abrogated melatonin-induced activation of p-PKC and p-CREB expression in vitro.Conclusion:The CK1α/PKC signaling pathway mediates the regulation of melatonin in pituitary TSH synthesis.We demonstrate an important theoretical and experimental basis for understanding the mechanism of endocrine system diseases caused by abnormal TSH synthesis in the pituitary.展开更多
Objective Machado-Joseph disease （MJD）/Spinocerebellar ataxia type 3 （SCA3） is an autosomal dominant neurodegenerative disorder caused by an expansion of polyglutamine tract near the C-terminus of the MJD1 gene pr...Objective Machado-Joseph disease （MJD）/Spinocerebellar ataxia type 3 （SCA3） is an autosomal dominant neurodegenerative disorder caused by an expansion of polyglutamine tract near the C-terminus of the MJD1 gene product, ataxin-3. The precise mechanism of the MJD/SCA3 pathogenesis remains unclear. A growing body of evidence demonstrates that phosphorylation plays an important role in the pathogenesis of many neurodegenerative diseases. However, few kinases are known to phosphorylate ataxin-3. The present study is to explore whether ataxin-3 is a substrate of casein kinase 2 （CK2）. Methods The interaction between ataxin-3 and CK2 was identified by glutathione S-transferase （GST） pull-down assay and co-immunoprecipition assay. The phosphorylation of ataxin-3 by CK2 was measured by in vitro phosphorylation assays. Results （1） Both wild type and expanded ataxin-3 interacted with CK2α and CK2β in vitro. （2） In 293 cells, both wild type and expanded ataxin-3 interacted with CK2β, but not CK2α. （3） CK2 phosphorylated wild type and expanded ataxin-3. Conclusion Ataxin-3 is a substrate of protein kinase CK2.展开更多
The kinetics of casein tryptic hydrolysis to prepare activepeptides was investigated. Taking into account the reaction mechanismincluding single substrate hydrolysis, irreversible enzymeinactivation, and substrate inh...The kinetics of casein tryptic hydrolysis to prepare activepeptides was investigated. Taking into account the reaction mechanismincluding single substrate hydrolysis, irreversible enzymeinactivation, and substrate inhibition, a set of exponentialequations was established to characterize the enzymatic hydrolysiscurves. The verification was carried out by a series of experimentalresults and indicated that the average regressive error was less than5/100. According to the proposed kinetic model, the kinetic constantsand thermodynamic constants of the reaction system were alsocalculated.展开更多
Background:High-protein diets can increase the colonic health risks.A moderate reduction of dietary crude-protein(CP)level can improve the colonic bacterial community and mucosal immunity of pigs.However,greatly reduc...Background:High-protein diets can increase the colonic health risks.A moderate reduction of dietary crude-protein(CP)level can improve the colonic bacterial community and mucosal immunity of pigs.However,greatly reducing the dietary CP level,even supplemented with all amino acids(AAs),detrimentally affects the colonic health,which may be due to the lack of protein-derived peptides.Therefore,this study evaluated the effects of supplementation of casein hydrolysate(peptide source)in low-protein(LP)diets,in comparison with AAs supplementation,on the colonic microbiota,microbial metabolites and mucosal immunity in pigs,aiming to determine whether a supplementation of casein hydrolysate can improve colonic health under very LP level.Twenty-one pigs(initial BW 19.90±1.00 kg,63±1 days of age)were assigned to three groups and fed with control diet(16%CP),LP diets(13%CP)supplemented with free AAs(LPA)or casein hydrolysate(LPC)for 4 weeks.Results:Compared with control diet,LPA and LPC diet decreased the relative abundance of Streptococcus and Escherichia coli,and LPC diet further decreased the relative abundance of Proteobacteria.LPC diet also increased the relative abundance of Lactobacillus reuteri.Both LP diets decreased concentrations of ammonia and cadaverine,and LPC diet also reduced concentrations of putrescine,phenol and indole.Moreover,LPC diet increased total short-chain fatty acid concentration.In comparison with control diet,both LP diets decreased protein expressions of Toll-like receptor-4,nuclear factor-κB,interleukin-1βand tumor necrosis factor-α,and LPC diet further decreased protein expressions of nucleotide-binding oligomerization domain protein-1 and interferon-γ.LPC diet also increased protein expressions of G-protein coupled receptor-43,interleukin-4,transforming growth factor-β,immunoglobulin A and mucin-4,which are indicators for mucosal defense activity.Conclusions:The results showed that supplementing casein hydrolysate showed beneficial effects on the colonic microbiota and mucosal immunity and barrier function in comparison with supplementing free AAs in LP diets.These findings may provide new framework for future nutritional interventions for colon health in pigs.展开更多
