Isolation and Characterization of Collagen and Antioxidant Collagen Peptides from Scales of Croceine Croaker (Pseudosciaena crocea)

Acid soluble collagen (ASC) from scales of croceine croaker (ASC-C) was successfully isolated with the yield of 0.37% ± 0.08% (dry weight basis), and characterized as type I collagen on the basis of amino acid analysis and electrophoretic pattern. The antioxidant hydrolysate of ASC-C (ACH) was prepared through a two-stage in vitro digestion (4-h trypsin followed by 4-h pepsin), and three antioxidant peptides (ACH-P1, ACH-P2, and ACH-P3) were further isolated from ACH using ultrafiltration, gel chromatography, and RP-HPLC, and their amino acid sequences were identified as GFRGTIGLVG (ACH-P1), GPAGPAG (ACH-P2), and GFPSG (ACH-P3). ACH-P1, ACH-P2, and ACH-P3 showed good scavenging activities on hydroxyl radical (IC₅₀ 0.293, 0.240, and 0.107 mg/mL, respectively), DPPH radical (IC₅₀ 1.271, 0.675, and 0.283 mg/mL, respectively), superoxide radical (IC₅₀ 0.463, 0.099, and 0.151 mg/mL, respectively), and ABTS radical (IC₅₀ 0.421, 0.309, and 0.210 mg/mL, respectively). ACH-P3 was also effectively against lipid peroxidation in the model system. The antioxidant activities of three collagen peptides were due to the presence of hydrophobic amino acid residues within the peptide sequences. The collagen peptides might be used as antioxidant for the therapy of diseases associated with oxidative stress, or reducing oxidative changes during storage.     

Influence of Amino Acid Compositions and Peptide Profiles on Antioxidant Capacities of Two Protein Hydrolysates from Skipjack Tuna (Katsuwonus pelamis) Dark Muscle

Influence of amino acid compositions and peptide profiles on antioxidant capacities of two protein hydrolysates from skipjack tuna (Katsuwonus pelamis) dark muscle was investigated. Dark muscles from skipjack tuna were hydrolyzed using five separate proteases, including pepsin, trypsin, Neutrase, papain and Alcalase. Two hydrolysates, ATH and NTH, prepared using Alcalase and Neutrase, respectively, showed the strongest antioxidant capacities and were further fractionated using ultrafiltration and gel filtration chromatography. Two fractions, Fr.A3 and Fr.B2, isolated from ATH and NTH, respectively, showed strong radical scavenging activities toward 2,2-diphenyl-1-picrylhydrazyl radicals (EC₅₀ 1.08% ± 0.08% and 0.98% ± 0.07%), hydroxyl radicals (EC₅₀ 0.22% ± 0.03% and 0.48% ± 0.05%), and superoxide anion radicals (EC₅₀ 1.31% ± 0.11% and 1.56% ± 1.03%) and effectively inhibited lipid peroxidation. Eighteen peptides from Fr.A3 and 13 peptides from Fr.B2 were isolated by reversed-phase high performance liquid chromatography, and their amino acid sequences were determined. The elevated antioxidant activity of Fr.A3 might be due to its high content of hydrophobic and aromatic amino acid residues (181.1 and 469.9 residues/1000 residues, respectively), small molecular sizes (3–6 peptides), low molecular weights (524.78 kDa), and amino acid sequences (antioxidant score 6.11). This study confirmed that a smaller molecular size, the presence of hydrophobic and aromatic amino acid residues, and the amino acid sequences were the key factors that determined the antioxidant activities of the proteins, hydrolysates and peptides. The results also demonstrated that the derived hydrolysates and fractions from skipjack tuna (K. pelamis) dark muscles could prevent oxidative reactions and might be useful for food preservation and medicinal purposes.     

Antioxidant and Antihypertensive Potential of Protein Fractions from Flour and Milk Substitutes from Canary Seeds (<Emphasis Type="Italic">Phalaris canariensis</Emphasis> L.)

