Production Of Protein Hydrolysates Research Articles

Production and characteristics of protein hydrolysate from tra fish (Pangasius hypophthalmus)

The goal of this study was investigate effects of pH and temperature on the production of tra fish protein hydrolysate (FPH) fish protein concentrate (FPC) using alcalase and characterization of FPHs. Temperature of 55 o C and pH 8.5 were the best condition for highest degree of hydrolysis (around 55%) after 60 min of reaction. Under hydrolysis, protein solubility was significantly increased from 4% to >90%. FPC contained fractions of 50, 40, 37, 15, 12, and 11×10 3 g/mol. After 15 or 120 min of enzymatic reaction, molecular weight was degraded to 4.86 and 4.31 (x10 3 g/mol) of FPH15 or FPH120 samples. Due to the increasing of protein solubility during the hydrolysis, water holding capacity reduced from 2.81 ml/g (FPC) to FPH120>FPC and FPH120>FPH15>FPC. Furthermore, foam activity index and foam stability index of samples were arranged as FPH15>FPH120> FPC. At pH 4.0, all these indexes were lowest. Generally, under enzymatic hydrolysis, FPH could be possessed of different molecular weights and physico-chemical properties. Thus, FPH could reach various demand of industrial applications.

Enzymatic Production of Sarotherodon galillaeus Muscle Protein Hydrolysates and Assessment of its Alpha-Amylase Inhibitory and Antioxidant Potential

The study was carried out to evaluate the antioxidant activity and α-amylase activity inhibitory potential of Sarotherodon galillaeus muscle protein hydrolysates. Sarotherodon galillaeus muscle protein isolate was hydrolysed with three digestive proteases namely trypsin, chymotrypsin and pepsin. Degree of hydrolysis was determined. The antioxidative potential of the hydrolysates was investigated using DPPH radical scavenging, ferric reducing power, hydrogen peroxide scavenging and metal chelating activity. The ability of the hydrolysates to inhibit the activity of sugar-hydrolysing enzyme was also evaluated. Highest degree of hydrolysis was obtained with pepsin (55.86%) followed by trypsin (47.11%) and chymotrypsin (42.36%) after 6 hrs of hydrolysis. The half maximal inhibitory concentration (IC50) of hydrolysates produced by trypsin, chymotrypsin and pepsin for DPPH radical scavenging activity were 1.26 ± 0.95, 0.98 ± 0.07 and 1.18 ± 0.34 mg/ml respectively. Trypsin-produced hydrolysates displayed highest metal chelating activity and ferric reducing power while hydrolysates obtained with pepsin showed highest hydrogen peroxide scavenging activity. Amylase activity inhibitory potential of all the hydrolysates was low with pepsin-produced hydrolysates shown highest inhibitory effect of 18.46 ± 1.51%. The hydrolysates showed antioxidative potential that can be used in prevention of food oxidation.

ANTIOXIDANT ACTIVITY OF PROTEIN HYDROLYSATE PRODUCED FROM TUNA EYE (Thunnus sp.) BY ENZYMATIC HYDROLYSIS

Tuna (Thunnus sp.) by-products from frozen loin and canning industry especially the eye is rich in proteins and in lipids consisting of polyunsaturated fatty acids (PUFA). That requires protective agent (antioxidant) to inhibit the oxidation naturally present and predicted to be protein peptides. Enzymatic hydrolysis of protein is an appropriate method to produce bioactive peptide with such nutraceutical/pharmaceutical function such as an antioxidant peptide. This study aimed to produce protein hydrolysate having a function as anwith an antioxidant activity from eye of tuna through enzymatic hydrolysis and determining the antioxidant activity by DPPH methods. Protein soluble content of tuna’s eye protein hydrolysate (TEPH) ranged from 59.98±0.130 to 94.90±0.002%. The degree of hydrolysis (DH) of TEPH was about 9.10±0.28 to 16.14±0.09%. The highest inhibition of DPPH radical scavenging activity was 93.57±0.05% (at 5 mg/mL) was obtained with a DH of 11.35±0.002% at the concentration 0.1% of papain for 6 hours hydrolysis. The IC50 value of was 1.08±0.008 mg/mL

Protein hydrolysate from salmon frames: Production, characteristics and antioxidative activity.