AIM: To characterize the nuclear import of hepatitis B virus (HBV) polymerase (P) and its relevance for the viral life cycle.METHODS: Sequence analysis was performed to predict functional motives within P. Phosphoryla...AIM: To characterize the nuclear import of hepatitis B virus (HBV) polymerase (P) and its relevance for the viral life cycle.METHODS: Sequence analysis was performed to predict functional motives within P. Phosphorylation of P was analyzed by in vitro phosphorylation. Phosphorylation site and nuclear localization signal (NLS) were destroyed by site directed mutagenesis. Functionality of the identified NLS was analyzed by confocal fluorescence microscopy and characterizing the karyopherin binding. Relevance of the structural motives for viral life cycle was studied by infection of primary Tupaia hepatocytes with HBV.RESULTS: We identified by sequence alignment and functional experiments a conserved bipartite NLS containing a casein kinase II (CKII) phosphorylation site located within the terminal protein domain (TP) of the HBV polymerase. Inhibition of CKII impairs the functionality of this NLS and thereby prevents the nuclear import of the polymerase. Binding of the import factor karyopherin-α2 to the polymerase depends on its CKII-mediated phosphorylation of the bipartite NLS. In HBV-infected primary Tupaia hepatocytes CKII inhibition in the early phase (post entry phase) of the infection process prevents the establishment of the infection.CONCLUSION: Based on these data it is suggested that during HBV infection the final import of the genome complex into the nucleus is mediated by a novel bipartite NLS localized in the TP domain of HBV polymerase.展开更多
There is no study on food-derived peptide with both anticoagulant and angiotensin I-converting enzyme inhibitory (ACEI) activities yet. In this work, the anticoagulant and ACEI activities of the casein hydrolysates re...There is no study on food-derived peptide with both anticoagulant and angiotensin I-converting enzyme inhibitory (ACEI) activities yet. In this work, the anticoagulant and ACEI activities of the casein hydrolysates released by pepsin digestion were evaluated for the first time to the best of our knowledge. Results indicated that the casein hydrolysate exhibited potent anticoagulant activity by prolonging the thrombin time (TT) and the activated partial thromboplastin time (APTT). Compared with control samples, at 10 mg/mL, the TT and APTT of casein hydrolysate were 186.0 % ± 6.6 % and 163.5 % ± 7.4 %, respectively. The casein hydrolysate also showed a strong ACEI activity with an IC50 value of 1.775 mg/mL. The components of the bioactive casein hydrolysate were analyzed by nanoscale liquid chromatography quadrupole time-of-flight tandem mass spectrometry (NanoLC-Q-TOF-MS/MS). Total of 115 peptides were identified, among which 34, 9, 55 and 17 peptides were derived from α_(s1-), α_(s2-), β-, and κ-casein, respectively. The results of PeptideRanker and PepSite 2 analysis showed that 6 peptides (FRQFYQL, NENLLRF, NPWDQVKR, PVVVPPFLQ, PVRGPFPIIV, and ARHPHPHLSF) have both ACEI and anticoagulant activities by binding to the active sites of ACE and thrombin. This study indicated that casein is a potential functional food supplement that can be used for medical purposes.展开更多
Here, we describe the use of monolayers of intestinal epithelial cells derived from intestinal organoids and transcriptomics to investigate the direct effects of dietary protein sources on epithelial function. Mechani...Here, we describe the use of monolayers of intestinal epithelial cells derived from intestinal organoids and transcriptomics to investigate the direct effects of dietary protein sources on epithelial function. Mechanically dissociated 3 D organoids of mouse duodenum were used to generate a polarized epithelium containing all cell types found in the tissue of origin. The organoid-derived cell monolayers were exposed to 4%(w/v) of ‘undigested(non-hydrolysed)-soluble' fraction of protein sources used as feed ingredients [soybean meal(SBM) and casein], or alternative protein sources(spray dried plasma protein, and yellow meal worm), or controls for 6 h prior to RNA isolation and transcriptomics. All protein sources altered expression of unique biological processes in the epithelial cells. Exposure of intestinal organoids to SBM downregulated expression of retinol and retinoid metabolic processes as well as cholesterol and lipid biosynthetic pathways, consistent with the reported hypotriglyceridaemic effect of soy protein in vivo. These findings support the use of intestinal organoids as models to evaluate complex interactions between dietary ingredients and the intestinal epithelium and highlights some unique host effects of alternative protein sources in animal feed and potentially human food.展开更多
