Functional Properties of Fish Protein Hydrolysate from Herring (Clupea harengus)

Raw herring (Clupea harengus), a waste product from the roe industry, was hydrolyzed, using an endopeptidase preparation from Bacillus licheniformis. Aliquots were taken at 0, 5, 10, 20, 30, 45, and 60 min hydrolysis to measure the degree of hydrolysis. The functional properties tested were emulsifying activity index (EAI), foamability, and foam stability of the hydrolysate. At 36% hydrolysis, the herring hydrolysate presented good emulsifying stability (> 120 minutes) and an adequate foam expansion (142%), as compared to the soluble fraction from the unhydrolyzed control herring. The lipid content decreased considerably to 0.77% for the fish protein hydrolysate, while its protein content increased to 77%. The amino acid composition remained similar to that of the control.     

Cadmium-binding protein components of flaxseed: Influence of cultivar and location

The distribution of cadmium-(Cd)-binding components from five flaxseed cultivars grown at three locations in southern Manitoba was investigated to examine genotypic and environmental effects. Three protein fractions with different electrostatic properties, eluting at 0.10, 0.25 and 0.50 M NaCl by ion-exchange chromatography DEAE-Sephacel, represented 12%, 66% and 7% of the bound (extracted) protein, respectively, while 15% of the protein remained unbound. Cadmium and other divalent metal (zinc, copper and calcium) contents of protein fractions were strongly influenced by location. Cultivar differences in protein and cadmium contents of the protein fractions were highly significant. Cadmium and zinc accumulated similarly in the 0.10 and 0.25 M protein fractions at 51% and 40–43%, respectively. Transfer of copper occurred prominently in the 0.50 M fraction while most of the calcium (55%) remained unbound. The distribution of cadmium, zinc, copper and calcium in fractions of flaxseed proteins was strongly influenced by cultivar and location, indicating differences in their accumulation, migration and transfer.     

Antiproliferative activity of peptides prepared from enzymatic hydrolysates of tuna dark muscle on human breast cancer cell line MCF-7

To produce and identify antiproliferative peptides, two commercial enzymes, papain (PA) and Protease XXIII (PR) were used to hydrolyse tuna dark muscle byproduct, and the protein hydrolysates were purified, before being evaluated for antiproliferative activities against human breast cancer cell line MCF-7. The results showed that the peptide fractions with the molecular weight ranging from 390 to 1400 Da possessed the greatest antiproliferative activity. The amino acid sequences of the two antiproliferative peptides isolated from PA and PR hydrolysates were Leu-Pro-His-Val-Leu-Thr-Pro-Glu-Ala-Gly-Ala-Thr (1206 Da) and Pro-Thr-Ala-Glu-Gly-Gly-Val-Tyr-Met-Val-Thr (1124 Da), whilst they show the dose-dependent inhibition effect of the MCF-7 cells with IC₅₀ values of 8.1 and 8.8 μM, respectively. We thus conclude that antiproliferative hydrolysates from tuna dark muscle byproduct may be useful ingredients in food and nutraceutical applications.     

Angiotensin-I-converting enzyme inhibitory activity and bitterness of enzymatically-produced hydrolysates of shrimp (Pandalopsis dispar) processing byproducts investigated by Taguchi design

The importance of water-to-substrate ratio, protease type, percent enzyme and incubation time on hydrolysates produced from shrimp processing byproducts was investigated using Taguchi’s L₁₆ (4⁵) experimental design. Protease type significantly (p < 0.05) influenced soluble yield, degree of hydrolysis (DH), angiotensin-I-converting enzyme (ACE) inhibitory activity and bitterness of hydrolysates, while percent enzyme only affected the DH. Hydrolysates produced by Alcalase and Protamex possessed strong ACE inhibitory activity (IC₅₀ = 100–200 μg/ml and 70 μg/ml, respectively), accompanied by high yield, high DH and strong bitterness. Furthermore, ACE inhibition was positively correlated (r² = 0.87) with bitterness of the hydrolysates. Fractionation by size-exclusion chromatography revealed that the bitter substances, which also showed strong ACE inhibition, were <3 kDa in size and contained many hydrophobic residues, including Tyr, Phe, Leu, Ile, Val and Lys. Despite the bitterness, these hydrolysates may have potential health benefits, arising from their potent ACE inhibitory activity.     