Protein hydrolysates from two forms of salmon frames named "chunk" and "mince" were produced and characterized. Both samples were subjected to hydrolysis using alcalase and papain at 1%-3% (w/w protein) for 0-240min. Hydrolysate prepared with either protease at 3% for 180min had the solid yield of 24.05%-26.39%. Hydrolysates contained 79.20%-82.01% proteins, 6.03%-6.34% fat, 9.81%-11.09% ash, and 4.02%-5.80% moisture. Amino acid profile showed that all hydrolysates had glutamic acid/glutamine (113.45-117.56mg/g sample), glycine (77.86-86.18mg/g sample), aspartic acid/asparagine (76.04-78.67mg/g sample), lysine (61.97-65.99mg/g sample), and leucine (54.30-57.31mg/g sample) as the predominant amino acids. The size distributions determined by gel filtration chromatography varied, depending on proteases and the form of frame used for the hydrolysis. Different hydrolysates showed varying antioxidant capacities. Thus, protein hydrolysates from salmon frame could be used as a nutritive supplement in the protein deficient foods. PRACTICAL APPLICATIONS: Frames of salmon are by-products from salmon fish processing industries. The frames contained the remaining meat, hence they can be used for the preparation of protein hydrolysates. Generally, hydrolysates from fish by-products have been regarded as a promising food supplement, because they are rich in amino acids. Additionally, hydrolysates possess antioxidant activity, which is of health benefit. To produce the hydrolysate with less time consumption, the use of frame chunk instead of minced frame can be of better choice. Thus, frame of salmon, especially in chunk form, could be used as a raw material for production of protein hydrolysate using alcalase. The hydrolysate produced from salmon frame could serve as an alternative nutritive supplement to tackle the nutrition inadequacies in foods.

Physicochemical, antioxidant, calcium binding, and angiotensin converting enzyme inhibitory properties of hydrolyzed tomato seed proteins.

The objective of this was to determine the impact of enzymatic hydrolysis on the multifunctionality of tomato seed protein hydrolysates (TSPH) and their physicochemical properties. The enzymatic hydrolysis was performed using alcalase and two factors response surface methodology. The best conditions were 131.4min and 3% enzyme/substrate (E/S) for antioxidant activity; 174.5min and 2.93% E/S for angiotensin-converting enzyme (ACE) inhibition; and 66.79min and 2.27% E/S for the calcium binding. Antioxidant and ACE hydrolysates were characterized by higher solubility, zeta potential, and thermal stability while properties of the calcium binding hydrolysate were only minimally affected by the enzymatic hydrolysis. Gel electrophoresis showed that molecular weights of polypeptides in the calcium binding TSPH were higher compared to those in ACE and antioxidant TSPHs. This was due to the low degree of hydrolysis of the calcium binding hydrolysate. PRACTICAL APPLICATIONS: Nowadays, different protein sources are used to produce protein hydrolysates containing bioactive peptides that can help alleviate oxidation of foods, oxidative stress, and chronic conditions (e.g., hypertension, diabetes, cardiovascular disorder). Hydrolyzed proteins also have the potential to increase mineral absorption through the formation of mineral-binding complexes. Biological activities of proteins and peptides from tomato processing byproduct (i.e., pomace) have received until now little attention. The determination of physicochemical properties and biological activities of the hydrolyzed proteins has application in the formulation of value-added food products for the reduction of oxidative stress and risks of developing chronic diseases. In addition, there will be a reduction of pomace waste generated by the tomato processing industry.

Statistical optimization of enzymatic hydrolysis of rice bran protein concentrate for enhanced hydrolysate production by papain

Protein hydrolysates having higher degree of hydrolysis (DH) not only exhibit biological activities but also better functionalities than hydrolysates having low DH or intact proteins. The enzymatic hydrolysis of rice bran protein concentrate with papain to obtain protein hydrolysates was optimized using response surface methodology. A central composite rotatable design was used to study the influence of hydrolysis conditions, namely temperature (40–60°C), time (60–180 min), pH (6.0–7.0) and enzyme to substrate ratio (0.02–0.04 w/w) on DH. The experimental values of DH ranged between 14.08 and 33.09%. The second order polynomial model obtained for DH revealed a non-significant value of lack of fit test and coefficient of determination (R2) of 0.96. The optimum conditions for obtaining the higher DH using papain were pH 7.0, temperature 50°C, hydrolysis time 150 min and enzyme to substrate ratio 0.035 (w/w). Under these conditions, the experimental DH of hydrolysate was 34%, which agreed closely with the predicted value (33%). The peptide profile revealed that papain was a potential enzyme to produce protein hydrolysate with more small peptides as compared to free amino acids.