基金International Science&Technology Cooperation Program of China(2011DFA32550)Ministry of Science and Technology of China(2012BAD12B08)
文摘The effects of heat treatment(heating temperature and pH) on the structures and emulsifying properties of caseins were systematically studied by spectroscopy.Heat treatment from 60to 100℃resulted in an increase in their fluorescence intensity,hydrodynamic diameter,turbidity and emulsifying activity index,but decreased the size polydispersity of caseins.In the pH range of 5.5to 7.0,the fluorescence intensity,hydrodynamic diameter,turbidity and emulsifying properties decreased with increased heating pH,but the size polydispersity of caseins increased with increased pH.The relationship between the surface fluorescence intensity and emulsifying activity was also investigated,revealing a correlation coefficient of 0.90.These results suggested that heat treatment could be used to modify the structures and emulsifying properties of caseins by appropriately selecting heating conditions.
文摘Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.
文摘This paper carried out a comparative analysis of different types of electrophoretic systems which were used for the analysis of casein complex from cow milk (polyacrylamide gel electrophoresis: for the neutral and acidic native conditions, in gradient variant, the presence of sodium dodecyl sulphate or with including urea). Taking into attention the separation efficiency, complexity of electrophoretic system, the impact of system components, we have selected the anode system of the homogeneous gel in the presence of urea as the basis for the preparation of casein fractions. It also changed the composition and the structure of the electrophoretic apparatus. The changes allow purification of casein fractions up to several grams during one stage of treatment (for 5 hours). The purified casein fractions were tested for the homogeneity and have been recommended for using in the biomedical researches, including the processes of the formation of the bioactive peptides.
基金funded by the National Key Research and Development Program of China(Grant No.2022YFF1003100)Modern Citrus Industry Technology System of China(Grant No.CARS-26).
文摘Polyembryony has posed a significant impediment to the advancement of citrus hybrid breeding.FhRWP is widely regarded as a pivotal factor governing asexual reproduction in citrus,and prior research has demonstrated that FhARID1,acting as an upstream regulator,modulates FhRWP expression.In this study,we performed a genome-wide characterization of the ARID-HMG-related genes using the short juvenile minicitrus Fortunella hindsii.A total of 20 ARID-HMG-related genes were identified.Protein interaction network and enrichment analysis suggested that ARID-HMG-related proteins might might be involved in chromatin remodeling complexes.Knockout of FhARID1 in F.hindsii did not induce the conversion from polyembryony to monoembryony.However,fharid1 plants in T1 generation exhibited abnormal proliferation at axillary buds,which is similar to phenotype of fhrwp plants.Expression analysis of fharid1 ovary tissues revealed the downregulation of FhRWP.The results indicated that FhARID1,as an upstream regulator of FhRWP,has an effect on the development of citrus axillary buds.Expression analysis of overexpressed leaves of FhARID1 lines showed that no significant up-regulation of FhRWP,indicating that FhARID1 is not the sole upstream regulatory factor of FhRWP.Only FhARID2 showed a correlation in expression with FhARID1 among the ARID-related genes,further supporting the notion that this gene may be involved in complex formation rather than acting alone.Yeast two-hybrid and MS/MS spectra further indicated that FhARID1 function requires casein kinase II-mediated post-transcriptional phosphorylation.This study elucidated the function of FhARID1 in citrus apomixis and axillary bud development,providing a fundamental basis for understanding the role of ARID-HMG-related genes.
