Impact of enzyme preparation and degree of hydrolysis on peptide profile and nitrogen solubility of sodium caseinate hydrolysates

Protein hydrolysates show great promise as bioactive food and feed ingredients and for valorization of side-streams from e.g., the fish processing industry. We present a novel approach for hydrolysate characterization that utilizes proteomics data for calculation of weighted mean peptide properties (length, molecular weight, and charge) and peptide-level abundance estimation. Using a novel bioinformatic approach for subsequent prediction of biofunctional properties of identified peptides, we are able to provide an unprecedented, in-depth characterization. The study further characterizes bulk emulsifying, foaming, and in vitro antioxidative properties of enzymatic hydrolysates derived from cod frame by application of Alcalase and Neutrase, individually and sequentially, as well as the influence of heat pre-treatment. All hydrolysates displayed comparable or higher emulsifying activity and stability than sodium caseinate. Heat-treatment significantly increased stability but showed a negative effect on the activity and degree of hydrolysis. Lower degrees of hydrolysis resulted in significantly higher chelating activity, while the opposite was observed for radical scavenging activity. Combining peptide abundance with bioinformatic prediction, we identified several peptides that are likely linked to the observed differences in bulk emulsifying properties. The study highlights the prospects of applying proteomics and bioinformatics for hydrolysate characterization and in food protein science.

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Proteolytic patterns and plasmin activity in ewes' milk as affected by somatic cell count and stage of lactation

Journal of Dairy Research ◽

10.1017/s0022029904000676 ◽

2005 ◽

Vol 72 (1) ◽

pp. 86-92 ◽

Cited By ~ 33

Author(s):

Marzia Albenzio ◽

Mariangela Caroprese ◽

Antonella Santillo ◽

Rosaria Marino ◽

Antonio Muscio ◽

...

Keyword(s):

Somatic Cell ◽

Cell Count ◽

Proteolytic Activity ◽

Somatic Cell Count ◽

Colorimetric Assay ◽

Sodium Caseinate ◽

Blood Stream ◽

Plasmin Activity ◽

Peptide Profile ◽

Milk Samples

A total of 120 milk samples were collected from Comisana ewes throughout lactation. The ewes were ranked into two somatic cell count (SCC) categories: normal milk (N Milk) with SCC lower than 5·00×105/ml and high somatic cell milk (HSC Milk) with SCC higher than 1·00×106/ml. Milk samples were analysed in triplicate for pH, fat and protein contents, renneting parameters, and plasmin and plasminogen activities. The peptide profile due to total proteolytic activity (endogenous and exogenous enzymes) on α- and β-CNs were determined using urea-PAGE on sodium caseinate (pH 8·0 and pH 5·0) incubated at 37 °C for 4 d after sampling. The peptide profile due to non-plasmin enzyme activities at pH 5·0 was also determined using urea-PAGE. Plasmin activity was higher in the HSC milk than in the N milk throughout the study period. A decrease in plasmin activity was observed in the N milk during mid-lactation, which was probably related to decrease in pH, and in the HSC milk during late lactation, which may be ascribed to an enhanced influx of plasmin inhibitors from the blood stream. Proteolytic patterns in Comisana ewe milk were mainly affected by plasmin activity that increased with the SCC in milk. Also non-plasmin proteolytic activity was strongly enhanced by elevated SCC and resulted in a higher degradation of α-casein than of β-casein. In general, plasmin activity did not increase with the advancement of lactation and exhibited a different trend in HSC and N milk, suggesting that physiological factors did not play a key role in regulating the plasminogen-plasmin system in ewes' milk. Plasmin activity, detected with the colorimetric assay was consistent with proteolytic activity on sodium caseinate shown in urea-PAGE electrophoregram.

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Correlation between the Degree of Hydrolysis and the Peptide Profile of Whey Protein Concentrate Hydrolysates: Effect of the Enzyme Type and Reaction Time

American Journal of Food Technology ◽

10.3923/ajft.2013.1.16 ◽

2012 ◽

Vol 8 (1) ◽

pp. 1-16 ◽

Cited By ~ 17

Author(s):

Harriman A. Morais ◽

Marialice P.C. Silve ◽

Viviane D.M. Silva ◽

Mauro R. Silva ◽

Ana Cristina S ◽

...

