Effect of digestive enzymes on the bioactive properties of goat milk protein hydrolysates

The aim of this research was to study the influence of the gastrointestinal digestion on the bioactivity of goat milk protein hydrolysates prepared with subtilisin, trypsin and a combination of these two enzymes. All hydrolysates had excellent angiotensin converting enzyme (ACE) inhibitory activity, antioxidant activity and bile acid-binding capacity. Peptide profiles and bioactivities were mainly altered during the intestinal digestion, whereas the effect of the gastric digestion was negligible. The influence of the intestinal digestion varied depending on the hydrolysate and the bioactivity studied. In the case of ACE inhibitory activity, it exclusively decreased when peptides were produced with trypsin. In contrast, antioxidant activity and bile acid-binding capacity improved after the gastrointestinal digestion, regardless the enzymatic treatment conducted. Hydrolysis employing mixtures of subtilisin and trypsin is considered a good approach to produce peptides that maintain, or even enhance, their bioactivity after digestion.     

Identification and functional characterisation of bioactive peptides in rice bran albumin hydrolysates

The potential heath‐benefitting bioactivities of rice bran albumin hydrolysates were investigated. The antioxidant and α‐glucosidase‐ and angiotensin‐converting enzyme (ACE)‐inhibitory activities of the crude hydrolysates and their fractions were determined. The fractions with the highest bioactivities were analysed by liquid chromatography–tandem mass spectrometry/mass spectrometry to identify the active peptide sequences. Hydrolysates produced by commercial proteases Alcalase and Protamax exhibited the highest α‐glucosidase‐ and ACE‐inhibitory activities, resulting in 43.1 ± 2.1% and 54.4 ± 5.1% inhibition of the enzymes, respectively. Inhibitory activities against both enzymes were highest in the MW<3‐kDa fractions that were eluted under alkaline conditions. A number of peptide sequences were identified in the fractions, which contained several sequences with reported α‐glucosidase‐ or ACE‐inhibitory activities. This is the first time that such activities are reported for rice bran albumin hydrolysate, and it demonstrated that the hydrolysates may be developed into nutraceuticals useful in managing diabetics and hypertension.     

Antioxidant activities and functional properties of soy protein isolate hydrolysates obtained using microbial proteases

Soy protein isolate (SPI) hydrolysates were prepared using microbial proteases to produce peptides with antioxidant activity. The process parameters (substrate and enzyme concentrations), hydrolysis time, functional properties and the effects of ultrafiltration were further investigated. The results showed that the soy protein isolate exhibited a 7.0‐fold increase in antioxidant activity after hydrolysis. The hydrolysis parameters, defined by the experimental design, were a substrate concentration of 90 mg mL^?1 and the addition of 70.0 U of protease per mL of reaction. The maximum antioxidant activities were observed between 120 and 180 min of hydrolysis, where the degree of hydrolysis was approximately 20.0%. The hydrolysis increased solubility of the soy protein isolate; however, the hydrolysates exhibited a tendency to decrease in the interfacial activities and the heat stability. The SPI hydrolysates fractions obtained by ultrafiltration showed that the enzymatic hydrolysis resulted in samples with homogenous size and strong antioxidant activity.     

水解蛋白来源的抗氧化肽研究进展

采用水解技术制备抗氧化肽可大大拓宽其来源。本文介绍了影响水解技术制备抗氧化肽的因素，综述了国内外在这一领域的研究进展，并展望了此种方法制备抗氧化肽产业化的可能性。     

Understanding glycation kinetics of individual peptides in protein hydrolysates

Protein hydrolysates contain peptides with different lengths, type (α/ε-) and number of amino groups; these properties might influence the peptide glycation kinetics during the Maillard reaction. To identify the effects of peptide properties and hydrolysate composition on glycation kinetics, the glycation kinetics of individual peptides in hydrolysates was followed using quantitative peptide analysis. α-Lactalbumin was hydrolysed and glycated with d-glucose (0–8 h, 50 °C, dry heating with 65% humidity). The hydrolysates (degree of hydrolysis 2, 4, 6, and 8%) contained 25 unique peptides, ranging from 2 to 123 AAs with 0–12 lysine(s). The glycation rate constant (k_g) and the maximum average degree of glycation (DG_P_av,max) of peptides were independent of the hydrolysate composition. The maximum DG of α-NH₂ and ε-NH₂ groups was 12.8% and 60.0%, respectively. With this information, the DG_P_av,max of individual peptides [9–59% for peptides with 0–2 lysine(s)] could be predicted.     