Canary seed (Phalaris canariensis) is used to feed birds but it has been recently considered a promising cereal with nutraceutical potential for humans. The aim of this work was to analyze the protein fractions from canary seed flour and from milk substitutes (prepared by soaking the seeds in water 12 and 24 h), and to evaluate antioxidant and antihypertensive capacity of peptides obtained after in vitro digestion. Prolamins were the major protein fraction, followed by glutelins. After digestion, albumins and prolamins fractions from milks presented higher levels of peptides than flour, globulins showed more peptides in flour and glutelins were found in similar concentrations in all samples; 24 h milk prolamins had the highest concentration of peptides. Purification by high performance liquid chromatography (HPLC), sequencing of peptides, in vitro antioxidant ABTS (2,2′-azino-bis, 3-ethylbenzothiazoline-6-sulphonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays, and antihypertensive capacity (angiotensin converting enzyme (ACE) assay), indicated that peptides from canary seed prolamins were the most efficient compounds with antioxidant and antihypertensive activity. Canary seeds may be considered an accessible and cheap source to prepare milk substitutes with high contents of bioactive peptides with remarkable functional properties to promote better human health and healthy ageing.     

Purification of Antioxidant Peptides by High Resolution Mass Spectrometry from Simulated Gastrointestinal Digestion Hydrolysates of Alaska Pollock (Theragra chalcogramma) Skin Collagen

In this study, the stable collagen hydrolysate was prepared by alcalase hydrolysis and twice simulated gastrointestinal digestion from Alaska pollock skin. The characteristics of hydrolysates and antioxidant activities in vitro, including 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS^•+) scavenging activity, ferric-reducing antioxidant power (FRAP) and hydroxyl radical (OH·) scavenging activity, were determined. After twice simulated gastrointestinal digestion of skin collagen (SGI-2), the degree of hydrolysis (DH) reached 26.17%. The main molecular weight fractions of SGI-2 were 1026.26 and 640.53 Da, accounting for 59.49% and 18.34%, respectively. Amino acid composition analysis showed that SGI-2 had high content of total hydrophobic amino acid (307.98/1000). With the simulated gastrointestinal digestion progressing, the antioxidant activities increased significantly (p < 0.05). SGI-2 was further purified by gel filtration chromatography, ion exchange chromatography and high performance liquid chromatography, and the A_1a3c–p fraction with high hydroxyl radical scavenging activity (IC50 = 7.63 μg/mL) was obtained. The molecular weights and amino acid sequences of key peptides of A_1a3c–p were analyzed using high resolution mass spectrometry (LC-ESI-LTQ-Orbitrap-MS) combined with de novo software and UniProt of MaxQuant software. Four peptides were identified from A_1a3c–p, including YGCC (444.1137 Da) and DSSCSG (554.1642 Da) identified by de novo software and NNAEYYK (900.3978 Da) and PAGNVR (612.3344 Da) identified by UniProt of MaxQuant software. The molecular weights and amino acid sequences of four peptides were in accordance with the features of antioxidant peptides. The results indicated that different peptides were identified by different data analysis software according to spectrometry mass data. Considering the complexity of LC-ESI-LTQ-Orbitrap-MS, it was necessary to use the different methods to identify the key peptides from protein hydrolysates.     

Effects of Roasting on Phenolic Composition and In vitro Antioxidant Capacity of Australian Grown Faba Beans (Vicia faba L.)

Faba bean phenolic compounds encompassed phenolic acids, flavonols, proanthocyanidins and anthocyanins. Roasting faba beans for 120 min decreased the total phenolic, flavonoid and proanthocyanidin contents by 42, 42 and 30 %, respectively. Roasting beans for 120 min decreased the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, total equivalent antioxidant capacity and ferric reducing antioxidant power by 48, 15 and 8 %, respectively. High performance liquid chromatography-post column derivatisation revealed the generation of new phenolic compounds as a result of roasting. Antioxidant mechanism of bean less-polar phenolic compounds was largely based on free radical scavenging activity. The bean phenolic compounds with reducing capability were heat stable. Roasted faba bean extracts (70 % acetone, v/v) were fractionated into relatively polar and non-polar fractions; the latter contributed the majority of the antioxidant capacity. The extracts from beans with different seed coat colours differed in their phenolic compositions, which suggest different levels of potential benefits to health. Although roasting initially lowers the bean antioxidant capacity, prolonged roasting at 150 °C for 60 min and longer causes generation of new phenolic compounds and an increased antioxidant capacity. The findings encourage a wider ultilisation of faba beans for human foods particularly in baked/roasted products.     

