Amino acid composition of non-absorbed peptide fractions in rat small intestines. III. Feeding of wheat gluten and molasses yeast. Essential amino acid content of feces]

The authors investigated the nitrogen content of different fractions and in particular the aminoacid composition of the trichloroacetic acid (TCA)-precipitable and TCA-soluble peptides in the distal small intestine of rats which had been fed experimental diets (nitrogen-free or containing casein, wheat gluten or molasses yeast as a protein carrier). Parallel with this, they determined in growing rats the nitrogen and amino-acid digestibility of molasses yeast and wheat gluten according to the technique of faecal analysis, and studied the amino-acid composition of the faecal protein. Independently of the dietary protein fed, the peptide fractions of the content of the intestine showed an amino-acid composition that was largely similar to that found in case of nitrogen-free diet. The amino-acid composition of the faecal protein is, independently of the dietary protein given, relatively constant and corresponds approximately to the intestinal loss protein. There is agreement between the amino-acid composition of the TCA-precipitable peptide fractions and those of the corresponding faecal proteins. These components seem to be of endogenous origin.     

The intestinal phase of peptide absorption

Peptides and not amino acids are the prevailing degradation products of protein digestion which are formed in the intestinal lumen and are absorbed from the mucosa. These two families differ in absorption. The differences become manifest when the absorption of peptide mixtures is compared with that of equimolecular mixtures of free amino acids. The absorption of peptides occurs in two different ways: 1. Transport of intact peptides through the membrane into the mucosal cell and subsequent hydrolysis by intracellular peptide hydrolases. 2. Hydrolysis of the peptides by peptide hydrolases localized on the luminal side of the mucosal cell membrane and subsequent transport of the amino acids thus formed through the membrane. The two mechanisms of absorption do not exclude each other. The way by which energy is supplied for the transport is not yet elucidated. The transport of intact peptides is of nutritive importance only in case of dipeptides and tripeptides. It enables in particular the introduction of peptides that cannot be cleft by membrane-bound peptide hydrolases. The hydrolysis of peptides by membrane-bound peptide hydrolases and the subsequent transport of released amino acids is of importance for long-chain peptides. The difference in absorbing behaviour between the free amino acids released in the intestinal lumen and the amino acids released by peptide hydrolases at the mucosal membrane is discussed.     

Comparative studies on the enzymatic absorption of protein hydrolysates in the small intestine of the rat. 3. The absorption trypsins thermatatic and trypsin-thermatatic hydrolysates of a fava bean protein isolate compared to an equimolar mixture of free amino acids

Tryptic, thermitatic, and tryptic-thermitatic Faba bean protein hydrolyzates as well as their equimolar mixture of amino acids were perfused through proximal and distal parts of the intestine (10 cm length) of non-narcotized rats. The total amino-acid concentration of the perfused solution was 50 mM. The absorption of nitrogen and total amino acids from the tryptic and tryptic-thermitatic hydrolyzates was lower than that from the amino-acid mixture, the absorption from the thermitatic hydrolyzate was in accordance with that from the amino-acid mixture. The absorption pattern of the amino acids which preferably undergo a peptidic absorption is similar with the three hydrolyzates: in the proximal intestinal part this concerns glutamic acid and serine, in the distal intestinal part--methionine, alanine, glycin, and serine. The absorption pattern of the amino acids is different between the three hydrolyzates and the amino-acid mixture. Between the absorption pattern of the amino acids from the three hydrolyzates little differences were evident only in the proximal intestinal part. The coefficients of variation of the tryptic-thermitatic hydrolyzates are in accordance with those of the amino-acid mixture, whereas that of the thermitatic hydrolyzates is significantly lower. In the distal intestinal part all supplied forms are more rapidly absorbed than in the proximal part of the intestine.     

Importance of peptides from enzymatically degraded proteins as components of foodstuffs

In comparison with proteins, peptides occur only in very small amounts in foods. An exception are peptides, which result from proteins to a higher or lower extent in the course of natural or microbial disintegration or ripening processes. Recently, proteinases are used for process optimization or quality improvement in the production or processing of foods. Usually, the protein degradation in these cases is only small, but functional properties may be improved. Peptides frequently show a bitter taste and may limit the sensory value of foods. Sources and possibilities to remove the bitter taste are discussed. In dietetics protein-free peptide diets are introduced for special indications. They contain oligopeptides which are produced by intensive hydrolysis of proteins. The intestinal absorption of small peptides is different from that of free amino acids. Finally, physiological effects of some dietary peptides are discussed (e.g. peptides as enzyme inhibitors, coeliac disease as a gliadine-peptide induced enteropathy, exorphines as neurotransmitters).     

