Enzymatic production of bioactive peptides from milk and whey proteins (original) (raw)
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Enzymatic hydrolysis of whey protein concentrate was used to prepare hydrolysates with appropriate peptide profiles for use in the development of dietary supplements. Effects of the type of enzyme (pancreatin and proteases from Bacillus licheniformis, Aspergillus oryzae and Aspergillus sojae) and enzyme:substrate ratio (0.5:100, 1:100, 2:100, 3:100, 4:100 and 8:100) were evaluated. Twenty-four hydrolysates were prepared, and their peptide profiles were characterized by chain length using size exclusion liquid chromatography fractionation, followed by a rapid Correct Fraction Area method for quantifying the peptide and amino acid contents. The use of a protease from Bacillus licheniformis (E:S 8:100) and a pancreatin (E:S 2:100, 3:100, 4:100 and 8:100) yielded preparations of hydrolysates with the highest sums of di-and tripeptide and free amino acid contents (9.99 and 11.28 % for the protease from Bacillus licheniformis and pancreatin, respectively) as well as the lowest amount of large peptides (44.61 and 44.13 %, respectively).
Journal of Food Science and Technology, 2013
Enzymatic hydrolysis of whey protein concentrate was used to prepare hydrolysates with appropriate peptide profiles for use in the development of dietary supplements. Effects of the type of enzyme (pancreatin and proteases from Bacillus licheniformis, Aspergillus oryzae and Aspergillus sojae) and enzyme:substrate ratio (0.5:100, 1:100, 2:100, 3:100, 4:100 and 8:100) were evaluated. Twenty-four hydrolysates were prepared, and their peptide profiles were characterized by chain length using size exclusion liquid chromatography fractionation, followed by a rapid Correct Fraction Area method for quantifying the peptide and amino acid contents. The use of a protease from Bacillus licheniformis (E:S 8:100) and a pancreatin (E:S 2:100, 3:100, 4:100 and 8:100) yielded preparations of hydrolysates with the highest sums of di-and tripeptide and free amino acid contents (9.99 and 11.28 % for the protease from Bacillus licheniformis and pancreatin, respectively) as well as the lowest amount of large peptides (44.61 and 44.13 %, respectively).
International Journal of Food Science & Technology, 2017
Novel bacterial (HT) and fungal (FPII) food-grade protease preparations were evaluated for their ability to hydrolyse sheep cheese whey (SCW) and the generation of bioactive peptides. Both protease preparations hydrolysed the whey proteins to small peptides over 24-h hydrolysis time, but the time course hydrolysis profiles were different as evaluated by SDS-PAGE. The HT whey hydrolysate had considerably higher antioxidant and angiotensin-I converting enzyme (ACE)-inhibitor activity than the FPII hydrolysate. Neither hydrolysate was cytotoxic towards Vero cells. OFFGEL electrophoresis of the small peptide pool fraction (<15 amino acids) of each hydrolysate indicated differences in the pI distribution of the bioactive peptides. This likely reflects the diverse hydrolytic specificity of the proteases. Although the antioxidant activity of both hydrolysates was not significantly affected by simulated gastrointestinal digestion, the loss of ACE-inhibitor activity was greater with the FPII hydrolysate.
A comparison of dual-functional whey hydrolysates by the use of commercial proteases
Food Science and Technology
Whey is well-known for the functional and bioactive properties of its proteins and peptides, which are of great interest to food and nutraceutical industries. The aim of this study was to investigate the effect of four different commercial proteases (Novo ProD (NPD), Alcalase (ALC), Pancreas Trypsin (TRY), and Flavourzyme (FLA)) in the generation of hydrolysates with emulsifying and antioxidant activities. Hydrolysis processes were carried out for 5 h, reaching maximum degrees of 18.5, 15.5, 9.2, and 8.7% for NPD, ALC, TRY, and FLA, respectively. All tested enzymes generated very diverse, but conservative peptide profiles when comparing the treatments along the time, with the main enzymatic actions up to 120 min-reaction. An increase in the in vitro antioxidant activity was found for all treatments, achieving 46%, 40%, 40% and 22% for ALC, TRY, NDP and FLA, respectively. TRY hydrolysate maintained or slightly increased its emulsifying capacity along the time, however, a decrease in emulsifying capacity was found for ALC and NPD hydrolysates when compared to the non-hydrolyzed whey protein concentrate (WPC). All tested enzymes generated hydrolysates with enhanced antioxidant and/or emulsifying activities, which may be used as food ingredients and the choice of the enzyme will depend on the need.
LWT - Food Science and Technology, 2015
A crude extract containing serine peptidases, was prepared from latex of Maclura pomifera fruits. Peptidases were isolated by precipitation with one volume of ethanol with a yield of 5.4 ± 0.4 Ucas per milligram of protein. This extract was used for hydrolysis of bovine whey proteins at 45 C and pH 6.5. Proteolytic activity was 99% inactivated after 5 min of heat treatment (100 C). Major whey proteins degradation profile was analysed by tricine SDS-PAGE. After 180 min of hydrolysis alpha-lactalbumin (a-LA) and beta-lactoglobulin (b-LG) were almost completely degraded. Hydrolysis degree was 31.3 ± 1.7% at 180 min of reaction and the peptides produced that were smaller than 3 kDa were analysed by reversedphase high-performance liquid chromatography (RP-HPLC). Angiotensin-converting enzyme (ACE) inhibitory activity and antioxidant capacity were detected in the hydrolysates and IC 50 values for 180 min of hydrolysis were 0.53 ± 0.02 and 4.44 ± 0.44 mg/ml, respectively. One peptide sequence deduced from peptide masses in the 180 min filtered hydrolysate, coincided with an ACE-inhibitory peptide reported by other author. The results support the conclusion that, by the presence of ACE-inhibitory and antioxidant peptides, it would be possible to use these whey protein hydrolysates for functional food manufacturing.