Angiotensin I-Converting Enzyme Inhibitory Activity of Soy Protein Digests in a Dynamic Model System Simulating the Upper Gastrointestinal Tract

ABSTRACT: The objective of this study was to investigate whether peptides with inhibitory activity against angiotensin I-converting enzyme (ACE) would be produced by digestion of isolated soy protein (ISP) in a dynamic model system simulating gastrointestinal conditions. Using the model system, 5% ISP solution was pumped into the stomach reactor containing pepsin and HCl. The peptic digest was continuously pumped into the duodenum reactor containing pancreatin and Oxgall bile. The effects of blanching (100°C, 10 min) followed by pasteurization (75°C, 15 s) or sterilization (121°C, 20 min) of ISP before digestion on the inhibitory activity were also investigated. During the first 30 min of digestion, significantly higher ( P < 0.05) ACE-inhibitory activity was generated from unheated ISP after sequential digestion in both reactors compared with after peptic digestion only in the stomach reactor. However, at 90 min, subsequent digestion by pancreatin of unheated and blanched-sterilized ISP decreased ACE-inhibitory activity compared with peptic digestion alone. The IC₅₀ values at the end of 90 min digestion in both reactors were 0.38 ±0.01, 0.37 ± 0.02, and 0.44 ± 0.02 mg/mL for unheated, blanched-pasteurized, and blanched-sterilized ISP, respectively. The results suggest the potential production of peptides with ACE-inhibitory activity upon physiological digestion of soy protein, including products that have been subjected to heat processing. Although clinical trials would be required to provide final evidence of efficacy of the soy peptides, the present findings support the application of soy protein as an ingredient for functional foods.     

The role of molecular size in antioxidant activity of peptide fractions from Pacific hake (Merluccius productus) hydrolysates

The antioxidative properties of Pacific hake hydrolysates and their peptidic fractions varying in molecular size were assessed. Hydrolysates produced by different proteases (Alcalase, bromelain, Flavourzyme, Protamex, Protease A“Amano”2, Protease N“Amano”K, Protin SD NY10, Umamizyme-K, Validase BNP-L, Validase FPexo) generally possessed good metal ion chelating (33–73% at 3 mg/ml), DPPH radical scavenging (18–30% at 1 mg/ml), ferric ion reducing power (abs_700nm 0.36–0.86 at 3 mg/ml) and ABTS radical scavenging (47–85% at 0.067 mg/ml) activity, as well as a good capability to suppress lipid peroxidation in a linoleic acid model system. Peptide size (<1.4 kDa) was important for ABTS radical scavenging activity, whereas specific peptide composition (which depended on the particular protease used) was the governing factor for effective lipid peroxidation. Validase BNP-L was the most promising enzyme for producing Pacific hake hydrolysates with good antioxidative activity in various assays and similar effectiveness as the synthetic antioxidant BHT to inhibit lipid peroxidation.     

Pacific Hake (Merluccius Productus) Hydrolysates as Cryoprotective Agents in Frozen Pacific Cod Fillet Mince

ABSTRACT:  Fish protein hydrolysates produced by proteolysis of Pacific hake ( Merluccius productus ) with Alcalase^® (FPH-A) or Flavourzyme^® (FPH-F) were investigated as a potential alternative to the 1 : 1 blend of sucrose–sorbitol (SuSo) commonly used for cryoprotection of frozen fish mince. The physicochemical properties of cod mince samples in the absence (control) or presence of 8% FPH-A, FPH-F, or SuSo were evaluated before and after 6 freeze–thaw cycles, with differences noted at the 5% significance level. Freeze–thawing of control sample increased expressible moisture (from 22% to 33%) and cook loss (from 3% to 16%). These poor water retention properties were improved in samples containing FPH or SuSo. Differential scanning calorimetry showed higher proportion of unfrozen water in freeze–thawed samples containing FPH-F or FPH-A (0.36 g/g) compared to SuSo (0.33 g/g) and control (0.24 g/g) samples. Textural analysis of cooked mince from unfrozen samples indicated greater hardness for FPH than SuSo and control samples, while freeze–thawing resulted in decreased hardness for FPH and SuSo samples. Content and surface hydrophobicity of extractable natural actomyosin (NAM) were maintained after freeze–thawing of samples containing FPH-F or SuSo, compared to 50% decrease in extractable NAM and a significant increase in surface hydrophobicity for the control. The presence of oligopeptides in both hydrolysates and the high levels of free amino acids including Asp, Glu, Arg, and Lys in FPH-F might be responsible for their cryoprotective action. This study provides strong evidence to support development of FPH as a new generation cryoprotectant to maintain quality of frozen fish.     

