Food-derived protein hydrolysates and peptides:anxiolytic and antidepressant activities,characteristics,and mechanisms

Globally,the prevalence of anxiety and depression has reached epidemic proportions.Food-derived protein hydrolysates and peptides delivered through dietary supplementation can avoid the negative risks associated with traditional pharmaceuticals while delivering superior anxiolytic and antidepressant effects.This review summarizes current research on food-derived anxiolytic and antidepressant protein hydrolysates and peptides,and subsequently analyses their physicochemical characteristics and elaborates on their mechanisms.The aim of this work is to contribute to the in-depth study and provide a theoretical foundation for the development of related products to better serve patients with anxiety and depression.

Preparation,characterization and antioxidant activity analysis of three Maillard glycosylated bone collagen hydrolysates from chicken,porcine and bovine

Bone collagen hydrolysates(peptides)derived from byproduct of animal product processing have been used to produce commercially valuable products due to their potential antioxidant activity.Maillard glycosylated reaction is considered as a promising method to enhance the antioxidant activity of peptides.Hence,this research aims at investigating the Maillard glycosylation activity and antioxidant activity of bone collagen hydrolysates from different sources.In this study,3 glycosylated bone collagen hydrolysates were prepared and characterized,and cytotoxicity and antioxidant activity were analyzed and evaluated.The free amino groups loss,browning intensity,and fluorescence intensity of G-Cbcp(glycosylated chicken bone collagen hydrolysates(peptides))were the heaviest,followed by G-Pbcp(glycosylated porcine bone collagen hydrolysates(peptides))and G-Bbcp(glycosylated bovine bone collagen hydrolysates(peptides)).The results of amino acid analysis showed that amino acid composition of different bone collagen hydrolysates was significantly different and the amino acid decreased to different degrees after Maillard glycosylated reaction,which may lead to differences in Maillard glycosylated reaction activity.Furthermore,the 3 glycosylated hydrolysates showed no significant cytotoxicity.The results showed that glycosylation process significantly increased the antioxidant activity of bone collagen hydrolysates,and G-Cbcp showed the strongest antioxidant activity,followed by G-Pbcp and G-Bbcp.Therefore,compared with the bone collagen hydrolysates,3 glycosylated hydrolysates showed significant characteristic and structural changes,and higher antioxidant activity.

The efficiency and safety evaluation of hemoglobin hydrolysate as a non-heme iron fortifier

Hemoglobin hydrolysate is derived from the enzymatic degradation of hemoglobin.This work aimed to evaluate whether hemoglobin hydrolysate promotes the absorption of non-heme iron and the safety of absorbed iron in mice by analyzing the iron binding content,iron circulation,and liver homeostasis.We found that hemoglobin hydrolysate promoted the absorption of non-heme iron with high efficiency in duodenum by spontaneously binding non-heme iron during digestion,and increased hepatic iron content by up-regulating divalent metal transporter 1,zinc transporter 14,but hepatic iron content only increased at 3 weeks.Duodenal iron entered the blood through ferroportin without restriction at 3 weeks,and excessive iron entered the liver and then affected the hepatocyte membranes permeability and lipid synthesis through oxidative stress.With the prolongation of dietary intervention,the up-regulated hepcidin acted on the ferroportin to restrict excess iron from entering the blood,and then the hepatic homeostasis recovered.In addition,hemoglobin hydrolysate enhanced the hepatic antioxidant capacity.Taken together,hemoglobin hydrolysate has a strong ability to promote the absorption of non-heme iron in vivo,and the absorbed iron is relatively safe due to the regulation of hepcidin.

Antihypertensive effects of whey protein hydrolysate involve reshaping the gut microbiome in spontaneously hypertension rats

Novel angiotensin-converting enzyme(ACE)inhibitory peptides were identified from whey protein hydrolysates(WPH)in vitro in our previous study and the antihypertensive abilities of WPH in vivo were further investigated in the current study.Results indicated that WPH significantly inhibited the development of high blood pressure and tissue injuries caused by hypertension.WPH inhibited ACE activity(20.81%,P<0.01),and reduced renin concentration(P<0.05),thereby reducing systolic blood pressure(SBP)(12.63%,P<0.05)in spontaneously hypertensive rats.The increased Akkermansia,Bacteroides,and Lactobacillus abundance promoted high short chain fatty acid content in feces after WPH intervention.These changes jointly contributed to low blood pressure.The heart weight and cardiomyocyte injuries(hypertrophy and degeneration)were alleviated by WPH.The proteomic results revealed that 19 protein expressions in the heart mainly associated with the wingless/integrated(Wnt)signaling pathway and Apelin signaling pathway were altered after WPH supplementation.Notably,WPH alleviated serum oxidative stress,indicated by the decreased malondialdehyde content(P<0.01),enhanced total antioxidant capacity(P<0.01)and superoxide dismutase activity(P<0.01).The current study suggests that WPH exhibit promising antihypertensive abilities in vivo and could be a potential alternative for antihypertensive dietary supplements.