Preparation and bioactivity evaluation of low salt peptide from sunf lower seed meal

Sunflower seed meal peptide as one sort of bioactive peptide has intensively application prospects. However, preparation of low salt peptide from sunflower seed meal with high efficiency remains a challenge. In this study, single and compound proteases were optimized to hydrolyze protein. Results showed that hydrolysis at pH 7.0 by proteases resulted in ash content in the range of 5.66%-7.37% and small peptides. Among all hydrolysis processes, sequential hydrolysis of Alcalase with Flavourzyme and Alcalase with Protamex showed higher nitrogen recovery ratio (67.66% and 66.49%, respectively). Furthermore, biological activities of peptides were investigated by testing their ABTS (2,2-azinobis (3-ethylben-zothiazoline-6-sulfonic acid) diammonium salt) radical scavenging activity, DPPH (2,2-diphenyl-1-picrylhydrazil) radical scavenging activity and angiotensin converting enzyme (ACE) inhibitory activity. Peptide hydrolyzed by Alcalase with Papain presented the highest antioxidant activity, followed by Alcalase with Protamex, with ABTS scavenging rate as 63.01% and 31.75%, and DPPH scavenging rate as 56.04% and 28.06%, respectively. Synchronously, peptide hydrolyzed by Alcalase with Protamex and Alcalase with Alcalase had the highest ACE inhibitory activity (56.74%, 56.76%). In conclusion, hydrolysis by proteases Alcalase with Protamex at pH 7.0 was the most effective method for the preparation of low salt peptide from sunflower seed meal, which could be an alternative for anti-oxidants and anti-vasoconstrictor.     

苦丁茶冬青粗多糖的分离表征及其抗氧化活性研究

水提醇沉法分离制备苦丁茶冬青粗多糖样品,对其进行初步分离表征（包括总糖、糖醛酸、蛋白质、氨基酸含量和红外光谱分析）和体外抗氧化活性（清除DPPH·自由基、·OH自由基清除能力和还原能力）研究。结果表明,苦丁茶冬青粗多糖中总糖含量为30.67％、糖醛酸含量为12.72％、蛋白质含量为9.35％;含有16种氨基酸成分,总含量为7.72％,其中含有7种人体必需氨基酸,占氨基酸总量的40.41％;红外光谱显示该粗多糖样品中含有α-吡喃糖环结构;此外,苦丁茶冬青粗多糖具有较强的体外抗氧化活性,且其抗氧化活性与多糖浓度之间存在良好的量效关系。     

Composition of Antioxidants and Amino Acids in Stevia Leaf Infusions

Stevia, a non-caloric natural sweetener with beneficial properties and considerable antioxidants and amino acids, is increasingly consumed as an infusion. This work evaluates the influence of the conditions (temperature: 50, 70 or 90 °C and time: 1, 5, 20 or 40 min) applied to obtain Stevia infusions, on antioxidants (total phenols, flavonoids and antioxidant activity) and amino acids. The total concentration of the eleven amino acids found was 11.70 mg/g in dried leaves and from 6.84 to 9.11 mg/g per gram of Stevia in infusions. However, infusions showed higher levels of certain amino acids (alanine, asparagine, leucine and proline), and greater values of the three antioxidant parameters in comparison with dry leaves. Temperature had more influence (minimum values at 50 °C and maximum at 90 °C) than time in the case of antioxidants. At 90 °C there were no important increases in the extraction of antioxidant compounds after 5 min; each gram of Stevia had 117 mg trolox (total antioxidant activity), 90 mg gallic acid (total phenols) and 56 mg catechin equivalents (flavonoids). Varying the temperature and time conditions no notable differences were observed in the concentrations of the majority of amino acids. However, the infusion treatment at 90 °C for 5 min was the best, as it gave the highest yield of 8 of the 11 amino acids. Therefore, with respect to the compounds analyzed in this study, the best way to obtain Stevia leaf infusions is the same as the domestic process, almost boiling water for a short time.     