基金supported by the Beijing Natural Science Foundation(7232236)the National Key R&D Program of China(2022YFF1100104)。
文摘Introduction:Diet intervention,especially supplementation with high-quality protein,is considered to be a critical strategy in sarcopenia.However,different sources and types of protein have different health impacts.Objectives:The aim of this study is to explore the differences in the ameliorative effects and mechanisms of different sources and types of proteins on sarcopenia,providing an optimal path for the prevention and treatment of sarcopenia.Methods:A sarcopenia model was established by intraperitoneal injection of dexamethasone(5 mg/kg).Sixty male C57BL/6 mice(8 months old)were randomly divided into the normal control,sarcopenia,goat whey protein,goat milk casein,bovine whey protein,and bovine milk casein groups.Animals were treated for 8 consecutive weeks.Organism-level and molecular phenotypes,16S rRNA gene sequencing,and untargeted metabolomics profiling based on GC-TOF/MS were employed to investigate the correlation between host metabolism,microbial metabolism,autophagy and inflammation and their influence on sarcopenia in C57BL/6 male mice.Results:All 4 proteins increased muscle mass,and goat whey protein improved muscle strength in sarcopenic mice.Goat and bovine milk proteins promoted muscle regeneration by increasing MyoD1 and MyoG expression,and the former had a more distinct effect in inducing autophagy and decreasing inflammation than the latter.In addition,goat whey protein and casein could modulate hostmicrobial arginine co-metabolism.Notably,goat milk proteins responded well to sarcopenia comorbidities,including sarcopenic obesity,osteosarcopenia,and osteoarthritis.Conclusion:The study confirmed that goat milk proteins were more effective than bovine milk proteins for the control of sarcopenia.Moreover,we found that whey protein and casein could modulate host-microbial arginine co-metabolism,which shows their potential as precision nutritional supplements for the management of sarcopenia.Our study provides theoretical support for the prevention and control of sarcopenia.
基金supported by the National First-class Discipline Program of Food Science and Technology(JUFSTR20180204)。
文摘Insomnia is associated with neurotransmitters and intestinal dysbiosis.Though studies have demonstrated the ameliorative effects of milk hydrolysates on insomnia,the underlying mechanisms require further exploration.In this study,we investigated how papain hydrolysates of goat casein(CPH)and whey protein(WPH)affected mice's sleeplessness.Here,we show that CPH effectively improved the total sleep time in 12 h and restoring neurotransmitters(5-hydroxytryptamine(5-HT),γ-aminobutyric acid(GABA),dopamine(DA),and norepinephrine(NE))in mice.Further gut microbiota analysis revealed a significant increase in the relative abundance of Helicobacter and Escherichia-Shigella and a decrease in the relative abundance of Lactobacillus in the insomnia model mice(Model).Compared to the Model group,both CPH and WPH significantly increased the relative abundance of Akkermansia and Lactobacillus while lowering the relative abundance of Helicobacter and Escherichia-Shigella.Notably,while diazepam(DZP)increased mouse sleep duration,it also increased the relative abundance of Colidextribacter,Parasutterella,Muribaculaceae,and Prevotella.Additionally,the gene expression and protein expression of GABA_A receptor,cAMP-response element binding protein(CREB),and brain-derived neurotrophic factor(BDNF)were upregulated in the hypothalamus.We also discovered a link between intestinal gut microbiota and neurotransmitters.Overall,our results suggest that goat milk hydrolysates,especially CPH,can effectively improve insomnia,providing a theoretical basis for further experimentation and individualized designs.
文摘Casein kinase 1(CK1)is an important member of the serine/threonine protein kinase family,playing a crucial role in various cellular processes,including cell cycle regulation,signal transduction,DNA repair,and circadian rhythm control.CK1 is also essential in the nervous system,where it regulates neuronal growth,differentiation,and synaptic plasticity.Studies have shown that CK1δ phosphorylates neuron-specific proteins to regulate axonal growth and synaptogenesis.Primary cilia are non-motile microtubule structures present on the surface of most mammalian cells.Recent studies have revealed their multiple roles in cellular physiology and development,and dysfunction of cilia can impact the development and function of the nervous system.CK1 has an important role in the formation and function of primary cilia.By regulating various signaling pathways and the phosphorylation status of proteins,CK1 affects the generation,maintenance,and signaling transduction of cilia.In this review,the relationship between CK1,primary cilia,and the nervous system was explored,focusing on how CK1 influences cilia to regulate the structure and function of the nervous system.