Yeast hydrolysate attenuates lipopolysaccharide-induced inflammatory responses and intestinal barrier damage in weaned piglets

Background Intestinal inflammation is the main risk factor causing intestinal barrier dysfunction and lipopolysaccharide(LPS)can trigger inflammatory responses in various eukaryotic species.Yeast hydrolysate(YH)possesses multibiological effects and is received remarkable attention as a functional ingredient for improving growth performance and promoting health in animals.However,there is still inconclusive on the protective effects of dietary YH supplementation on intestinal barrier of piglets.This study was conducted to investigate the attenuate effects of YH supplementation on inflammatory responses and intestinal barrier injury in piglets challenged with LPS.Methods Twenty-four piglets(with an average body weight of 7.42±0.34 kg)weaned at 21 days of age were randomly assigned to one of two dietary treatments(12 replications with one pig per pen):a basal diet or a basal diet containing YH(5 g/kg).On the 22nd d,6 piglets in each treatment were intraperitoneally injected with LPS at 150μg/kg BW,and the others were injected with the same amount of sterile normal saline.Four hours later,blood samples of each piglet were collected and then piglets were euthanized.Results Dietary YH supplementation increased average daily feed intake and average daily gain(P<0.01),decreased the ratio of feed intake to gain of piglets(P sponse,evidenced by the increase o=0.048).Lipopolysaccharide(LPS)injection induced systemic inflammatory ref serum concentrations of haptoglobin(HP),adrenocorticotropic hormone(ACTH),cortisol,and interleukin-1β(IL-1β).Furthermore,LPS challenge resulted in inflammatory intestinal damage,by up-regulation of the protein or mRNA abundances of tumor necrosis factor-α(TNF-α),IL-1β,toll-like receptors 4(TLR4)and phosphor-nuclear factor-κB-p65(p-NFκB-p65)(P<0.01),and down-regulation of the jejunal villus height,the protein and mRNA abundances of zonula occludens-1(ZO-1)and occludin(OCC;P<0.05)in jejunal mucosa.Dietary YH supplementation decreased the impaired effects of ACTH,cortisol,HP,IL-1βand diamine oxidase in serum(P<0.05).Moreover,YH supplementation also up-regulated the jejunal villus height,protein and mRNA abundances of ZO-1 and OCC(P<0.05),down-regulated the mRNA expressions of TNF-αand IL-1βand the protein abundances of TNF-α,IL-1β,TLR4 and p-NFκB-p65 in jejunal mucosa in LPS-challenged pigs(P<0.01).Conclusion Yeast hydrolysate could attenuate inflammatory response and intestinal barrier injury in weaned piglets challenged with LPS,which was associated with the inhibition of TLR4/NF-κB signaling pathway activation.

Supplementation with yak (Bos grunniens) bone collagen hydrolysate altered the structure of gut microbiota and elevated short-chain fatty acid production in mice

In this study, yak bone collagen hydrolysate(YBCH)was produced by mixed proteases and provided to standard-diet mice at a different dose(low dose(LD), medium dose(MD), and high dose(HD))to investigate its effects on the composition of gut microbiota and short-chain fatty acids(SCFA)production. It was found that YBCH was mainly composed of small molecular peptides whose molecular weight below 2 000 Da. Notably, supplementation with different doses of YBCH could significantly downregulate the ratio of Firmicutes to Bacteroidetes in the fecal microbiota. At the family level, the Lachnospiraceae abundance was significantly reduced in the YBCH gavage groups(mean reduction ratio 41.7 %, 35.2%, and 36.4% for LD, MD, and HD group, respectively). The predicted functions of gut microbes in the MD group were significantly increased at “lipid metabolism” and “glycan biosynthesis and metabolism”. Moreover, the SCFA production in the YBCH groups was elevated. Especially, the concentration of acetic acid, propionic acid, and butyric acid in the MD group was separately increased 79.7%, 89.2%, and 78.8% than that in the NC group. These results indicated that YBCH might be applied in the development of functional food for intestinal microecological regulation.