The Effect of Thermal and Ultrasonic Treatment on Amino Acid Composition, Radical Scavenging and Reducing Potential of Hydrolysates Obtained from Simulated Gastrointestinal Digestion of Cowpea Proteins

The effect of thermal and ultrasonic treatment of cowpea proteins (CP) on amino acid composition, radical scavenging and reducing potential of hydrolysates (CPH) obtained from in vitro simulated gastrointestinal digestion of CP was evaluated. Hydrolysis of native and treated CP with gastrointestinal pepsin and pancreatin yielded CPH that displayed antioxidant activities based on oxygen radical scavenging capacity (ORAC), ferric reducing antioxidant power (FRAP) and superoxide radical scavenging activity (SRSA). CPH derived from the treated CP yielded higher ORAC values than CPH from untreated proteins. However, lower significant FRAP and SRSA values were observed for these samples compared to untreated CPH (p?<?0.05). Amino acid analysis indicated that CP processing decreased total sulphur-containing amino acids in the hydrolysates, particularly cysteine. The amount of cysteine appeared to be positively related to FRAP and SRSA values of CPH samples, but not ORAC. The results indicated that thermal and ultrasonic processing of CP can reduce the radical scavenging and reducing potential of the enzymatic hydrolysates possibly due to the decreased amounts of cysteine. Since the hydrolysates were generated with gastrointestinal enzymes, it is possible that the resulting compounds are produced to exert some health functions during normal consumption of cowpea.     

Polyphenolic Composition and Antioxidant Characteristics of Kumquat (<Emphasis Type="Italic">Fortunella margarita</Emphasis>) Peel Fractions

The polyphenolic composition of two Fortunella margarita (Nagami kumquat) specimens from Greece and Egypt was investigated employing fractionation by solvent partition and liquid chromatography-mass spectrometry. The main groups of phenolics identified in the different fractions generated were C-glycosylated flavones, O-glycosylated flavones, C-glycosylated flavanones, O-glycosylated flavanones, flavonols, chalcones, phenolic acids and derivatives thereof. The antioxidant potency of the fractions was assessed using two representative in vitro tests, including antiradical activity and hydroxyl free radical scavenging activity. It was revealed that the ethyl acetate fractions from both specimens contained the higher polyphenol content and exhibited the better antioxidant characteristics. The results indicated that F. margarita peels may be regarded as a rich source of potentially bioactive polyphenols.     

Fractionation of tryptic gliadin hydrolysates based on proline levels

The central domain (CD) and terminal domains (TDs) of wheat gliadins contain high and low levels of the amino acid proline (Pro), respectively. The CD is rather hydrophilic while the TDs are rather hydrophobic and contain most of the ionisable amino acids, although present only in low levels. Therefore, peptides derived from the CD or TDs may strongly differ in their physico–chemical properties. Trypsin cleaves peptide bonds at lysine and arginine residues which are mainly present in the TDs and was used in the present study. Several fractionation methods were examined to isolate CD and TD related peptides from tryptic hydrolysates. Pro was used as a marker for CD and TD related peptides. Both semi-preparative reversed-phase HPLC separation and fractionation of the water-soluble peptides of the tryptic gliadin hydrolysates by graded ethanol precipitation resulted in peptide fractions with different Pro levels. Whereas the fractions precipitating up to 90% ethanol were Pro rich, a Pro poor fraction consisting of small peptides was soluble in 90% ethanol solution. The water insoluble peptides of the tryptic hydrolysates also showed a low Pro level. Ultrafiltration of the water-soluble peptides using a 5-k membrane resulted in small Pro poor peptides and larger Pro rich peptides.     