Improving antioxidant activity of black bean protein by hydrolysis with protease combinations

SummaryThis study explored the use of a simplex centroid design to produce protein hydrolysates with antioxidant properties using Alcalase® 2.4L, Flavourzyme® 500L and Neutrase® 0.8L. Proteases kinetic parameters and the ultrafiltration of protein hydrolysates were also investigated. The highest antioxidant activity, in the studied conditions, was reached when the mixture of Alcalase® 2.4L and Flavourzyme® 500L was used in the hydrolysates production. The antioxidant power of the black bean proteins, measured by the total antioxidant capacity and reducing power assay, increased after hydrolysis by 31% and 70%, respectively. The black bean proteins hydrolysates fractions (3–30 kDa) showed an antioxidant activity decrease along with a reduction in molecular weight, demonstrating that a set of varied molecular weight peptides was responsible for the antioxidant characteristics of black bean protein hydrolysates.

Low-cost culture medium for the production of proteases by Bacillus mojavensis SA and their potential use for the preparation of antioxidant protein hydrolysate from meat sausage by-products

The present study aims to maximize proteases production by Bacillus mojavensis SA strain and their use to produce bioactive protein hydrolysates from a meat by-product. The production of SA bacteria proteases was maximized using a culture medium based on wheat bran, which offer an advantage in minimizing the production cost and enhancing the enzyme activity by using agro-industrial wastes. The composition of media and cultural conditions for optimal proteases production by B. mojavensis SA strain were investigated. A successful and significant improvement of the alkaline proteases production (four folds) by the SA strain was achieved using the medium composed of (g/l): wheat bran, 50.0; KH2PO4, 0.5; K2HPO4, 0.5; CaCl2, 2.0; pH 6.0, where the growth conditions were monitored at 37 °C with an agitation speed of 200 rpm. Interestingly, the enzyme preparation of B. mojavensis was applied for the preparation of protein hydrolysates from a meat by-product. Hydrolysis was carried out for 180 min at pH 12.0. The resulting hydrolysate displayed an important antioxidant activity as evaluated by the radical scavenging capacity, the reducing power, and the β-carotene bleaching inhibition. The present study showed the high proteases’ producing level by B. mojavensis SA strain in a low-cost fermentation medium (wheat bran) and their potential use in the production of bioactive protein hydrolysate from meat by-products.

Investigation of potent odorants generated during the production of whey protein hydrolysates.

Whey protein hydrolysates (WPH) were prepared from whey protein concentrate (WPC) by partial or extensive hydrolysis. WPC and two WPH types were characterized by a descriptive sensory analysis. WPC exhibited a "milky", "sweet", and "mild milk fat" odor, whereas extensively hydrolyzed whey protein (E-WPH) demonstrated "cheese-like", "acrid odor", and "sour" olfactory characteristics. In E-WPH, 3-(methylthio)-propanal, phenylacetaldehyde, and 3-methylbutanoic acid were specifically detected with high flavor dilution factors upon aroma extract dilution analysis. These odor-active compounds likely contributed to the olfactory characteristics of E-WPH, and their concentration increased together with that of their respective putative precursors: free methionine, phenylalanine, and leucine.