Milk peptides; effect on the enzymatic hydrolysis of sodium caseinate
Agricultural and Food Science, 1993
Sodium caseinate (NaCN) was hydrolyzed using Rhozyme 41 (Rh41), Neutrase (Neu) and plasmin (PL) to obtain peptide preparations termed; Na-Cas-P-Rh41, Na-Cas-P-Neu, and Na-Cas-P-Plasmin, respectively. Indigenous whey peptides (IWP) were obtained from fresh sweet whey, at different levels of ufconcentration, by a precipitation method described earlier. These peptide fractions were then used to observe their effect on the activity of some proteases. All peptide preparations depressed enzyme activity. Na-Cas-P-Rh41 was the most powerful inhibitor of enzyme activity and decreased the activity of trypsin, Rh41, Neu. and PL by 15, 32, 50, and 14%, respectively. IWP markedly depressed activity of Rh41. The degree of uf-concentration of whey from which IWP was obtained was directly related to degree of inhibition.
Journal of Agricultural and Food Chemistry, 2020
There is a wide variety of peptides released from food proteins which are able to exert a relevant benefit for human health like angiotensin-converting enzyme (ACE) inhibition, antioxidant, anti-inflammatory, hypoglucemic or antithrombotic activity, among others. This manuscript is reviewing the recent advances on enzymatic mechanisms for the hydrolysis of food proteins, including the types of enzymes and mechanisms of action involved, the strategies followed for the isolation and identification of bioactive peptides through advanced proteomic tools, the assessment of bioactivity and its beneficial effects. Specific applications in fermented and/or ripened foods where a significant number of bioactive peptides have been reported with relevant in vivo physiological effects on laboratory rats and humans, as well as the hydrolysis of food proteins for the production of bioactive peptides are also reviewed.
International Journal of Food Science & Technology, 2013
The action of various proteases was tested for preparing whey protein concentrate (WPC) hydrolysates with high degree of hydrolysis (DH), appropriate peptide profiles and reduced phenylalanine (Phe) content. The peptide profile analysis included the fractionation of hydrolysates by size-exclusion HPLC. The rapid correct fraction area method was used to quantify the components of the chromatographic fractions. Activated carbon (AC) was used to remove Phe, and its efficiency was evaluated by measuring the amount of Phe by second-derivative spectrophotometry. The results showed that the DH of WPC hydrolysates increased and that the protease from Aspergillus oryzae yielded the highest DH value. This protease also produced the best peptide profile, that is, the highest di-and tripeptide content (16.14%), the highest amounts of free amino acids (18.43%) and the lowest amount of large peptides (18.76%). The proteases from both A. oryzae and Bacillus subtilis produced the highest Phe removals (79.0 and 77.8%, respectively). Analysis of whey protein hydrolysates M. P. C. Silvestre et al. 589 Analysis of whey protein hydrolysates M. P. C. Silvestre et al. Analysis of whey protein hydrolysates M. P. C. Silvestre et al. Analysis of whey protein hydrolysates M. P. C. Silvestre et al. Analysis of whey protein hydrolysates M. P. C. Silvestre et al. Analysis of whey protein hydrolysates M. P. C. Silvestre et al. Analysis of whey protein hydrolysates M. P. C. Silvestre et al.
Enzymatic Hydrolysis of Whey Protein Concentrates: Peptide HPLC Profiles
Journal of Liquid Chromatography & Related Technologies, 2004
Hydrolysis of whey protein concentrates (WPCs) at different temperatures and pHs, using three enzymes: pepsin, trypsin, and Alcalase w , was monitored during more than 5 hr by reversed phase HPLC/UV, using a column containing a polystyrene-divinylbenzene copolymer-based packing, and an elution gradient from 8% to 80% acetonitrile containing 0.1% TFA. Peptides were separated according to their polarity and size, and degradation of a-lactalbumin (a-la) and b-lactoglobulin (b-lg) was evaluated. The three proteolytic enzymes (pepsin, trypsin, and Alcalase w ) employed for hydrolysis of WPCs led to different kinetics of degradation of b-lg. a-la degradation after 15 min was almost complete for the three enzymes. The hydrolysis catalysed by each enzyme resulted in different peptide profiles by HPLC/UV. Hydrolysates produced by pepsin (HP) were resolved into three main fractions of high retention times, while tripsin hydrolysates (HT) were resolved into nine major peaks and Alcalase w hydrolysates (HA) were resolved into 12 major peaks, presenting a wide range of polarities and sizes. Although, with different b-lg hydrolysis extension, chromatographic profiles of the degradation and formation of peptides can be used as a finger print of the type of enzyme used, because peptide profile is not affected either by temperature or pH.
Food chemistry, 2017
An aspartic protease from Salpichroa origanifolia fruits was successfully immobilized onto an activated support of glutaraldehyde agarose. The immobilized enzyme presented higher thermal stability than the free enzyme from 40°C to 50°C and high reusability, retaining 54% of the initial activity after ten cycles of the process. Whey protein concentrates (WPC) were hydrolyzed with both free and immobilized enzyme, reaching a similar degree of hydrolysis of approximately 6-8% after 20h. In addition, the immobilized derivate hydrolyzed α-lactalbumin protein with a higher affinity than β-lactoglobulin. The hydrolysate was ultra-filtrated, and the fractions were evaluated for antioxidant activities with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity method. The fraction containing peptides with a molecular mass below 3kDa demonstrated a strong radical quenching effect (IC50: 0.48mg/ml). These results suggest that hydrolyzed WPC could be considered as a promising sour...