Physicochemical and Antioxidant Properties of Gelatin and Gelatin Hydrolysates Obtained from Extrusion-Pretreated Fish (Oreochromis sp.) Scales

Fish gelatin and its hydrolysates exhibit a variety of biological characteristics, which include antihypertensive and antioxidant properties. In this study, fish gelatins were extracted from extrusion-pretreated tilapia scales, and then subjected to analyses to determine the physicochemical properties and antioxidant activity of the extracted gelatins. Our findings indicate that TSG2 (preconditioned with 1.26% citric acid) possessed the greatest extraction yield, as well as higher antioxidant activities compared with the other extracted gelatins. Hence, TSG2 was subjected to further hydrolyzation using different proteases and ultrafiltration conditions, which yielded four gelatin hydrolysates: TSGH1, TSGH2, TSGH3, and TSGH4. The results showed that TSGH4 (Pepsin + Pancreatin and ultrafiltration < 3000 Da) had a higher yield and greater antioxidant activity in comparison with the other gelatin hydrolysates. As such, TSGH4 was subjected to further fractionation using a Superdex peptide column and two-stage reverse-phase column HPLC chromatography, yielding a subfraction TSGH4-6-2-b, which possessed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity compared with the other fractions. Further LC-ESI/MS/MS analysis of TSGH4-6-2-b suggested two novel peptides (GYDEY and EPGKSGEQGAPGEAGAP), which could have potential as naturally-occurring peptides with antioxidant properties. These promising results suggest that these antioxidant peptides could have applications in food products, nutraceuticals, and cosmetics.     

灌木与乔木晒青茶生化成分与抗氧化活性研究比较

在灌木茶树留养改造形成乔木茶树的过程中,出现了改造中的灌木茶树、乔木茶树以及改造后的乔木茶树3种类型的茶树。本文对不同季节的上述3类茶树加工的晒青茶叶的生化成分与抗氧化活性进行比较研究。应用分光光度法、高效液相色谱法测定晒青茶的各内含成分含量,发现除苏氨酸、精氨酸和γ-氨基丁酸外,乔木晒青茶的茶氨酸、谷氨酸等15种氨基酸含量与游离氨基酸总量均高于灌木晒青茶（P<0.05）,而乔木晒青茶的茶多酚、没食子酸、1,4,6-O-没食子酰基-β-D-葡萄糖、表没食子儿茶素没食子酸酯（EGCG）的含量显著低于灌木晒青茶（P<0.05）,感官审评表明灌木晒青茶滋味较乔木晒青茶苦涩;正交偏最小二乘法（OPLS-DA）分析表明,两类茶聚成不同簇,进一步证实了茶叶生化成分存在差异。春季样品中,除羟自由基清除能力无显著差异外,乔木晒青茶的总抗氧化能力、DPPH自由基和超氧阴离子清除能力均显著高于灌木晒青茶（P<0.05）;秋季样品中,乔木晒青茶的总抗氧化能力显著高于灌木晒青茶（P<0.05）,但乔木晒青茶的超氧阴离子清除率则显著低于灌木晒青茶（P<0.05）,而DPPH和羟自由基清除活性二者差异不显著（P>0.05）。综上,与灌木晒青茶比较,改造形成的乔木晒青茶的氨基酸含量提高,多酚含量降低,苦涩度降低,部分抗氧化活性指标提高,表明灌木茶树留养改造成乔木茶树,能够提高晒青茶品质。     

Chemical composition and nutritional potential of the tribal pulse (Bauhinia malabarica Roxb)

Proximate and mineral composition, seed protein fractions, amino acid composition, fatty acid profiles and antinutritional factors were analysed for the seeds of the tribal pulse,Bauhinia malabarica. The seeds contained higher amount of crude lipid when compared with most of the domesticated pulses. The seeds were rich in minerals like Ca, Mg and Fe. Glutelins (45%) constituted the major seed protein fraction followed by globulins which accounted for about 34%. Cystine and methionine were found to be the limiting essential amino acids; whereas tyrosine and phenylalanine content were fairly high when compared with WHO requirement pattern. The unsaturated fatty acids, oleic acid and linoleic acid, were predominant in seed lipids. Presence of antinutritional substances like total free phenols, tannins, L-DOPA and haemagglutinating activity also were analysed/assayed.     

