Modification of Soybean Protein Hydrolysates by Alcalase-Catalyzed Plastein Reaction and the ACE-Inhibitory Activity of the Modified Product In Vitro

Soybean protein hydrolysates were prepared by hydrolyzing soybean protein isolates with a protease alcalase to a degree of hydrolysis of 16.6%, and then modified by alcalase-catalyzed plastein reaction to reveal the impact of plastein reaction on the ACE-inhibitory activity of the modified product in vitro. The suitable conditions of plastein reaction of soybean protein hydrolysates were selected based on the results of response surface methodology with the decreased amount of the free amino groups of the modified product as response. When reaction temperature was fixed at 30°C, the selected conditions were as follows: concentration of soybean protein hydrolysates of 45% (w/w), addition level of alcalase of 275 U/g peptides, and reaction time of 3 to 4 h. Soybean protein hydrolysates and eight modified products were evaluated for their ACE-inhibitory activities in vitro. The assay results highlighted that plastein reaction improved the ACE-inhibitory activity of the modified product. The IC₅₀ of the modified products ranged from 0.64 to 1.11 mg/mL, while that of soybean protein hydrolysates was 1.45 mg/mL. The decreased amount of the free amino groups of the modified product showed influence on the ACE-inhibitory activity in vitro. Analysis results from size exclusion chromatography confirmed that some plasteins with higher molecular weights were formed in the modified product. Our results showed that alcalase-catalyzed plastein reaction could be applied as a potential approach to enhance the ACE-inhibitory activity of soybean protein hydrolysates in vitro.     

Antioxidant activity of several marine skin gelatins

Gelatins obtained from the skins of tuna and halibut, and from the tunics of jumbo flying squid were hydrolysed by Alcalase to produce antioxidant peptides. Hydrolysis yielded an increase in the antioxidant capacity of gelatins of around two-fold when measured by the Fe reducing capacity (FRAP) method and even more when measured by the ABTS radical scavenging method. When both squid and tuna gelatins were hydrolysed with different enzymes (collagenase, trypsin, pepsin), Alcalase and pepsin gave the hydrolysates with the highest and lowest ABTS radical scavenging ability, respectively. FRAP assay showed that the squid hydrolysates prepared using Alcalase were the most effective in reducing ferric ions, whereas trypsin gave rise to the tuna hydrolysates with the highest iron reducing ability. When the amino acid composition of the gelatins was related to the antioxidant properties, ABTS radical scavenging was observed to be negatively correlated to the total content of hydrophobic amino acids and imino acids in all the samples, while Fe reducing power (FRAP) was strongly correlated with Hyl and degree of hydroxylation.     

Application of ultrafiltration to the preparation of defined hydrolysates of bovine haemoglobin

Many uses of protein hydrolysates have been developed and applied to areas such as nutritional therapy, culture media, and the isolation of biologically active peptides. All these applications need carefully controlled and characterized hydrolysates. In order to produce such a type of hydrolysate, it is possible to use haemoglobin which is a very well defined and constant protein source. Enzymic hydrolysis of haemoglobin by pepsin was carried out at pilot-plant scale in an ultrafiltration reactor with mineral membranes. The object was to obtain a reproducible, decolorized, salt-free enzymic hydrolysate. Two types of membranes were tested having 10000 dalton (M5 type) and 20000 dalton (M4 type) cut-offs. Little significant difference was observed in the final products when both types of membranes were used. Reproducibility of hydrolysates was verified by amino acid analysis and gel filtration chromatography. The haemoglobin hydrolysates produced contained more than 90% protein and are especially suitable for fine applications.     

Oligosaccharides prepared by acid hydrolysis of polysaccharides from pumpkin (Cucurbita moschata) pulp and their prebiotic activities

As a rich source of polysaccharides, pumpkin pulp was used for the production of oligosaccharides. To investigate the prebiotic potential of the oligosaccharides, prepared by acid hydrolysis of polysaccharides from pumpkin pulp, the effect of the oligosaccharides on lactobacilli growth was studied. The oligosaccharides were composed of galactose (99.03%) and glucose (0.97%). The molecular weight distribution of the oligosaccharides showed a larger part of low molecular weight fraction in a range from 5000 to 324 Da than the polysaccharides, which gave the oligosaccharides higher solubility. The oligosaccharides showed good resistance to hydrolysis by artificial human gastric juice and α‐amylase, compared to the reference prebiotic inulin, and considerable capability to stimulate the growth of lactobacilli. In vitro studies showed that the number of lactobacilli was significantly greater than those in control medium when they were cultured in the medium supplemented with the oligosaccharides (P < 0.05), and there was no significant difference at the oligosaccharides group and the inulin group (P < 0.05).     