Keyword(s):

Reaction Time ◽

Whey Protein ◽

Whey Protein Concentrate ◽

Degree Of Hydrolysis ◽

Protein Concentrate ◽

Peptide Profile

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Degree of hydrolysis and peptide profile of whey proteins using pancreatin

Nutrire ◽

10.4322/nutrire.2013.026 ◽

2013 ◽

Vol 38 (3) ◽

pp. 278-290 ◽

Cited By ~ 7

Author(s):

Marialice Pinto Coelho Silvestre ◽

Harriman Aley Morais ◽

Viviane Dias Medeiros Silva ◽

Mauro Ramalho Silva

Keyword(s):

Whey Proteins ◽

Degree Of Hydrolysis ◽

Peptide Profile

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The level of bitterness and solubility of hydrolysates produced by controlled proteolysis of caseins

Journal of Dairy Research ◽

10.1017/s0022029900028831 ◽

1989 ◽

Vol 56 (3) ◽

pp. 375-379 ◽

Cited By ~ 15

Author(s):

G. E. Vegarud ◽

T. Langsrud

Keyword(s):

Proteolytic Enzymes ◽

Sodium Caseinate ◽

Degree Of Hydrolysis ◽

High Solubility ◽

Low Levels ◽

Casein Hydrolysates ◽

Rennet Casein

SummaryA series of hydrolysates from commercial sodium caseinate and rennet casein were prepared using different commercial proteolytic enzymes. Increasing degree of hydrolysis, DH, resulted in markedly changed solubility of the products. The level of bitterness varied with the enzyme used. Non-bitter hydrolysates of sodium caseinate were obtained with a DH of 10–12 and protein solubilities of 99%. The rennet casein hydrolysates exhibited low levels of bitterness and high solubility (90%), even at high DH values (DH = 55).

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Physicochemical and Nitrogen Solubility Properties ofBacillusProteinase Hydrolysates of Sodium Caseinate Incubated with Transglutaminase Pre- and Post-hydrolysis

Journal of Agricultural and Food Chemistry ◽

10.1021/jf011632n ◽

2002 ◽

Vol 50 (19) ◽

pp. 5429-5436 ◽

Cited By ~ 17

Author(s):

John Flanagan ◽

Richard J. FitzGerald

Keyword(s):

Nitrogen Solubility ◽

Sodium Caseinate

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Processing cottage cheese whey components for functional food production

Foods and raw materials ◽

10.21603/2308-4057-2020-1-52-59 ◽

2020 ◽

Vol 8 (1) ◽

pp. 52-59

Author(s):

Eugeniya Agarkova ◽

Alexandr Kruchinin ◽

Nikita Zolotaryov ◽

Nataliya Pryanichnikova ◽

Zinaida Belyakova ◽

...

Keyword(s):

Antioxidant Capacity ◽

Whey Protein ◽

Aspergillus Oryzae ◽

Functional Food ◽

Food Production ◽

Enzyme Preparation ◽

Cheese Whey ◽

Cottage Cheese ◽

Rational Approach ◽

Degree Of Hydrolysis

Introduction. The study offers a new rational approach to processing cottage cheese whey and using it as a highly nutritional functional ingredient in food production. We proposed a scientifically viable method for hydrolyzing cottage cheese whey with enzyme preparations of acid proteases from Aspergillus oryzae with an activity of 400 units/g and a pH range of 3.0 to 5.0. Study objects and methods. Pre-concentrated whey was enzymatically hydrolyzed at 30°C, 40°C, and 50°C for 60 to 180 min (pH 4.6). Non-hydrolyzed whey protein concentrates were used as a control. The amount of enzyme preparation was determined by calculation. All hydrolysate samples showed an increase in active acidity compared to the control samples. Further, we conducted a full-factor experiment with three levels of variation. The input parameters included temperature, duration of hydrolysis, and a substrate-enzyme ratio; the output parameters were the degree of hydrolysis and antioxidant capacity. Results and discussion. The experiment showed the following optimal parameters for hydrolyzing cottage cheese whey proteins with the enzyme preparation of proteases produced by Aspergillus oryzae: temperature – 46.4°C; duration – 180 min; and the amount of enzyme preparation – 9.5% of the protein content. The antioxidant capacity was 7.51 TE mmol/L and the degree of hydrolysis was 17.96%. Conclusion. Due to its proven antioxidant capacity, the whey protein hydrolysate obtained in the study can be used as a functional food ingredient.