椰肉中醇溶蛋白抗氧化活性

从脂质过氧化的抑制作用、对DPPH自由基(DPPH.)的清除作用、羟基自由基(·0H)的清除作用、Fe2+的络合能力等几个方面研究椰子醇溶蛋白的抗氧化活性,并以BHT、没食子酸,VE做为对照.结果表明,椰子醇溶蛋白对Fe2+具有很强的络合能力.对DPPH·有较强的清除能力,同时具有一定的清除·OH能力和还原能力,但其抑制脂质体过氧化作用的能力较弱.这表明椰子醇溶蛋白有较好的抗氧化活性,但其溶解性限制了其在脂溶性食品中的应用.     

辣木籽多糖的超高压辅助提取工艺及其抗氧化活性分析

超高压辅助提取技术是一种天然产物新型绿色高效提取技术,在保证提取效率的同时能最大限度保持天然产物的生物活性。为探究超高压辅助提取技术对辣木籽多糖的提取效果及其抗氧化活性的影响,本研究以辣木籽为原料,通过超高压辅助提取技术提取辣木籽中的水溶性多糖,以多糖得率为考察指标,以提取压力、提取时间、料液比、粉碎度作为单因素,在单因素试验基础上,采用正交试验设计方法优化辣木籽多糖的超高压辅助提取工艺,并通过测定总抗氧化能力、清除DPPH自由基（DPPH•）和羟自由基（•OH）的能力来分析辣木籽水溶性多糖的抗氧化活性。结果表明,辣木籽多糖最佳的超高压辅助提取工艺条件为：提取压力100 MPa、保压时间6 min、料液比（g/mL）1:15、粉碎度100目筛,在最佳提取工艺条件下辣木籽多糖最大提取得率为0.346%;在测试范围内辣木籽多糖清除DPPH自由基能力随多糖浓度的增加而增加,并有较好的线性关系,清除率达到50%时对应的浓度（IC₅₀）为0.0439 g/L,但是其抗氧化能力低于相同浓度的Vc,在测试范围内清除羟自由基能力也随辣木籽多糖浓度的增加而增加,也有较好的线性关系,IC₅₀为0.1666 g/L,其抗氧化能力与同浓度的Vc较接近,辣木籽多糖的总抗氧化能力为0.0482 mmol/L（以硫酸亚铁的当量浓度表示）。与热水提取方法相比,超高压辅助提取方法能够缩短提取时间,提取温度大大降低,提取效率显著提高;而且提取的辣木籽水溶性多糖具有较好的抗氧化能力,可以作为天然抗氧化剂进行开发利用。本研究结果可为天然抗氧化剂的开发、辣木资源的综合开发及高值化利用提供技术支持和参考。     

刺五加叶不同部位抗氧化活性的研究

目的研究刺五加叶不同部位在不同浓度下的体外抗氧化活性。方法采用清除ABTS+自由基、DPPH自由基评价法和测定还原性法,利用紫外分光光度计法测定刺五加叶的总提液、10%、30%和50%大孔吸附树脂醇洗脱部位在不同浓度下的抗氧化能力。结果刺五加叶不同部位抗氧化活性的强弱顺序为:30%部位>10%部位>总提液>50%部位,随着各部位浓度的增加,抗氧化能力增强。ABTS+自由基和DPPH自由基模型中,在浓度为0.64mg/mL时,30%的部位对ABTS+DPPH自由基模最大清除率分别达到99.541%、95.331%,与同浓度VC的清除率接近,其次浓度0.64mg/mL的10%部位的最大清除率分别达到96.324%、88.033%。在还原性法测定中30%部位在浓度为0.64mg/mL时,吸光度为2.269,与VC较接近,同浓度时10%部位的吸光度为2.155,次于30%部位。结论刺五加叶抗氧化能力较好,各种评定方法结合分析,其30%部位最好,可以开发为抗氧化剂,为进一步研究和开发刺五加叶提供了科学依据。     