Bioactive peptides derived from milk proteins. Structural,physiological and analytical aspects

The primary function of dietary proteins is to supply the body adequately with indispensable amino acids and organic nitrogen. Little attention has been paid up to date to milk proteins, in particular caseins, that are currently the main source of biologically active peptides, although other animal as well as vegetable proteins are known to contain potentially bioactive sequences. Such regulatory peptides can be released by enzymatic proteolysis of caseins in vitro and in vivo and may act as potential physiological modulators of metabolism during the intestinal digestion of the diet. It has been proved that bioactive peptides derived from caseins, such as β-casomorphins and phosphopeptides, can be released during gastrointestinal passage. It is also evident that peptides originating from food proteins should be taken into account as potential modulators of various regulatory processes in the body. The possible regulatory effects concern nutrient uptake (phosphopeptides, casomorphins), postprandial hormone secretion (casomorphins), immune defense (immunopeptides, casokinins, casomorphins) and neuroendocrine information transfer (casokinins). The advances in the research field of bioactive peptides are driven by a molecular understanding of biological processes, and analytical techniques are a critical component of this understanding. Different up-to-date methods, including peptide synthesis and immunochemistry, have been applied to the chemical characterization of bioactive peptides. Especially casein derived peptides have already found interesting applications, both as dietary supplements (phosphopeptides) and as pharmaceutical preparations (phosphopeptides, β-casomorphins). The question of'what kinds of bioactive peptides are beneficial and desirable as food constituents or as drugs' should be always carefully examined. However, the possibilities for the design of dietary products and ‘natural’ drugs look promising.     

Comparative studies on the enzymatic absorption of protein hydrolysates in the small intestines of the rat. 3. The absorption of trypsin, thermitatic and trypsin thermitatic wheat gluten hydrolysates in comparison with equimolar mixtures of free amino acids

Tryptic, thermitatic, and tryptic-thermitatic wheat-gluten hydrolyzates as well as their equimolar amino-acid mixture were perfused through proximal and distal parts of the intestine (10 cm length) of non-narcotized rats. The total amino-acid concentration of the perfused solution was 50 mM. The tryptic hydrolyzate showed a significantly lower absorption of nitrogen and total amino acids than the amino-acid mixture. Both the supplied forms were very different as to their absorption pattern of the amino acids. The high variability of the percental absorption of the individual amino acids of the tryptic hydrolyzate results in a high coefficient of variation. The absorption of nitrogen and total amino acids from thermitatic and tryptic-thermitatic hydrolyzates is equal to that from the amino-acid mixture. In a peptidic form glutamic acid is more rapidly absorbed from the two hydrolyzates, and methionine from the tryptic-thermitatic hydrolyzate in both the intestinal parts. As to alanine and glycine this concerns only the distal intestinal part for both the hydrolyzates. There are no differences between the absorption patterns of the two hydrolyzates but in comparison with the amino-acid mixture and the tryptic hydrolyzate differences were evident. The coefficients of variation of both the hydrolyzates are significantly lower as compared to those of the tryptic hydrolyzate and the amino-acid mixture. All forms of supply are more rapidly absorbed in the distal than in the proximal part of the intestine.     

Comparative study of partial amino acid sequences of prolamins and glutelins from various cereals. VII. Amino acid sequences of prolamin peptides

The peptide fractions isolated from chymotryptic hydrolysates of wheat, rye and barley prolamines [this journal (1984) 178:173] were separated into pure peptides by high-performance liquid chromatography on octadecyl silica gel. The peptides which were most abundant were analyzed for amino acid composition and in part for amino acid sequence. Besides peptides which are typical for only one of the cereals investigated, peptides of similar composition were found in all three prolamines. These contain repeating sequences and are built up of mainly Gln (Q), Pro (P) and hydrophobic amino acids (X) such as Phe, Tyr, Ile, Val and Leu. One of the most frequent partial sequences is QQPQQPXP.     