基金funded by the Natural Sciences and Engineering Research Council of Canada(NSERC)for financial support,Grant Nos.CG125664 and GF136078.
文摘In this study,two series of foams based on tannic acid(TA),furfuryl alcohol(FA),soybean protein isolate(SPI),and casein(CA),namely TA–FA–SPI(TS series)and TA–FA–CA(TC series)were developed,and their properties were enhanced by adding poplar fibers(WF).From the samples produced,a complete set of characterization was performed including possible crosslinking reactions,morphology,mechanical properties,flame retardancy,thermal insulation and thermal stability.Fourier-transform infrared spectroscopy(FTIR)revealed possible covalent crosslinking among the components and hydrogen bonding between WF and the matrix.Viscosity results indicated that lower prepolymer viscosity led to lower apparent density,while WF addition reduced even more the density.Mechanical tests showed that the maximum compressive strengths were good,while WF improved the compressive strength by up to 56%.Scanning electron microscopy(SEM)showed uniform cell structures,but small open pores were observed.Two-dimensional(2D)CT scan images confirmed the good compatibility between WF and the matrix,with low anisotropy in the material.Friability tests indicated that WF decreased the pulverization ratio of the materials by up to 42%.Thermogravimetric analysis(TGA)showed good thermal stability of the materials up to 328°C.Vertical burning tests showed that the materials were self-extinguishing without residue(dripping).The lowest thermal conductivity was 0.04 W/m·K.These results suggest that these novel formaldehyde-free,high biomass content(95%–96%)foams and composite foams have high potential to replace traditional phenolic foams(PF)in applications such as construction,transportation,packaging,and thermal insulation.
基金supported by the Chinese National Natural Science Foundation(92268118,82071396,82271199)the Shaanxi Provincial Key R&D Foundation(2021ZDLSF03-09)+3 种基金the Scientific Research Program Funded by Shaanxi Provincial Education Department(22JK0550)the Fundamental Research Funds for the Central Universities(GK202201013,GK202202006,GK202305001)the Excellent Graduate Training Program of Shaanxi Normal University(LHRCYB23003)Tian Jiabing Scholar Program.
文摘Immunosuppressants currently approved for the treatment of autoimmune diseases and organ transplant rejection present diverse adverse effects that impair the life quality of patients.Therefore,the development of novel immunomodulators with high efficiency and low toxicity is essential.Ellagic acid(EA),a natural polyphenol compound widely distributed in berries,is metabolically transformed by gut microbiome to exert systemic health benefits.Here,we identified that intraperitoneal administration of EA with no cytotoxicity,beyond its wellknown oral metabolic fate,effectively decreased clinical severity and central nervous system(CNS)inflammation/demyelination in experimental autoimmune encephalomyelitis,a mouse model of an autoimmune disease multiple sclerosis.Interestingly,intraperitoneal EA administration at incredibly low doses(0.1 mg/(kg·day))is dose-sparing with fingolimod(FTY720),the first FDA-approved oral drug for MS.In addition,intraperitoneal EA also ameliorated the brain damage in a neuromyelitis optica(NMO)model,and significantly prevented the immune rejection of allograft skin graft.Evidence from pharmacological studies combined with RNA-seq indicate that prototype EA functions by a mechanism that involves direct inhibition of casein kinase II(CKII)to suppress the expression of IL-17 and promote the expression of Cpt1a to regulate T helper cell 17 differentiation.In conclusion,our study demonstrates that the prototype EA entering the blood circulation acts as a novel therapeutic immunomodulator for the treatment of autoimmune diseases and transplant rejection through the CKII-mediated Janus kinase/signal transducer and activator of transcription 3 Cpt1a signaling pathway.
基金supported by the National Natural Science Foundation of China,Nos.81570849,81100931the Natural Science Foundation of Guangdong Province of China,Nos.2015A030313446,2020A1515011413(all to LPC).