Antithrombotic Activity of Brewers’ Spent Grain Peptides and their Effects on Blood Coagulation Pathways

Antithrombotic activity of brewers’ spent grain peptides before and after simulated gastrointestinal digestion and their effects on blood coagulation pathways were evaluated. Two hydrolysates were produced using sequential enzymatic systems: alkaline protease + Flavourzyme (AF) and neutral protease + Flavourzyme (PF). Simulation of gastrointestinal digestion of AF and PF hydrolysates was made using porcine pepsin and pancreatin enzymes, obtaining the corresponding digested samples: AFD and PFD, respectively. Peptides were fractionated by ultrafiltration using a 1 kDa cut-off membrane. Hydrolysates had peptides with medium and low molecular weights (2100 and 500 Da, respectively), and Glu, Asp, Leu, Ala, and Phe were the most abundant amino acids. Gastrointestinal digested hydrolysates presented high proportion of small peptides (~500 Da), and higher amount of Val, Tyr, and Phe than hydrolysates. Mass spectrum (HDMS Q-TOF) of AFD-ultrafiltered fraction <1 kDa exhibited peptides from 500 to 1000 Da, which are not present in AF. PFD showed the generation of new peptides from 430 to 1070 Da. All samples showed thrombin inhibitory activity. However, no effect was observed on prothrombin time. Peptides <1 kDa from hydrolysates and digested samples delayed thrombin and thromboplastin time respect to the control (~63%). Also the samples showed thrombin inhibitory activity at common pathway level. Thus, brewers’ spent grain peptides exerted their antithrombotic activity by inhibiting the intrinsic and common pathways of blood coagulation. This is the first report to demonstrate that brewers’ spent grain peptides are able to delay clotting time after simulated gastrointestinal digestion.     

Characterization of Amaranthus hypochondriacus seed protein fractions,and their antioxidant activity after hydrolysis with lactic acid bacteria

The objective of this work was to obtain and characterize five Amaranthus hypochondriacus seed protein fractions followed by the determination of antioxidant activities of hydrolysates produced with two Lactobacillus species. Molecular weights of polypeptides (SDS-PAGE) in fractions were 14–97 kDa and those identified by LC-MS/MS include agglutinin, trypsin inhibitor, and cystatin. Each fraction was hydrolyzed with Lactobacillus helveticus and Lactobacillus plantarum at their exponential (4 and 8 h) and stationary (14 and 24 h) growth phases. The free amino group content in hydrolysates produced with L. helveticus was up to 5.15% compared to 4.79% in those obtained with L. plantarum. In the antioxidant tests, higher ROO• scavenging activities were obtained for hydrolysates from albumins (1037 ± 80 μM TE/mL) and glutelins (1793 ± 168 μM TE/mL) fractions produced with L. helveticus and L. plantarum, respectively. Scavenging activities of •OH radicals were higher for 11S-G hydrolysates with both Lactobacillus at the stationary phase.     

超滤法分离花生肽及其抗氧化活性的研究

用碱性蛋白酶解法制备花生多肽,将得到的花生肽酶解液通过超滤的方法进行分离得到不同分子量段的多肽,对其进行抗氧化活性的研究得到抗氧化活性最强的分子量段的多肽。将花生短肽酶解液稀释并分别通过5KD、3KD的卷式纤维膜进行超滤得到三个分子量段的花生短肽。比较各分子量段的多肽对二苯代苦味酰基（DPPH）自由基的清除率、抗亚油酸氧化能力、羟自由基的清除率及还原力。结果表明：花生肽具有一定的抗氧化能力,且3KD以下的多肽抗氧化能力最强,3-5KD分子量段的多肽次之,大于5KD分子量的多肽表现出较弱的抗氧化能力。