Chia Seed (Salvia hispanica L.) as a Source of Proteins and Bioactive Peptides with Health Benefits: A Review

The consumption of chia seed (Salvia hispanica L.) has increased in recent years due its high content of omega‐3 fatty acids and dietary fiber. This seed also has a high concentration of proteins and essential amino acids, becoming a promising source of bioactive peptides. The objective of this review was to identify the composition and the beneficial effects of chia seeds (S. hispanica L.), their proteins, peptides, and their potential impact on human health. The UniProt database was used to identify the chia proteins and their amino acid sequences. The BIOPEP database was used to analyze the peptides's bioactive potential. A total of 20 proteins were cataloged in chia seed, 12 of those were involved in the regular metabolic processes of the plant cells. However, eight proteins were specifically related to production and storage of plant lipids, thus explaining the high concentration of lipids in chia seeds (around 30%), especially omega‐3 fatty acids (around 20%). The analyses of amino acid sequences showed peptides with bioactive potential, including dipeptidyl peptidase‐IV inhibitors, angiotensin‐converting enzyme inhibitors, and antioxidant capacity. These results correlated with the main health benefits of whole chia seed in humans such as antioxidant capacity, and hypotensive, hypoglycemic, and anticholesterolemic effects. Such relation can be associated with chia protein and peptide compositions and therefore needs further investigation in vitro and in vivo.     

Comparative study of the absorption of enzymatic protein hydrolysates in the small intestine of the rat. 2. The absorption of tryptic, thermitatic and tryptic-thermitatic casein hydrolysates compared with the equimolar composition of free amino acids

Tryptic, thermitatic, and tryptic-thermitatic casein hydrolyzates as well as their equimolar amino-acid mixture were perfused through proximal and distal parts of the intestine (10 cm length) of nonanaesthesized rats. The total amino-acid concentration of the perfused solution was 50 mM. The absorption of nitrogen and total amino acids respectively did not reveal significant absorption advantages in favour of the hydrolyzates. In contrast to this, some peptidic bound amino acids of these hydrolyzates show a significantly better absorption as compared to free amino acids. At this, dependences of the kind of hydrolyzate and the part of the intestine are evident. Glutamic acid, e.g. is generally more rapidly absorbed when peptidic bound; methionine is more rapidly absorbed only from the thermitatic and tryptic-thermitatic hydrolyzates, and alanine and glycin only in the distal part of the intestine. Independent of the amino acid or peptide substrate the total absorption of all the amino acids is higher in the distal part of the intestine. The comparison concerning the ranking order of the single amino-acid absorption rates shows in the two parts of the intestine distinct differences between the amino-acid mixtures and the enzymatic hydrolyzates. The lowest differences were found between the thermitatic and the tryptic-thermitatic hydrolyzate. Both of them have approximately the same degrees of hydrolysis (30 and 35%, respectively). The variability of the amino-acid absorption from the three casein hydrolyzates is lower in comparison with the amino-acid mixture. The tryptic-thermitatic hydrolyzates have the lowest coefficients of variability.     

The Gel Filtration Chromatographic‐Profiles of Proteins and Peptides of Wort and Beer: Effects of Processing — Malting,Mashing, Kettle Boiling,Fermentation and Filtering

Barley and malt proteins, of infusion (IoB) and decoction (EBC) mashing worts as well as commercial wort and beer, obtained from the Castlemaine Perkins brewery, Brisbane, were gel filtered, with or without further treatments. A general, similar pattern of protein and peptide profiles emerged from barley malt and beer. This confirmed the widely assumed fact that beer proteins descend from barley, some transformed and others perhaps mostly unchanged by processing. In the gel‐filtrate profiles, a maximum of 8 or 9 fractions were discerned. These fractions were collected and quantified for protein contents and amino acid compositions. The first four fractions contained the proteins and polypeptides of molecular weight higher than 14,000. Consequently, the remaining fractions contain the smaller peptides (<14,000), that were completely removed by dialysis. The effects of processing on proteins and peptides varied contingent upon the type of processing step considered and the pre‐chromatographic treatment. Malting was the most effective process remarkably increasing the soluble protein contents, especially the smaller peptide fractions and the colour development. This is the first report, as far as we are aware of, on the gel filtration profiles of wort and beer low molecular weight peptides including those of barley wort. The importance of the smaller peptides in foam formation and retention cannot be overemphasised. The amino acid composition of the fractions revealed much more diversity than was observed in the comparison of the profiles. Proline content of fraction 1 resembled that of barley soluble proteins while fractions F2, F3 and F4 that of glutelin and only fraction 8 that of hordein. The latter, suggests that hordeins or, at least the peptide products rich in proline, are likely to be completely digested to amino acids, during malting.     