Analysis of Molecular Weight Distribution and Antioxidant Activities of Cirrhinus molitorella Skin Collagen Hydrolysates

[Objective] The aim of this work was to identify molecular weight （MW） distribution and antioxidant activity of fish skin col agen hydrolysates. [Method] The MW distribution of hydrolysates was determined using both size exclusion chromatography and matrix-assisted laser desorption ionization time-of-flight mass spec-trometry （MALDI-TOF-MS）. Fish skin were treated by the alkaline protease 2709. [Result] The optional conditions for hyerolysis were time 3 h, temperature 55 ℃, pH 10.0, substrate concentration 80 g/L and E/S 4%. The results of both methods indi-cated that the molecular weight of col agen hydrolysates was from 400 to 1 800 Da, and the peptides’ molecular weight was less than 1 400 Da mostly. The reducing power and antioxidant/radical scavenging activity [1, 1-diphenyl-2-picryl-hydrazyl （DPPH） free radical scavenging activity] were determined. [Conclusion] The results reveal that the fishskin hydrolysate is a potential source of antioxidants.

Calcium-binding ability of soy protein hydrolysates

This present study investigated the ability of various soy protein hydrolysates (SPHs) in binding calcium. It was demonstrated that the amount of Ca-bound depended greatly on the SPHs obtained using different proteases, which included: neutrase, flavourzyme, protease M and pepsin. The maximum level of Ca-bound (66.9 mg/g) occurred when protease M was used to hydrolyze soy protein. Peptide fragments exhibiting high Ca-binding capacity had molecular weights of either 14.4 or 8–9 kDa. The level of Ca-bound increased linearly with the increment of carboxyl content in SPHs, and further deamidation on SPHs from protease M improved Ca-binding of the hydrolysate.

Antioxidant and Anti-aging Activities of Silybum Marianum Protein Hydrolysate in Mice Treated with D-galactose

Objective In the present study, we investigated the antioxidant and anti‐aging effects of Silybum marianum protein hydrolysate（SMPH） in D‐galactose‐treated mice. Methods D‐galactose（500 mg/kg body weight） was intraperitoneally injected daily for 7 weeks to accelerate aging, and SMPH（400, 800, 1,200 mg/kg body weight, respectively） was simultaneously administered orally. The antioxidant and anti‐aging effects of SMPH in the liver and brain were measured by biochemical assays. Transmission electron microscopy（TEM） was performed to study the ultrastructure of liver mitochondria. Results SMPH decreased triglyceride and cholesterol levels in the D‐galactose‐treated mice. It significantly elevated the activities of superoxide dismutase（SOD） and glutathione peroxidase（GSH‐Px）, and total antioxidant capacity（T‐AOC）, which were suppressed by D‐galactose. Monoamine oxidase（MAO） and malondialdehyde（MDA） levels as well as the concentrations of caspase‐3 and 8‐OHd G in the liver and brain were significantly reduced by SMPH. Moreover, it increased Bcl‐2 levels in the liver and brain. Furthermore, SMPH significantly attenuated D‐galactose‐induced liver mitochondrial dysfunction by improving the activities of Na＋‐K＋‐ATPase and Ca2＋‐Mg2＋‐ATPase as well as mitochondrial membrane potential（ΔΨm） and fluidity. TEM showed that the degree of liver mitochondrial damage was significantly decreased by SMPH. Conclusion The results indicated that SMPH protects against D‐galactose‐induced accelerated aging in mice through its antioxidant and anti‐aging activities.

A new method for determination of antithrombotic activity of egg white protein hydrolysate by microplate reader

A new method for the determination of antithrombotic activity of egg white protein hydrolysate （EWPH） was developed using a microplate reader. Reaction was carried out at 37℃and pH 7.2 with fibrinogen concentration 0.1%. Microplate reading was conducted at 405 nm. Inhibition rate of EWPH on thrombin activity showed linearity （R2 = 0.9971）, when the inhibition rate was in the range of 10-90%. The lower limit of detection （LLD, at 99.7% probability） and the biological limit of detection （BLD, at 99.7% probability） of the method were 10.643 and 40 mg/mL, respectively. The repeatability standard deviation （R.S.D.） was 1.08%. The standard deviation of the method was ±0.027 AT-U.

Low-protein diets supplemented with casein hydrolysate favor the microbiota and enhance the mucosal humoral immunity in the colon of pigs

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.

A new panaxadiol from the acid hydrolysate of Panax ginseng

A new panaxadiol （compound 1） was obtained from the acid hydrolysate of the total ginsenosides of Panax ginseng C. A. Meyer （Araliaceae）. On the basis of spectroscopic data and single-crystal X-ray diffraction data, its chemical structure was elucidated to be dammar-（E）-20（22）-ene-3β,12β,25 -tfiol.