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In Vitro Antioxidant and Antihypertensive Activity of Edible Insects Flours (Mealworm and Grasshopper) Fermented with Lactococcus lactis Strains

Fermentation ◽

10.3390/fermentation7030153 ◽

2021 ◽

Vol 7 (3) ◽

pp. 153

Author(s):

Adilene Mendoza-Salazar ◽

Lourdes Santiago-López ◽

María J. Torres-Llanez ◽

Adrián Hernández-Mendoza ◽

Belinda Vallejo-Cordoba ◽

...

Keyword(s):

Antioxidant Activity ◽

Lactococcus Lactis ◽

Antihypertensive Activity ◽

Angiotensin Converting Enzyme Inhibition ◽

Degree Of Hydrolysis ◽

Buffer Solution ◽

Solution Ph ◽

Total Polyphenol ◽

Peptide Profile ◽

Edible Insect

The objective of the present study was to evaluate the potential antioxidant and angiotensin converting enzyme inhibition (ACEI) activity of edible insect flours fermented with Lactococcus lactis strains. For the fermentation, mealworm and grasshoppers flours were dissolved (0.5% w/v) in buffer solution (pH 7.0) and individually inoculated (3%) with Lactococcus lactis strains (NRRL B-50571, NRRL B-50572). The samples were incubated for 72 h at 30 °C, and the pH was recorded. The degree of hydrolysis (DH) and protein content were determined. The total polyphenol compounds, antioxidant activity (ABTS, DPPH, ORAC, and FRAP), and ACEI of the <3 kDa fractions were analyzed. The pH of the fermented samples decreased to 3.5–3.9 (p < 0.05). The fermented grasshopper flour showed an increased DH (0.42%) and overall higher total polyphenol content (8.23 mg Gallic Acid Equivalent/mL). In general, the highest antioxidant activity was for the grasshopper fractions fermented for 24 h by Lactococcus lactis NRRL B-50572, which also showed 23.47% ACEI inhibition with an IC50 of 0.97 mg/mL. The peptide profile obtained increased after fermentation, being higher for the mealworm flour fermented sample. This study presents, for the first time, the use of specific strains of Lactococus lactis for fermenting edible insect-derived products in the production of bioactive compounds with potential antioxidant and antihypertensive activity.

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Increasing the Yield of Soluble Mustard Protein Isolate

Journal of Food Research ◽

10.5539/jfr.v4n6p124 ◽

2015 ◽

Vol 4 (6) ◽

pp. 124

Author(s):

L. Karina Lorenzo ◽

Levente L. Diosady

Keyword(s):

Enzyme Preparation ◽

Nitrogen Solubility ◽

Protein Isolate ◽

Cross Linking ◽

Protein Hydrolysis ◽

C Protein ◽

Food Applications ◽

Ph Range ◽

Colloid Formation ◽

Mustard Protein

The objective of this study was to investigate methods for improving the yield of acid soluble mustard protein isolate (SPI) by solubilizing isoelectrically precipitated protein isolate (PPI). The SPI is more valuable, as it can be used in unique food applications. Four treatments were tested in the acidic pH range: Alcalase hydrolysis; transglutaminase cross-linking; salting in with NaCl, Na5P3O10, and (NaPO3)6; and protective colloid formation with pectin. The effectiveness of each treatment was determined by measuring the increase in nitrogen solubility (AOCS-Ba11-65). Alcalase hydrolysis improved PPI solubility evenly in the 2.5-3.5 pH range, effectively eliminating the solubility minimum near the isoelectric point. At pH 3, the hydrolysis treatment increased solubility from ~20% to a maximum of ~70% (0.04 g of enzyme preparation / g PPI, 2 h, pH 8.5, 50-55oC). Protein hydrolysis during isolate production could increase the yield of SPI from 0.16 to 0.75 kg per kilogram of mustard protein.

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