Scaling up process for fish peptone production

Fish peptone is one of protein hydrolysate products that can be used as nitrogen source for microbial growth. Peptone can be made from marine fish whereas protease enzyme was made from either papaya extract or commercial papain. Fish peptone is not only an eco-friendly approach but also can produce economic materials used in formulation of culture media. The objective of this research was to optimise the scaling up process of fish peptone production. An amount of 200 g of clean fish marine was blended with 1 L of water and added with 0.2 % of commercial papain at pH 8. Peptone production was carried out in a 1 L glass reactor with agitation (50, 100, 150, and 200 rpm) for 1-8 h. Fish slurry was then incubated at 50°C for 8 h. Fish protein hydrolysate was spray-dried with inlet and outlet temperature of 90°C and 80°C, respectively, at flow rate of 200 mL/h. Results revealed that the best condition for hydrolysis was shown by protein soluble content of 17.1 mg/L, α-amino content of 147 mM, and degree of hydrolysis of 8.3% which was achieved by agitation at 100 rpm for 8 h reaction. The yield of dried peptone product was 11.9 % (w/v). This peptone powder had higher soluble protein and α-amino content (442.3 mg/g, 124 mM) than the commercial peptone (258.5 mg/g, 65 mM). The microbial growth test of fish peptone products showed similar effectiveness compared with commercial bacterial peptone for the growth of Escherichia coli and Streptococcus aureus.

Antioxidant Properties of Flaxseed Protein Hydrolysates: Influence of Hydrolytic Enzyme Concentration and Peptide Size

AbstractFlaxseed is an emerging source of protein raw materials for bioactive protein hydrolysate production; however, there is insufficient information on the structural properties of its antioxidative peptides. The aim of this work was to determine the peptide composition and relationships to antioxidant properties of flaxseed protein hydrolysates (FPH) obtained using two protease (thermoase) concentrations (2.5 and 3.0%). The FPH were passed through ultrafiltration membranes to yield fractions with <1, 1–3, 3–5, and 5–10 kDa peptide sizes, which were also tested for antioxidant properties. Mass spectrometry data yielded several peptide sequences located within the flaxseed conlinin, but a lesser number was found in the 3.0% thermoase‐FPH. 2,2‐Diphenyl‐1‐picrylhydrazyl radical scavenging was lowest (54%) for the 3.0% thermoase‐FPH while membrane separation enhanced the values up to 65% for the <1 kDa peptides. Metal chelation and iron‐reducing ability were also significantly (P < 0.05) enhanced after membrane fractionation. In contrast, the hydroxyl radical scavenging ability was unchanged (2.5% thermoase‐FPH) or significantly (P < 0.05) diminished (3.0% thermoase‐FPH) after membrane fractionation, except for the 5–10 kDa peptides that had notably (P < 0.05) higher values. We conclude that the <1 and 1–3 kDa peptides may be useful agents to suppress oxidative stress within living tissues while the 3–5 and 5–10 kDa could be suitable preservatives to enhance food shelf life.

RESEARCH ON THE SUITABLE PARAMETERS FOR HYDROLYSIS REACTION OF RED MEAT OF STRIPED TUNA (SARDA ORIENTALIS) BY USING COMMERCIAL PROTAMEX

Red meat of Sarda orientalis is one of by-products of tuna processing industry, the source of Sarda orientalis red meat is abundant in Vietnam. Red meat of Sarda orientalis contained 27 % of protein and 0.53 % of lipid, thus, it was classified into lean fish meat and a perfect resource for protein hydrolysate production. The purpose of this research is to hydrolyze Sarda orientalis red meat by using protamex enzyme to obtain protein hydrolysates. The suitable parameters for hydrolysis reaction were reaction time of 4 h, ratio of Protamex enzyme to red meat of 0.4 % (based on the weight of red meat, w/w), and reaction temperature of 55 °C with the maximal value of degree of hydrolysis of 7.03 %. The highest degree of hydrolysis in this research can compare to previous researches.

Yield, physicochemical, and antioxidant properties of Atlantic salmon visceral hydrolysate: Comparison of lactic acid bacterial fermentation with Flavourzyme proteolysis and formic acid treatment

This study compares a cost-effective microbial method, involving lactic acid (LA) fermentation, with the conventional formic acid (FA) treatment and Flavourzyme (FL) enzymatic hydrolysis for valorizing Atlantic salmon processing waste (Viscera). LA and FA processing approaches relied on production (LA) or addition (FA) of organic acids to lower the pH and activate the inherent proteases in these tissues, whereas FL processing was carried out at the optimum conditions for Flavourzyme (37 °C, pH 7.0). Highest protein hydrolysate fraction recovery (∼57%) was seen in LA fermentation, while FA processing resulted in the highest protein levels (∼87%) in the recovered hydrolysate fraction and lowest residual fraction (∼4%). In general, FL processing demonstrated higher Fe(II) chelation (73%) and ferric reducing capacity (27 mM glutathione equivalent) whereas LA-hydrolysates showed enhanced sacrificial antioxidant properties. Findings from this study would have implications on the application and production of salmon-based protein hydrolysates, especially, from the perspective of their functionality. Practical applications Peptides derived from natural sources such as fish products have been extensively reported to exert antioxidant activity that mitigates oxidative stress and providing beneficial effects to human health as well as food storage. Several sources of proteins are being explored for the production of biologically active peptides. Utilizing by-product streams such as fish processing waste is especially attractive as these are cheaper and rich sources of protein. Therefore, utilizing marine bioprocessing waste for the production food-grade protein hydrolysates is promising, especially in Atlantic Canada.