不同蛋白酶水解棉籽蛋白制备抗氧化多肽的研究

利用6种蛋白酶对棉籽蛋白进行酶解,测定了各酶在水解过程中的水解度及其变化,对酶解产物的抗氧化活性进行了分析比较。研究表明,各蛋白酶在水解的前2h内,水解度迅速增加,2h之后水解曲线变得平缓。其中胃蛋白酶的水解能力最强,其4h水解产物水解度最大,为30.40%;胰蛋白酶的水解能力最差,最终水解产物的水解度为17.61%。中性蛋白酶水解产物的抗氧化活性较强,经测定其DPPH清除能力为54.95%,羟自由基清除能力为68.98%,超氧阴离子自由基清除能力为58.38%。     

Innovative applications of high-intensity ultrasound in the development of functional food ingredients: Production of protein hydrolysates and bioactive peptides

In recent years, the research on functional peptide generation for the development of functional foods has focused, among other issues, on the enhancement of enzymatic hydrolysis by means of high-intensity ultrasound (HIU) application. It has been suggested that the use of HIU in pretreatment and during the hydrolysis process can modify protein conformation by affecting hydrogen bonds and hydrophobic interactions, disrupting the quaternary and/or tertiary structure of proteins due to the effects of cavitation. Therefore, these structural modifications may expose more hydrolysis sites to be accessible by the enzyme, causing an increase in degree of hydrolysis and bioactivity. The main objective of this work was to review recent advances in food science and technology on the application of HIU for the enhancement of enzymatic hydrolysis of proteins of both animal and plant origin in order to generate novel protein hydrolysates and bioactive peptides which could be used as functional-food ingredients.     

菜籽蛋白活性肽研究

目前国内外研究表明,菜籽蛋白通过适当蛋白酶水解或经一定化学改性可得到大量具有生物活性和功能特性肽类,主要为抗氧化肽、降血压肽等;这些活性肽发现为今后充分利用菜籽蛋白资源提供新的途径。     

Antihypertensive activity of fish protein hydrolysates and its peptides

ABSTRACT

The rising interest to utilize nutritionally exorbitant fish proteins has instigated research activities in fish waste utilization. The development of newer technologies to utilize fish waste has fostered use of bioactive value-added products for specific health benefits. Enzymatically obtained Fish Protein Hydrolysate (FPH) is a rich source of biologically active peptides possessing anti-oxidant, anticancer, antimicrobial and anti-hypertensive activity. Isolating natural remedies to combat alarming negative consequences of synthetic drugs has been the new trend in current research promoting identification of antihypertensive peptides from FPH. In this review, we aim to culminate data available to produce antihypertensive peptides from FPH, its composition and potential to be used as a therapeutic agent. These purified peptides are known to be rich in arginine, valine and leucine. Reports reveal peptides with low molecular weight (<1 kDa) and shorter chain length (<20 amino acids) exhibited higher antihypertensive activity. As these peptides have proven Angiotensin Converting Enzyme – I inhibitory activity in vitro and in vivo, their potential to be used as antihypertensive drugs is outrageous. However, current focus on research in the field of molecular docking is necessary to have improved understanding of interaction of the peptides with the enzyme.     

Influence of the degree of hydrolysis on the bioactive properties of whey protein hydrolysates using Alcalase®

Whey protein concentrate was enzymatically hydrolysed at several time courses using the commercial preparation Alcalase^® 2.4 L, different hydrolysates were achieved, and the effect of degree of hydrolysis (DH) on both technological and biological properties was studied. Results have shown that solubility, antioxidant and ACE inhibition activities were increased as DH was also augmented from about 8 to 17%. RP‐HPLC studies also revealed a decrease in hydrophobicity when samples were hydrolysed in comparison with controls. When the enzyme hydrolytic action was augmented, it stimulated both the bioactivity of whey protein and relevant technological properties, allowing these hydrolysates to be employed as additives in the development of food formulations.     

乳源活性肽的细胞生长调节作用

乳源性细胞生长调节肽是生物活性肽领域重要的研究方向,它在抑制肿瘤细胞的生长、刺激免役细胞活性、诱导肠道细胞的凋亡发挥着重要作用。综述了乳蛋白活性肽对细胞生长调节功效的最新研究进展。     

酶解蛋白产物脱苦技术研究进展

酶水解蛋白产物具有极高营养价值及多种功能特性;但呈显苦味一直是困扰其在食品工业应用重要问题。该文就蛋白酶解产物苦味产生机理及苦味脱除技术进行综述。     

大豆源生物活性肽的研究进展

与具有相同氨基酸组成的大豆蛋白质相比,大豆蛋白酶解物中的大豆多肽具有许多独特的理化特性与生物学活性.目前,从大豆蛋白中已分离出多种纯化的大豆生物活性肽,如降血压肽、免疫调节肽、抗氧化肽等,对这些大豆活性肽的研究情况进行了综述,以期为大豆活性肽的研究开发提供基础资料.     

Dual function peptides from pepsin hydrolysates of whey protein isolate

The aim of this study was to investigate the effect of pepsin hydrolysates of whey protein isolate (WPI) on vascular relaxation and emulsifying capacity. WPI was subjected to pepsin hydrolysis for 5 h. The chromatographic profiles of the samples showed the formation of a wide variety of peptides. Addition of WPI hydrolysates in phenylephrine-contracted rat aortic rings induced a similar concentration-dependent relaxation in both endothelium-intact and endothelium-denuded preparations. In endothelium-denuded vessels the maximum relaxation induced by WPI fractions increased along the time, reaching over 70% after 3 h-hydrolysis on. In addition, the vascular relaxation was not associated with an inhibition of the angiotensin-converting enzyme or activation of K⁺ channels. Hydrolysed fractions were further evaluated for the emulsifying capacity (EC) and all tested fractions were able to keep an EC over 60%. These results reinforce the potential of WPI pepsin-hydrolysates as an option in the search for dual function peptides from whey proteins.     