Study on Enzymatic Hydrolysis of Gadus morrhua Skin Collagen and Molecular Weight Distribution of Hydrolysates

Process parameters on enzymatic hydrolysis and molecular weight （MW） distribution of collagen hydrolysates from Gadus morrhua skin were investigated. The optimal process parameters were obtained by the single-factor and orthogonal experiments. The molecular weight distribution of hydrolysates was determined using both Sephadex G25 partition and high speed liquid chromatography electricity spray mass spectrum （HPLC-ESI-MS）. Collagen hydrolysates were first gained by an alkaline protease ＂alcalase＂ for 3 h at temperature （50~C）, pH （10.0）, substrate concentration （75 g L-~）, and E/S （3%）. The molecular weight distribution of collagen hydrolysates ranged from 300 to 1 500 Da, and most of peptides were under 1 200 Da. Sephadex G25 partition and HPLC-ESI-MS should be successfully employed to determine the molecular weight distribution of collagen hydrolysates.

Identification of Odor Volatile Compounds and Deodorization ofPaphia undulata Enzymatic Hydrolysate

Unfavorable fishy odour is an inevitable problem in aquatic products. In the present study, headspace solid-phase mi- croextraction gas chromatography mass spectrometry （HS-SPME-GC-MS） analysis of volatiles from untreated samples and three deodorized samples （under the optimal conditions） ofPaphia undulata enzymatic hydrolysate revealed that the compounds contrib- uting to the distinctive odor were 1-octen-3-ol, n-hexanal, n-heptanal, 2,4-heptadienal, and 2,4-decadienal, whereas n-pentanal, n-octanal, n-octanol, benzaldehyde, 2-ethylfuran and 2-pentylfuran were the main contributors to the aromatic flavor. The deodoriz- ing effects of activated carbon （AC） adsorption, yeast extract （YE） masking and tea polyphenol （TP） treatment on a P. undulata en- zymatic hydrolysate were investigated using orthogonal experiments with sensory evaluation as the index. The following optimized deodorization conditions were obtained： AC adsorption （35 mg mL-1, 80℃, 40 rain）, YE masking （7 mgmL l, 45 ℃, 30 min） and TP treatment （0.4mgmL-l, 40℃, 50min）. AC adsorption effectively removed off-flavor volatile aldehydes and ketones. YE masking modified the odor profile by increasing the relative contents of aromatic compounds and decreasing the relative contents of aldehydes and ketones. The TP treatment was not effective in reducing the odor score, but it significantly reduced the relative content of alde- hydes while increasing that of alkanes. It is also notable that TP effectively suppressed trimethylamine （TMA） formation in a P. un- dulate hydrolysate solution for a period of 72 h.

Protein hydrolysates in animal nutrition：Industrial production, bioactive peptides,and functional significance

Recent years have witnessed growing interest in the role of peptides in animal nutrition. Chemical, enzymatic, or microbial hydrolysis of proteins in animal by-products or plant-source feedstuffs before feeding is an attractive means of generating high-quality small or large peptides that have both nutritional and physiological or regulatory functions in livestock, poultry and fish. These peptides may also be formed from ingested proteins in the gastrointestinal tract, but the types of resultant peptides can vary greatly with the physiological conditions of the animals and the composition of the diets. In the small intestine, large peptides are hydrolyzed to small peptides,which are absorbed into enterocytes faster than free amino acids（AAs） to provide a more balanced pattern of AAs in the blood circulation. Some peptides of plant or animal sources also have antimicrobial, antioxidant,antihypertensive, and immunomodulatory activities. Those peptides which confer biological functions beyond their nutritional value are called bioactive peptides. They are usually 2–20 AA residues in length but may consist of 〉20AA residues. Inclusion of some（e.g. 2–8%） animal-protein hydrolysates（e.g., porcine intestine, porcine mucosa,salmon viscera, or poultry tissue hydrolysates） or soybean protein hydrolysates in practical corn-and soybean mealbased diets can ensure desirable rates of growth performance and feed efficiency in weanling pigs, young calves,post-hatching poultry, and fish. Thus, protein hydrolysates hold promise in optimizing the nutrition of domestic and companion animals, as well as their health（particularly gut health） and well-being.