Investigation of the Putative Associations Between Dairy Consumption and Incidence of Type 1 and Type 2 Diabetes

A growing body of evidence suggests a possible relationship between the consumption of dairy products and the incidence of diabetes. A positive correlation between the early introduction of dairy in infancy and the incidence of type 1 diabetes (T1D) in genetically predisposed infants has been suggested by studies on rodents and humans. However, the lines of evidence supporting this association, including epidemiological studies and the observation of antibodies to bovine serum albumin, β-casein and bovine insulin in the serum of patients with T1D, are not without controversy. On the other hand, an inverse relationship between the consumption of dairy foods and the development of metabolic syndrome and/or type 2 diabetes (T2D) has been implied by epidemiological studies. Several dairy components, especially milk proteins, are believed to play a role in the beneficial effect of dairy consumption on glucose regulation by modulation of incretin hormones. Other dietary factors have also been associated with the incidence of T1D and T2D, indicating that dairy foods might be only one among many dietary agents possibly implicated in the development of diabetes. The present paper critically reviews the evidence and plausible mechanisms for the putative associations between dairy food consumption and incidence of T1D and T2D.     

The prebiotic arabinogalactan of Anoectochilus formosanus prevents ovariectomy-induced osteoporosis in mice

Anoectochilus formosanus (Orchidaceae) has previously been shown to exhibit anti-osteoporosis and prebiotic properties. In this study, these bioactivities were verified and associated with an isolated type II arabinogalactan (AGAF) in ovariectomized (OVX) mice model. Female ICR mice were OVX and administrated AGAF (5 and 15 mg/kg) or inulin (400 mg/kg) orally for 3 weeks. Streptomycin was used for blocking the bioactivities of AGAF. In results, AGAF increased the level of fecal bifidobacteria, cecal soluble Ca and short chain fatty acids. Comparing to OVX control group, AGAF improved bone mineral content, trabecular bone volume, and the number of trabecular significantly. In RT-PCR analysis, AGAF reduced the expression of tartrate-resistant acid phosphatase, cathepsin K, and osteocalcin. Streptomycin inhibited both anti-osteoporosis and prebiotic effects of AGAF. In vitro experiments revealed butyrate, not AGAF could activate osteoblasts and inhibit osteoclasts differentiation. Thus, this study demonstrated that AGAF prevents bone loss in OVX mice through prebiotic effects in vivo and in vitro.     

Interactions between aroma compounds and β-lactoglobulin in the heat-induced molten globule state

The present study aims to elucidate the binding of small hydrophobic ligands onto the molten globule state of β-lactoglobulin (BLG). The conversion of the native BLG into a molten globule state was induced by heat treatment at acidic pH. The molten globule state was evidenced by far and near-UV circular dichroism spectra. β-Ionone and guaiacol exhibited a higher binding ability to BLG in the heat-induced molten globule state compared to unheated BLG, as assessed by protein surface hydrophobicity measurements, using 6-propionyl-2-(dimethylamino)naphthalene (PRODAN) fluorescent probe. The binding sites of the two aroma compounds were determined by 2D nuclear magnetic resonance (NMR) spectroscopy. The less tightly packed structure of the molten globule favoured ligand binding, in particular within the central cavity. The greater flexibility of the calyx entrance, and the conformational change of loop EF induced an easier access of the central cavity after the thermal treatment.