文摘Intraocular pressure elevation can induce retinal ganglion cell death and is a clinically reversible risk factor for glaucoma,the leading cause of irreversible blindness.We previously demonstrated that casein kinase-2 inhibition can promote retinal ganglion cell survival and axonal regeneration in rats after optic nerve injury.To investigate the underlying mechanism,in the current study we increased the intraocular pressure of adult rats to 75 mmHg for 2 hours and then administered a casein kinase-2 inhibitor(4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole)by intravitreal injection.We found that intravitreal injection of 4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole promoted retinal ganglion cell survival and reduced the number of infiltrating macrophages.Transcriptomic analysis showed that the mitogen activated protein kinase signaling pathway was involved in the response to intraocular pressure elevation but was not modulated by the casein kinase-2 inhibitors.Furthermore,casein kinase-2 inhibition downregulated the expression of genes(Cck,Htrsa,Nef1,Htrlb,Prph,Chat,Slc18a3,Slc5a7,Scn1b,Crybb2,Tsga10ip,and Vstm21)involved in intraocular pressure elevation.Our data indicate that inhibition of casein kinase-2 can enhance retinal ganglion cell survival in rats after acute intraocular pressure elevation via macrophage inactivation.
基金supported by the Sichuan Science and Technology Support Project (2022ZDZX0012,2021YFYZ0016,2023YFN0007,2021YFYZ0027)the National Natural Science Foundation of China (32171966,U23A20180).
文摘Tiller number and grain size are important agronomic traits that determine grain yield in rice.Here,we demonstrate that DEFECTIVE TILLER GROWTH 1(DTG1),a member of the casein kinase 1 protein family,exerts a co-regulatory effect on tiller number and grain size.We identified a single amino acid substitution in DTG1(I357K)that caused a decrease in tiller number and an increase in grain size in NIL-dtg1.Genetic analyses revealed that DTG1 plays a pivotal role in regulation of tillering and grain size.The DTG1^(I357K) allelic variant exhibited robust functionality in suppressing tillering.We show that DTG1 is preferentially expressed in tiller buds and young panicles,and negatively regulates grain size by restricting cell proliferation in spikelet hulls.We further confirm that DTG1 functioned in grain size regulation by directly interacting with Grain Width 2(GW2),a critical grain size regulator in rice.The CRISPR/Cas9-mediated elimination of DTG1 significantly enhanced tiller number and grain size,thereby increasing rice grain yield under field conditions,thus highlighting potential value of DTG1 in rice breeding.
基金supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Introduction:The regulation of thyroid-stimulating hormone(TSH)synthesis involves neurotransmitters,with melatonin being a subject of ongoing debate.TSH transcription,synthesis,and secretion from the pituitary pars distalis(PD)is primarily regulated in a photoperiodic manner by thyrotropin-releasing hormone(TRH).In contrast,in the pituitary pars tuberalis(PT),mRNA transcription and alpha/beta chain synthesis,but not secretion,of a TSHlike product is regulated by melatonin.Conversely,non-photoperiodic melatonin might also affect the secretion of a TSH-like product from the PT.Nevertheless,the impact of exogenous melatonin on the underlying PD-TSH synthesis remains unclear.Casein kinase 1α(CK1α)plays a negative regulatory role in TSH synthesis in the mouse pituitary.Objective:We investigated whether non-photoperiodic melatonin affects PD-TSH synthesis through its interaction with CK1α.Methods:Immunohistochemistry and immunofluorescence staining detected the colocalization of the melatonin receptor MT1 with CK1αand TSH-βin the PD.RT-qPCR,western blotting,and ELISA revealed the effect of melatonin on Tshb mRNA,MTNR1A mRNA,Csnk1a1 mRNA,CK1αprotein,MT1 protein,and TSH levels.Results:Robust colocalization of the melatonin receptor MT1 with CK1αand TSH-βin the PD.Tshb mRNA and CK1αprotein expression levels peaked at opposite phases of the 24-h light:dark cycle.Exogenous melatonin administration promoted pituitary TSH synthesis,while concurrently inhibiting CK1αactivity.The upregulation of endogenous CK1αactivity in primary pituitary cells significantly blunted the melatonin stimulatory impact on Tshb mRNA and TSH levels.Mechanistically,the CK1αagonist pyrvinium abrogated melatonin-induced activation of p-PKC and p-CREB expression in vitro.Conclusion:The CK1α/PKC signaling pathway mediates the regulation of melatonin in pituitary TSH synthesis.We demonstrate an important theoretical and experimental basis for understanding the mechanism of endocrine system diseases caused by abnormal TSH synthesis in the pituitary.
基金the National Natural Sciences Foundation of China (No. 30770664)a grant from Educational Committee of Anhui Province, China (No. ZD2008008-2).