Optimization of amino-acid profile of protein-peptide products used in preparation of functional beverages

The message is devoted to optimization of amino acid structure of protein/peptide composition for its use as a component (a key ingredient) of functional drinks of various purposes. A basis of a composition forms protein/peptide product (PPP) received by enzymatic hydrolysis of meat and bone residues of processing of a bird, representing rather unvaluable food raw material. Molecular-mass distribution of fractions in PPP is quantitatively determined by an exclusion chromatography method of average pressure with the subsequent integration of chromatograms with a weight method. It can be supported the essential decrease of initial antigenicity of processed food raw materials as almost 80% from all sum protein/peptides and peptide/amino acid fractions are submitted by amino acids and peptides which molecular weight < or = 6 kD. The analysis of amino acid structure of PPP shows that the increase of its biological value assumes the increase of its score on four irreplaceable amino acids: leucine, tryptophan, phenylalanine and tyrosine (totally). For getting of the protein/peptide compositions of high biological value, the "above enrichment" of PPP by total protein of the cow milk is carried out. The modeling of a process of getting a two-component mix is carried out by a graphic method. The least percentage of the added total protein of a cow milk to PPP, at which is reached the amino acid score of the mix equal to 1,0, makes 26% from weight of the mix. The subsequent publications will be devoted to an estimation of biological value of a key component of functional food in the form of instant drinks in experiments in vivo on laboratory animals.     

奇亚籽蛋白及其活性肽的研究进展

奇亚籽富含优质蛋白质,且包含8种必需氨基酸。奇亚籽蛋白具有乳化、发泡等许多优良的功能性质,其制备的活性肽具有抗氧化、抗高血压等生物性能。本文综述了近年来国内外有关奇亚籽蛋白的氨基酸组成、提取分离工艺、功能性质、生物活性肽的制备及其生理功能方面的研究,并对奇亚籽蛋白在未来的发展方向以及在食品领域的开发应用作出了展望。     

Bioavailability of peptides from casein hydrolysate in vitro: Amino acid compositions of peptides affect the antioxidant efficacy and resistance to intestinal peptidases

Measurements of in vitro bioactivity to support health benefits of bioactive compounds should be accomplished with estimates of their bioavailability to bolster nutritional significance to health claims. In vitro bioavailability of casein and casein peptides (casein hydrolysate and four peptide fractions) measured by the amount of peptide nitrogen is discussed. Antioxidant activities during gastrointestinal digestion and Caco-2 cell absorption were investigated as indices of peptide degradation. The antioxidant capacities of Trolox equivalent and oxygen radicals were used for assessing antioxidant efficacy of surviving peptides. Results showed that casein hydrolysate improved bioavailability compared to casein. Amino acid composition of peptides affected the resistance of peptides to digestive enzymes and intestinal peptidases. The acidic peptide fractions had higher bioavailability and a higher residual ratio of antioxidant activity. The peptides in the digest and absorbate of acidic fraction F1 with the highest bioavailability (23.14%) and the residual ratio of antioxidant activity were identified, and 12 intact, absorbed peptides (IAP) were obtained. Eleven of twelve of the IAPs were from β-Casein, and their amino acid components were rich in acidic and hydrophobic amino acids. Identification of IAPs might provide insight into the mechanism of how peptide structure provides resistance against peptidases by Caco-2 cells.     

Oxygen radical absorbance capacity of peptides from egg white protein ovotransferrin and their interaction with phytochemicals

Egg proteins are important components for the development of functional foods and nutraceuticals. This study investigated the oxygen radical-scavenging effects of egg white protein ovotransferrin (including the whole protein, thermolysin–pepsin hydrolysate, and its bioactive peptides) and their interactions with four phytochemicals (vitamin C, epigallocatechin gallate (EGCG), caffeic acid, and quercetin), based on the oxygen radical absorbance capacity (ORAC) assay. The results showed that the hydrolysate possessed significantly higher oxygen radical absorbance capacity than the whole protein. The peptide IRW possessed very strong oxygen radical-scavenging effect, which might be attributed to its constitutive amino acid tryptophan (W) and the peptide bond between arginine (R) and W. Both the hydrolysate and the whole protein of ovotransferrin showed negative effects on four phytochemicals. However, the interactions between peptides/amino acids and the four phytochemicals are complex: some are positive, while others are negative. The results suggest that not only the phytochemicals, but also the hydrolysates or the peptides from the egg proteins could be a potential source of natural antioxidants.     