Preparation of wheat gluten hydrolysates with high opioid activity

The opioid activity of wheat gluten hydrolysates (WGHs) prepared with several proteases was investigated in vitro by the contraction of guinea pig ileum (GPI), and WGHs with high opioid activity were further obtained through desalting and ultrafiltration (UF). The opioid activity varied with different wheat gluten hydrolysates. AWGH (WGH prepared with Alcalase), PPWGH (WGH prepared with Pepsin followed by Pancreatin) and PWGH (WGH prepared with pepsin) had relatively stronger inhibitory effects on the contraction of GPI with IC₅₀ values of 1.21 ± 0.25, 1.29 ± 0.38 and 1.57 ± 0.21 mg protein/ml, respectively. Subsequently, AWGH and PPWGH were desalted using cation exchange resin and anion exchange resin, with high nitrogen recovery (88.51 and 89.28%, respectively) and high desalting ratio (84.67 and 85.20%). The resultant hydrolysates were further fractionated by UF performed with a 3 kDa molecular weight cut-off membrane. Compared with AWGH and PPWGH, the 3 kDa-permeates have higher opioid activities with high protein content above 90% and low ash content about 1.5%. The molecular weight distributions of the two 3 kDa-permeates were concentrated below 2,000 Da.     

Application of a Technofunctional Caseinate Hydrolysate to Replace Surfactants in Ice Cream

There is a demand for a clean label of ice cream in the food industry. To meet this requirement, alternative surfactants and hydrocolloids without an additive number have to be found. Here, a caseinate hydrolysate with improved interfacial properties was investigated as such an additive. Replacing only the surfactant resulted in ice cream characteristics, comparable to using a commercial emulsifier (INS 472b). Thus, a clean labeling of the surfactant using this hydrolysate is possible. By contrast, replacing the hydrocolloid additionally led to an unfavorable ice cream texture and stability.     

Effect of angiotensin I converting enzyme inhibitory peptide purified from skate skin hydrolysate

Our objective was to evaluate the angiotensin I converting enzyme (ACE) inhibitory activity of skate skin protein hydrolysates and its corresponding fractions. The skate skin hydrolysates were obtained by enzymatic hydrolysis using alcalase, α-chymotrypsin, neutrase, pepsin, papain, and trypsin. Amongst the six hydrolysates, the α-chymotrypsin hydrolysate had the highest ACE inhibitory activity compared to other hydrolysates. The amino acid sequences of the purified peptides were identified to be Pro–Gly–Pro–Leu–Gly–Leu–Thr–Gly–Pro (975.38 Da), and Gln–Leu–Gly–Phe–Leu–Gly–Pro–Arg (874.45 Da). The purified peptides from skate skin had an IC₅₀ value of 95 μM and 148 μM, respectively, and the Lineweaver–Burk plots suggest that they act as a non-competitive inhibitor against ACE. Our study suggested that novel ACE inhibitory peptides derived from skate skin protein may be beneficial as anti-hypertension compounds in functional foods.     

Preparation,characterisation and selected functional properties of hydrolysed sodium caseinate–maltodextrin conjugate

Sodium caseinate was hydrolysed to a limited, moderate or extensive degree. The hydrolysates were conjugated with maltodextrin by a Maillard‐type reaction by dry‐heat treatment at 60 °C and 79% relative humidity for 2 or 4 days. Conjugates were characterised by SDS–PAGE and gel permeation chromatography. In comparison with the hydrolysates themselves, the conjugated hydrolysates had improved solubility, particularly around the isoelectric pH of the protein. The emulsifying properties of these conjugates were assessed in oil‐in‐water (o/w) emulsions; on emulsion formation, each conjugate‐stabilised emulsion had lower mean fat globule size than the corresponding hydrolysate‐stabilised emulsion. After storage for 7 days under accelerated shelf life testing conditions, the limited and moderate hydrolysate conjugate–stabilised emulsions had improved storage stability compared with hydrolysate‐stabilised emulsions; however, further research is required to optimise the hydrolysate fraction prior to conjugation for the production of novel low molecular weight emulsifiers.     

Identification and characterization of multifunctional cationic peptides from enzymatic hydrolysates of soybean proteins

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Biologically active peptides obtained by enzymatic hydrolysis of Adzuki bean seeds

This study investigated the antioxidant and antihypertensive activities of peptides obtained from protein fractions of Adzuki bean seeds. Peptides were obtained by the use of hydrolytic enzymes in vitro under gastrointestinal conditions. A determination was made of the activity of the peptide inhibitors of the angiotensin I converting enzyme (ACE), and the antiradical and ion chelating activity of peptides from different protein fractions. The highest peptide levels after the absorption process (<7 kDa) were noted in the albumin fraction (50.69 μg/ml). Furthermore, it was found that peptides from the prolamin fraction were characterised by the highest antiradical activity and ACE inhibitory activity (IC₅₀ = 0.17 mg/ml). Peptides obtained from the globulin fraction showed the highest ability to chelate iron ions, and peptides from the glutelin fraction were characterised as being the most effective in the chelation of copper ions.