RELATIONSHIP BETWEEN BITTERNESS OF BREWER’S YEAST HYDROLYSATE AND HYDROPHOBIC AMINO ACID CONTENT

Brewer’s yeast spent, obtained after the main fermentation stage, is a rich- in-protein source(protein content accounts for 48 - 50 % dry matter). In order to use efficiently this source, it washydrolysed by different methods. Protein hydrolysate products are normally mixtures of peptidesand amino acids. Protein hydrolysates have a wide range of applications in food. It can be usedas emulsifying agents in a number of applications such as salad dressings, spreads, ice cream,coffee whitener, cracker, and meat products like sausages. However, bitterness in hydrolysates isone of the major undesirable aspects for various applications in food processing. In this study,we used enzymatic mixture alcalase and flavourzyme, yeast treatment methods to hydrolysebrewer’s yeast. The hydrolysate and fractions of protein hydrolysate obtained after filtration with10 kDa and 3 kDa filters were used for determination of bitterness and hydrophobic amino acidscontent. The bitter taste of hydrolysate was determined by sensory method (using quininestandard) and amino acid content was analysed by HPLC method. The result showed the closerelationship between bitter taste and hydrophobic amino acid content. The bitter taste of proteinhydrolysate was reduced as the hydrophobic amino acid content decreased. When the bitter taste(equivalent to quinine concentration) decreased from 16.25 μmol/l to 3.59 μmol/l, the totalcontent of hydrophobic amino acids in protein hydrolysate reduced from 1653 μg/ml to 932μg/ml.

Comparison of the influence of pH on the selectivity of free and immobilized trypsin for β-lactoglobulin hydrolysis

Although immobilized trypsin is a viable alternative to the free one in solution for producing protein hydrolysates, the change of selectivity introduced by immobilization is unclear. In this study, we compared the selectivity of free and immobilized trypsin towards different cleavage sites of β-lactoglobulin (β-Lg) with a focus on the impact of environmental pH. Both free and immobilized trypsin exhibited greater accessibility to native β-Lg at elevated pH (from pH 7.2 to 8.7). Additionally, free trypsin preferred to attack cleavage sites located at the C-terminus at pH 7.8, whereas an opposite preference for the N-terminus was observed at pH 8.7. Regarding the immobilized trypsin, the pH did not significantly influence its preference for the C- or N-terminus. Generally, immobilization of trypsin resulted in more focused cleavage within its specificity during the initial stage of hydrolysis, and some peptides were formed more rapidly by the immobilized trypsin.

Kinetics of enzymatic reactions in the production of fish protein hydrolysates

ABSTRACTThe kinetic laws of enzyme hydrolysis of waste products (heads, fins, bones, and muscle tissue) of the Atlantic cod Gadus morhua have been investigated. An enzyme specimen hepatopancreatine was used for the hydrolysis reaction (proteolysis). A kinetic model describing the mechanism of enzyme hydrolysis of fish proteins as a second-order reaction was developed. The proteolysis model was based on the existence of easily hydrolysed and hydrolysis-resistant protein fractions. Autolysis of the enzyme was also taken into consideration. In the context of the developed kinetic model, the second-order rate constants of the intermediate stages of proteolysis were calculated.