乳清蛋白及其肽制品功能特性的研究

乳清蛋白富含必需氨基酸是膳食极好的氮源。另外,乳清蛋白和其肽制品还具有许多功能特性,这些产品还具有作为功能性食品及食品功能性配料的作用。本文简述了乳清蛋白及其肽制品功能特性与人类健康的关系。     

草鱼抗氧化肽的美拉德反应特性研究

本研究通过单独加热草鱼肽和添加木糖加热反应制备热降解产物（TDPs）和美拉德反应产物（MRPs),运用DPPH?抑制率、肽分子量分析及氨基酸分析等方法评价不同反应时间草鱼肽热降解产物及美拉德反应产物的抗氧化性能、分子量分布以及氨基酸组成的变化规律,深入探讨了反应时间对草鱼肽的美拉德反应产物特性的影响。研究发现,单独加热草鱼肽对各时间段热降解产物的总氨基酸含量无显著影响,游离氨基酸含量显著增加,而3000 Da以下肽段含量增多。美拉德反应体系样品随着反应时间的增加,产物中总氨基酸及游离氨基酸含量下降,其中精氨酸及赖氨酸减少尤其显著,分子量大于3000 Da的肽段明显增多,3000 Da以下的肽段含量降低,此外,美拉德反应产物中抗氧化活性显著增强。     

Partially Hydrolyzed Soy Protein Shows Enhanced Transport of Amino Acids Compared to Nonhydrolyzed Protein across an Intestinal Epithelial Cell Monolayer

Consumption of protein hydrolysates has been proposed to stimulate muscle anabolism more than intact (nonhydrolyzed) proteins via accelerated delivery of amino acids for muscle protein synthesis (MPS). We evaluated whether the rate of amino acid uptake and transport across intestinal cells was enhanced for soy protein hydrolysates versus nonhydrolyzed soy protein. Intact and partially hydrolyzed proteins were subject to simulated gut digestion and applied to the apical surface of Caco‐2 monolayers. Basolateral media was harvested after 3 h and quantitatively analyzed for free amino acids using ion‐exchange chromatography and comparison to an included reference standard. Basolateral concentrations of all amino acids were higher (mean 32%) for hydrolyzed versus nonhydrolyzed protein with the greatest differences in histidine, lysine, and valine. Scale‐up production of the soy protein hydrolysate did not diminish its enhanced absorption properties. These data support the hypothesis that hydrolyzed soy protein may provide dietary amino acids that are more rapidly transported across the intestinal epithelium versus intact soy protein. This would be important under conditions where rapid and increased levels of amino acids are needed such as in the stimulation of MPS.     

高F值活性肽的生物活性、制备技术及 吸收评价技术的研究进展

高F值活性肽(HFRP)是指一类由2~9个氨基酸残基所组成的小肽, 一般F值大于20。HFRP是一种重要活性肽, 具有抗疲劳, 补充人体必需氨基酸, 促进酒精代谢, 保肝护肝, 促进氮储留和蛋白质合成, 抑制蛋白质分解, 改善手术后和卧床病人的蛋白营养状况等多种生理功能, 因此具有良好开发应用前景。本文综述了高F值活性肽的生理功能及用途、原料及制备技术、耐消化性及肠吸收评价技术的研究进展, 为新型活性肽保健食品的研究开发提供理论依据和技术支持。     

Semiessential and nonessential amino acids in parenteral feeding]

With reference to a critical study of the relevant literature and to results from their own investigations, the authors emphasize the importance of the semi-essential and non-essential amino acids arginine, histidine, tyrosine, cystine and glutamic acid for the completion of essential amino-acid mixtures destined for parenteral feeding. MADDEN'S assumption that intravenously applied glutamic acid is intolerable was not supported by the authors' experiments in dogs. As evidenced by the increase of blood urea, the glutamic acid-containing amino-acid mixture was well utilizable. When glutamic acid is present, proline and alanine are obviously superfluous in amino-acid mixtures for parenteral feeding, since they are easily formed from glutamic acid by intermediary metabolism.     

大豆生物活性肽的分离及其抗氧化活性研究

为制各具有抗氧化活性的大豆生物活性肽,本实验采用酶解处理大豆蛋白粉,对酶解产物进行葡聚糖Sephadex G-25凝胶柱层析分离,并对其各组分分子量分布及其抗氧化性进行了研究,对抗氧化活性最佳的肽段进行氨基酸组成分析。结果显示,大豆蛋白酶解物经Sephadex G-25分离后得到了六个肽片段,其中平均链长为4,即含有2～6个氨基酸残基的大豆功能短肽SPP4,对羟自由基具有最佳的清除作用,清除率达到80．13%,氨基酸组成分析发现该组分中缬氨酸、亮氨酸、苯丙氨酸、赖氨酸含量较高。结果表明,含2～6个氨基酸残基的大豆生理活性短肽具有较好的抗氧化活性。