文摘Objective Machado-Joseph disease （MJD）/Spinocerebellar ataxia type 3 （SCA3） is an autosomal dominant neurodegenerative disorder caused by an expansion of polyglutamine tract near the C-terminus of the MJD1 gene product, ataxin-3. The precise mechanism of the MJD/SCA3 pathogenesis remains unclear. A growing body of evidence demonstrates that phosphorylation plays an important role in the pathogenesis of many neurodegenerative diseases. However, few kinases are known to phosphorylate ataxin-3. The present study is to explore whether ataxin-3 is a substrate of casein kinase 2 （CK2）. Methods The interaction between ataxin-3 and CK2 was identified by glutathione S-transferase （GST） pull-down assay and co-immunoprecipition assay. The phosphorylation of ataxin-3 by CK2 was measured by in vitro phosphorylation assays. Results （1） Both wild type and expanded ataxin-3 interacted with CK2α and CK2β in vitro. （2） In 293 cells, both wild type and expanded ataxin-3 interacted with CK2β, but not CK2α. （3） CK2 phosphorylated wild type and expanded ataxin-3. Conclusion Ataxin-3 is a substrate of protein kinase CK2.
基金Supported by National Natural Science Foundation of China (No. 20276052) and Tianjin Science & Technology Commission (No. 023105411).
文摘The kinetics of casein tryptic hydrolysis to prepare activepeptides was investigated. Taking into account the reaction mechanismincluding single substrate hydrolysis, irreversible enzymeinactivation, and substrate inhibition, a set of exponentialequations was established to characterize the enzymatic hydrolysiscurves. The verification was carried out by a series of experimentalresults and indicated that the average regressive error was less than5/100. According to the proposed kinetic model, the kinetic constantsand thermodynamic constants of the reaction system were alsocalculated.
基金supported by National Key Basic Research Program of China(2013CB127300)Natural Science Foundation of China(31430082).
文摘Background:High-protein diets can increase the colonic health risks.A moderate reduction of dietary crude-protein(CP)level can improve the colonic bacterial community and mucosal immunity of pigs.However,greatly reducing the dietary CP level,even supplemented with all amino acids(AAs),detrimentally affects the colonic health,which may be due to the lack of protein-derived peptides.Therefore,this study evaluated the effects of supplementation of casein hydrolysate(peptide source)in low-protein(LP)diets,in comparison with AAs supplementation,on the colonic microbiota,microbial metabolites and mucosal immunity in pigs,aiming to determine whether a supplementation of casein hydrolysate can improve colonic health under very LP level.Twenty-one pigs(initial BW 19.90±1.00 kg,63±1 days of age)were assigned to three groups and fed with control diet(16%CP),LP diets(13%CP)supplemented with free AAs(LPA)or casein hydrolysate(LPC)for 4 weeks.Results:Compared with control diet,LPA and LPC diet decreased the relative abundance of Streptococcus and Escherichia coli,and LPC diet further decreased the relative abundance of Proteobacteria.LPC diet also increased the relative abundance of Lactobacillus reuteri.Both LP diets decreased concentrations of ammonia and cadaverine,and LPC diet also reduced concentrations of putrescine,phenol and indole.Moreover,LPC diet increased total short-chain fatty acid concentration.In comparison with control diet,both LP diets decreased protein expressions of Toll-like receptor-4,nuclear factor-κB,interleukin-1βand tumor necrosis factor-α,and LPC diet further decreased protein expressions of nucleotide-binding oligomerization domain protein-1 and interferon-γ.LPC diet also increased protein expressions of G-protein coupled receptor-43,interleukin-4,transforming growth factor-β,immunoglobulin A and mucin-4,which are indicators for mucosal defense activity.Conclusions:The results showed that supplementing casein hydrolysate showed beneficial effects on the colonic microbiota and mucosal immunity and barrier function in comparison with supplementing free AAs in LP diets.These findings may provide new framework for future nutritional interventions for colon health in pigs.