Conversion of chicken viscera into protein hydrolysate for palatant production

Flavor and aroma cannot be separated in food industries. The manipulation of the ingredients to increase the taste has started since early civilization. Common example of the earliest application of ingredients to increase the savory taste of the food is soy sauce production. The process of digging out the aroma and flavor includes frying, stewing, grilling as well as fermenting. Nowadays, much complex savory system in food can be achieved through several processes such as Maillard reaction and protein hydrolysate production. The food produced will not only be tasty but healthy. This concept was first focusing on human food production. However, the increasing number of pet owner for about 10% annually since 2008 as reported by US Pet Owner Society demands the same concept to be implemented in pet food industries. Since most pet owners considered their pets as part of their family, the pet food source from 4D (dead, dying, disable and diseased) animals are unacceptable. Therefore, the pet food industries are competing with human food industries to get the clean source of meat for food as well for flavor production. An alternative to overcome this is by utilizing the viscera of the slaughtered chicken for palatant or flavor production. Aside from meat, bones and feathers, viscera are also abundantly being considered as waste and were not fully utilized. They are rich in protein. The protein can be recovered using suitable process and later turned into a value-added product such as flavor. There is a study available for converting the protein waste into protein hydrolysate via chemical and physical technique. However, only few reports are available for the conversion process via indigenous microbes and enzymes. It is believed that the indigenous microbes and enzymes (protease, peptinase) can be utilized for protein hydrolysate production which will later can be utilized as palatant. The palatant produced should have Sulphur-based aroma compound such as 2-methyl-3-furanthiol and 2-furfuryl-thiol which serve as precursors for chicken aroma.

Production of fish protein hydrolysate from silver catfish (Arius thalassinus)

Production of fish protein hydrolysate from silver catfish (Arius thalassinus)

Antioxidant and antimicrobial activity of ultra-filtered fractions of camel milk protein hydrolysates under in-vitro condition

Sequential ultra-filtration technique was used to fractionate camel milk protein hydrolysates products by 3 different proteolytic enzymes, viz. alcalase, α-chymotrypsin and papain. The protein fractions were assessed for antioxidant activities, viz. 2,2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), 2,2-diphenyl-1 picrylhydrazyl (DPPH) and ferric reducing antioxidant power assay (FRAP), and antimicrobial activity (inhibition zone assay). The whole hydrolysates recorded significantly higher inhibition activity in ABTS, DPPH and FRAP assay, whereas among fractions, F2 (1-5 kDa) and F3 (5-10 kDa) of all the 3 hydrolysates had higher activity. Higher antioxidant activities was also observed in α- chymotrypsin hydrolysates samples and its fractions followed by alcalase and papain. The zone of inhibition (mm) was also recorded higher for whole hydrolysates as compared to their fractions; however, different fractions had almost comparable antimicrobial effect. The protein hydrolyastes with alcalase and α- chymotrypsin recorded comperatively higher antimicrobial activity. The findings suggested that camel milk proteins could be valuable source to produce protein hydrolysates and ultra-filtration technique could also be used to get specific molecular weight peptides, however, for application in processed food or for direct human consumption, use of whole hydrolysates could be more beneficial and cost effective.

Production of Fish Protein Hydrolysates from Scyliorhinus canicula Discards with Antihypertensive and Antioxidant Activities by Enzymatic Hydrolysis and Mathematical Optimization Using Response Surface Methodology.

Fish discards are of major concern in new EU policies. Alternatives for the management of the new biomass that has to be landed is compulsory. The production of bioactive compounds from fish protein hydrolysates (FPH) has been explored in recent years. However, the viability of Scyliorhinus canicula discards, which might account for up to 90–100% of captures in mixed trawler, gillnet, and longline industrial fisheries, to produce FPH from the muscle with bioactivities has still not been studied in terms of the optimization of the experimental conditions to enhance its production. The effect of pH and temperature on the hydrolysis of the S. canicula muscle was mediated by three commercial proteases using response surface methodology. Temperatures of 64.6 °C and 60.8 °C and pHs of 9.40 and 8.90 were established as the best hydrolysis conditions for Alcalase and Esperase, respectively. Optimization of the best conditions for the maximization of antihypertensive and antioxidant activities was performed. Higher Angiotensin-converting enzyme (ACE) activity was found with Esperase. The pH optimum and temperature optimum for antioxidants were 55 °C/pH8.0 for ABTS/DPPH-Esperase, 63.1 °C/pH9.0 for DPPH-Alcalase, and 55 °C/pH9.0 for ABTS-Alcalase. No hydrolysis was detected when using Protamex.