Preparation and analysis of hydrolysates from whey protein concentrate using the proteases from Bacillus licheniformis and Aspergillus oryzae

The action of proteases from Bacillus licheniformis and Aspergillus oryzae was studied with the aim of preparing hydrolysates from whey protein concentrate with nutritionally appropriate peptide profile. Various enzyme/substrate ratios were used, and the peptides were fractionated by size‐exclusion HPLC followed by their quantification using the rapid method of correct fraction area (CFA). The protease from B. licheniformis (E:S of 8:100) produced the best peptide with a much lower amount of large peptides (44.61%), greater of di‐ and tripeptides (8.79%) and higher sum of the di‐ and tripeptides with free amino acids (9.99%) than the other hydrolysates. The advantage of using a lower E:S ratio to obtain a nutritionally adequate peptide profile was observed for the protease of A. oryzae when it passed from 3:100 to 2:100.     

番茄籽蛋白酶解产物的功能特性研究

以番茄籽蛋白为原料,研究不同蛋白酶对番茄籽蛋白水解物功能性质的影响并与原蛋白的功能性质进行比较。结果表明:不同蛋白酶水解番茄籽蛋白的规律相似,水解度随时间增加而提高,但在相同时间内,不同水解物的水解度不同,说明不同的蛋白酶对番茄籽蛋白的水解效率不同,其中碱性蛋白酶水解效率最高14.48%;木瓜蛋白酶可较高效地回收番茄籽蛋白,蛋白回收率高达81.12%;番茄籽分离蛋白经酶解改性后其表面疏水性降低,但溶解性和乳化性均有不同程度的提高,其中胰蛋白酶水解物的溶解性最好;木瓜蛋白酶水解物的乳化性能最优。因此,酶法水解可改善番茄籽蛋白的功能性质,且不同蛋白酶对蛋白的功能性质的影响不同,可根据实际需要选择蛋白酶。     

The presence of bioactive peptides in hydrolysates prepared from processing waste of sardine (Sardina pilchardus)

A proteolytic enzyme, Alcalase®, was used to produce partly digested proteins from cooked wastes of sardine (Sardina pilchardus). The presence of biologically active molecules was investigated in these hydrolysates prepared under various conditions of time and enzyme/substrate ratio. By means of radioimmunoassays and mitogenic and radioreceptor assays, the presence of molecules related to secretagogue peptides, growth factors and calcitonin gene‐related peptide (CGRP) respectively was detected in the hydrolysates. Exclusion chromatography permits the conclusion that the biological activity of the different fractions is related to their size. © 2001 Society of Chemical Industry     

番茄籽蛋白酶解产物的功能特性研究

以番茄籽蛋白为原料,研究不同蛋白酶对番茄籽蛋白水解物功能性质的影响并与原蛋白的功能性质进行比较。结果表明:不同蛋白酶水解番茄籽蛋白的规律相似,水解度随时间增加而提高,但在相同时间内,不同水解物的水解度不同,说明不同的蛋白酶对番茄籽蛋白的水解效率不同,其中碱性蛋白酶水解效率最高14.48%;木瓜蛋白酶可较高效地回收番茄籽蛋白,蛋白回收率高达81.12%;番茄籽分离蛋白经酶解改性后其表面疏水性降低,但溶解性和乳化性均有不同程度的提高,其中胰蛋白酶水解物的溶解性最好;木瓜蛋白酶水解物的乳化性能最优。因此,酶法水解可改善番茄籽蛋白的功能性质,且不同蛋白酶对蛋白的功能性质的影响不同,可根据实际需要选择蛋白酶。      

Three novel angiotensin I-converting enzyme (ACE) inhibitory peptides from cuttlefish (Sepia officinalis) using digestive proteases

The angiotensin I-converting enzyme (ACE) inhibitory activities of protein hydrolysates prepared from muscle of cuttlefish (Sepia officinalis) by treatment with various digestive proteases were investigated. The most active hydrolysate was obtained with the crude protease extract from the hepatopancreas of cuttlefish (64.47 ± 1.0% at 2 mg of dry weight/ml) with a degree of hydrolysis of 8%. By gel filtration on Sephadex G-25 and RP-HPLC on C18 column, three novel peptides with high ACE-inhibitory activity were purified and their molecular masses and amino acid sequences were determined. The three peptides Val-Tyr-Ala-Pro, Val-Ile-Ile-Phe and Met-Ala-Trp with IC₅₀ values of 6.1, 8.7 and 16.32 μM, respectively, were novel ACE-inhibitory peptides. Lineweaver–Burk plots suggest that the three purified peptides act as non-competitive inhibitors against ACE. These results suggest that some peptides from cuttlefish could be a beneficial ingredient for nutraceuticals against hypertension.