文摘AIM: To characterize the nuclear import of hepatitis B virus (HBV) polymerase (P) and its relevance for the viral life cycle.METHODS: Sequence analysis was performed to predict functional motives within P. Phosphorylation of P was analyzed by in vitro phosphorylation. Phosphorylation site and nuclear localization signal (NLS) were destroyed by site directed mutagenesis. Functionality of the identified NLS was analyzed by confocal fluorescence microscopy and characterizing the karyopherin binding. Relevance of the structural motives for viral life cycle was studied by infection of primary Tupaia hepatocytes with HBV.RESULTS: We identified by sequence alignment and functional experiments a conserved bipartite NLS containing a casein kinase II (CKII) phosphorylation site located within the terminal protein domain (TP) of the HBV polymerase. Inhibition of CKII impairs the functionality of this NLS and thereby prevents the nuclear import of the polymerase. Binding of the import factor karyopherin-α2 to the polymerase depends on its CKII-mediated phosphorylation of the bipartite NLS. In HBV-infected primary Tupaia hepatocytes CKII inhibition in the early phase (post entry phase) of the infection process prevents the establishment of the infection.CONCLUSION: Based on these data it is suggested that during HBV infection the final import of the genome complex into the nucleus is mediated by a novel bipartite NLS localized in the TP domain of HBV polymerase.
基金The China Postdoctoral Science Foundation(2019M661072)the Basic Research Program of Liaoning Education Department(2017J080)the National Natural Science Foundation of China(31771926)funded this study.
文摘There is no study on food-derived peptide with both anticoagulant and angiotensin I-converting enzyme inhibitory (ACEI) activities yet. In this work, the anticoagulant and ACEI activities of the casein hydrolysates released by pepsin digestion were evaluated for the first time to the best of our knowledge. Results indicated that the casein hydrolysate exhibited potent anticoagulant activity by prolonging the thrombin time (TT) and the activated partial thromboplastin time (APTT). Compared with control samples, at 10 mg/mL, the TT and APTT of casein hydrolysate were 186.0 % ± 6.6 % and 163.5 % ± 7.4 %, respectively. The casein hydrolysate also showed a strong ACEI activity with an IC50 value of 1.775 mg/mL. The components of the bioactive casein hydrolysate were analyzed by nanoscale liquid chromatography quadrupole time-of-flight tandem mass spectrometry (NanoLC-Q-TOF-MS/MS). Total of 115 peptides were identified, among which 34, 9, 55 and 17 peptides were derived from α_(s1-), α_(s2-), β-, and κ-casein, respectively. The results of PeptideRanker and PepSite 2 analysis showed that 6 peptides (FRQFYQL, NENLLRF, NPWDQVKR, PVVVPPFLQ, PVRGPFPIIV, and ARHPHPHLSF) have both ACEI and anticoagulant activities by binding to the active sites of ACE and thrombin. This study indicated that casein is a potential functional food supplement that can be used for medical purposes.
基金the financial support from the Wageningen University&Research “IPOP Customized Nutrition” program(grant no.4417801270)financed by Wageningen University&Research,the Dutch Ministry of Economic Affairs, Agriculture&Innovation, the graduate school Wageningen Institute of Animal Science(WIAS)+1 种基金industrial partners Trouw Nutrition, The Netherlands and Darling Ingredient International, The Netherlandssupported by the applied and engineering sciences division of The Netherlands Organisation for Scientific Research(NWO project 14935)and DSM Nutritional Products。
文摘Here, we describe the use of monolayers of intestinal epithelial cells derived from intestinal organoids and transcriptomics to investigate the direct effects of dietary protein sources on epithelial function. Mechanically dissociated 3 D organoids of mouse duodenum were used to generate a polarized epithelium containing all cell types found in the tissue of origin. The organoid-derived cell monolayers were exposed to 4%(w/v) of ‘undigested(non-hydrolysed)-soluble' fraction of protein sources used as feed ingredients [soybean meal(SBM) and casein], or alternative protein sources(spray dried plasma protein, and yellow meal worm), or controls for 6 h prior to RNA isolation and transcriptomics. All protein sources altered expression of unique biological processes in the epithelial cells. Exposure of intestinal organoids to SBM downregulated expression of retinol and retinoid metabolic processes as well as cholesterol and lipid biosynthetic pathways, consistent with the reported hypotriglyceridaemic effect of soy protein in vivo. These findings support the use of intestinal organoids as models to evaluate complex interactions between dietary ingredients and the intestinal epithelium and highlights some unique host effects of alternative protein sources in animal feed and potentially human food.