Production of Ammonium Lactate by Fed-batch Fermentation of Rhizopus oryzaefrom Corncob Hydrolysate

L-(+)-Lactic acid production from corncob hydrolysate as a cheap carbohydrate source by fed-batch fermentation of Rhizopus oryzaeHZS6 was studied. After 96 h of fermentation in a 5 L fermentor, the final concentration of ammonium L-(+)-lactate, average productivity(based on initial xylose concentration) and maximum dry cell weight were 132.4 g/L, 1.38 g/(L·h), and 8.9 g/L respectively. The optical purity of L-(+)-lactate was 98.8%.

Antihypertensive Properties on Spontaneously Hypertensive Rats of Peptide Hydrolysates from Silkworm Pupae Protein

Peptide hydrolysates of silkworm pupae protein with molecular weight of less than 5000 Da were prepared by ultrafiltration. The extracted peptide hydrolysates of silkworm pupae protein had inhibitory action on angiotensin-I-converting enzyme activity in vitro. The hydrolysates were orally administered to spontaneously hypertensive rats (SHR) in one period and long-term (four weeks). The results showed that the systolic blood pressure (SBP) of the treatment groups decreased in a dose-related manner. After one oral administration of silkworm protein hydrolysates with doses of 60, 20 and 5 mg/kg, the SBP of SHR decreased by 21.5, 13.8, and 9.0 mmHg in 1.5 h. After four weeks of the treatment in 80 mg/kg, the SBP decreased by 25 mmHg, with the antihypertensive activity close to 4 mg/kg of captopril;the SBP of the 40 mg/kg dose group also decreased by 17.5 mmHg. The peptide hydrolysate did not affect the SBP in normal, non-hypertensive rats in one period and long-term treatments. The acute toxicity research showed that the peptide hydrolysates were safe and without side effects. This research would be helpful in exploring the silkworm protein peptides as functional components for the antihypertension treatment.

Antioxidation Activities of Low-Molecular-Weight Gelatin Hydrolysate Isolated from the Sea Cucumber Stichopus japonicus

Gelatin extracted from the body wall of the sea cucumber （Stichopus japonicus） was hydrolyzed with flavourzyme. Low-molecular-weight gelatin hydrolysate （LMW-GH） of 700-- 1700 Da was produced using an ultrafiltration membrane bioreaetor system. Chemiluminescence analysis revealed that LMW-GH scavenges high free radicals in a concentration-dependent manner; IC50 value for superoxide and hydroxyl radicals was 442 and 285 μgmL-1, respectively. LMW-GH exhibited excellent inhibitory characteristics against melanin synthesis and tyrosinase activity in B16 cells. Furthermore, LMW-GH notably increased in- traeellular glutathione （GSH）, which in turn suppressed melanogenesis. LMW-GH performs antioxidation activity, holding the potential of being used as a valuable ingredient in function foods, cosmetics and pharmaceuticals or nutriceuticals.

Nutritional evaluation of caseins and whey proteins and their hydrolysates from Protamex

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.

Amino Acid Composition and Functional Properties of Giant Red Sea Cucumber (Parastichopus californicus) Collagen Hydrolysates

Giant red sea cucumber (Parastichopus californicus) is an under-utilized species due to its high tendency to autolysis.The aim of this study was to evaluate the functional properties of collagen hydrolysates from this species.The degree of hydrolysis (DH),amino acid composition,SDS-PAGE,emulsion activity index (EAI),emulsion stability index (ESI),foam expansion (FE),and foam stability (FS) of hydrolysates were investigated.The effects of pH on the EAI,ESI FE and FS of hydrolysates were also inves-tigated.The results indicated that the β and α1 chains of the collagen were effectively hydrolyzed by trypsin at 50℃ with an En-zyme/Substrate (E/S) ration of 1:20 (w:w).The DH of collagen was up to 17.3% after 3 h hydrolysis with trypsin.The hydrolysates had a molecular weight distribution of 1.1 17 kDa,and were abundant in glycine (Gly),proline (Pro),glutamic acid (Glu),alanine (Ala) and hydroxyproline (Hyp) residues.The hydrolysates were fractionated into three fractions (< 3 kDa,3 10 kDa,and > 10 kDa),and the fraction of 3 10 kDa exhibited a higher EAI value than the fraction of > 10 kDa (P<0.05).The fraction of > 10 kDa had higher FE and FS values than other fractions (P<0.05).The pH had an important effect on the EAI,ESI,FE and FS.All the fractions showed undesirable emulsion and forming properties at pH 4.0.Under pH 7.0 and pH 10.0,the 3 10 kDa fraction showed higher EAI value and the fraction of > 10 kDa showed higher FE value,respectively.They are hoped to be utilized as functional ingredients in food and nutraceutical industries.