Production Of Fish Protein Hydrolysates Research Articles

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

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

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.

Impact of microwave-assisted enzymatic hydrolysis on functional and antioxidant properties of rainbow trout Oncorhynchus mykiss by-products

Fishery by-products can be better utilized following enzymatic hydrolysis treatment to produce fish protein hydrolysates (FPH) with potentially enhanced interface-stabilizing properties (e.g. functionality). The production of FPH could be accelerated through the application of rapid heating methods [e.g. microwave-assisted heating (MW)] rather than slower conventional heating (CH) treatments. The objective of this study was to investigate the effects of microwave heating during enzymatic hydrolysis on the functionality and antioxidant properties of FPH. Trout by-products were hydrolyzed with Alcalase at an enzyme substrate ratio (E:S) of 0.5, 1.7, and 3.0% (w/v), respectively, for 3, 5 and 15 min using a microwave system (1200 W, 20% power with 50% duty cycle at 50–55 °C) and a conventional heating method (water bath at 50 °C). The degree of hydrolysis and protein solubility was higher (P < 0.05) for the MW-FPH than for the CH-FPH. MW-FPH at 5 min (0.5% E:S) demonstrated higher (P < 0.05) emulsifying activity and emulsion stability than CH-FPH with the same treatment. Foam capacity and stability were also greater (P < 0.05) for MW-FPH samples that were treated 15 min by microwave-assisted heating (0.5% E:S) when compared to CH. Overall, MW-FPH exhibited higher (P < 0.05) 2,2-diphenyl-1-picryhydrazyl and ferric ion reducing capacity than CH-FPH. We therefore conclude that microwave-assisted hydrolysis is an alternative method to produce FPH with improved solubility, emulsifying activity, foaming properties and antioxidant activity.

Fish Protein Hydrolysate Production by Acid and Enzymatic Hydrolysis

The objective of this study was to optimize the condition for fish protein hydrolysate production from minced by-catch fish by using hydrochloric acid and 2 protease enzymes; Papain and Alcalase. Taguchi design and Central Composite Design (CCD) were applied for experimental design to evaluate the degree of hydrolysis. Response surface methodology was performed in order to determine the optimal production conditions. The optimal condition for acid hydrolysis was 4 mol/L of hydrochloric acid at 100 o C for 90 minutes which yielded 50.70% degree of hydrolysis. For enzymatic hydrolysis, Alcalase is the most suitable protease enzyme for fish protein hydrolysate production. The optimal condition was 6 % (w/w) Alcalase concentration at the temperature of 61.23 o C and the reaction time of 27.36 minutes resulting in 88.9% of degree of hydrolysis. Amino acid profiles of fish protein hydrolysates hydrolyzed under the optimal condition were analyzed by HPLC and the results showed that fish protein hydrolyzed by Papain had highest nutritional properties. Glutamic acid had the highest percentage (16.35%) followed by Aspartic acid (10.41%) and Lysine (8.48%).

Protein hydrolysate waste of whitemouth croaker (Micropogonias furnieri) as a way of adding value to fish and reducing the environmental liabilities of the fishing industry

As an alternative use of natural marine resources, processing waste from whitemouth croaker was used as raw material for the production of fish protein hydrolysate (FPH). The biometric characterization of the raw material was determined, and the optimal conditions of the protease established using DH (degree of hydrolysis) as the criterion. Bromatological analyses were performed according to AOAC methods. The DH were determined by the TCA (trichloroacetic acid) method and expressed as a percentage of solubilized protein after hydrolysis compared to the total protein in raw material. The average yields of viscera, heads and carcasses were 7.4, 36.3 and 56.3%, respectively. The optimal conditions for enzyme action on the viscera were: 55°C, hydrolysis time of 90 min, and enzyme concentration of 0.4 mg mL -1 . For the muscle, the conditions established were: 45°C, 90 min, and 0.8 mg mL -1 . The dry matter of hydrolysate from viscera yielded 73.5% supernatant (soluble fraction) and 27.5% precipitate (insoluble fraction), while that from muscle yielded 45.7% and 54.3%, respectively. The hydrolysate from viscera presented 86.0% proteins, 0.4% lipids and 4.4% ashes in the soluble fraction, and 51.0% proteins, 25.8% lipids and 5.1% ashes in the insoluble fraction. The hydrolysate from muscle presented 86.0% proteins, 0.4% lipids and 9.0% ashes in the soluble fraction, and 87.0% proteins, 4.4% lipids and 2.4% ashes in the insoluble fraction. A lower degree of hydrolysis of hydrolysate from muscle than that produced from viscera was observed, demonstrating the different interactions between the enzyme and the residues studied.

Antioxidative Activitiy of Protein Hydrolysate from the Muscle of Common Kilka (Clupeonella cultriventris caspia) Prepared Using the Purified Trypsin from Common Kilka Intestine

ABSTRACTTrypsin from the intestine of common kilka (Clupeonella cultriventris caspia) was purified using ammonium sulfate precipitation (30–50% saturation), Sephadex G-75, and DEAE-cellulose chromatography with the purity of 30-fold and the yield of 12%. The molecular weight of trypsin was estimated to be 23.2 kDa based on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The trypsin had optimal activity at pH 8.0 and 60°C using N-α-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) as a substrate and showed high stability in the pH range of 7.0–10.0. It was stable up to 50°C. Soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysine-chloromethylketone (TLCK) significantly inhibited trypsin activity (p < 0.05). Protein hydrolysate from common kilka muscle with different degrees of hydrolysis (DHs; 20, 30, and 40%) was prepared using the purified trypsin, and antioxidative activities were determined. 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing antioxidant power, and ferrous chelating activity of hydrolysate increased with increasing DH up to 40% (p < 0.05). Therefore, trypsin from intestine of common kilka could be used as a processing aid for production of fish protein hydrolysate with antioxidative activity.

Functional, bioactive and antigenicity properties of blue whiting protein hydrolysates: effect of enzymatic treatment and degree of hydrolysis.

Fish discards represent an important under-utilisation of marine resources. This study evaluated the up-grading of the protein fraction of blue whiting (Micromesistius poutassou) discards by the production of fish protein hydrolysates (FPHs) exhibiting functional, antioxidant, angiotensin-I converting enzyme (ACE)-inhibitory and antigenicity properties. FPHs with low DH (4%) showed better emulsifying, foaming and oil binding capacities, particularly those obtained using only trypsin. FPHs with DH 4% exhibited also the stronger antioxidant activity, especially the one obtained using only subtilisin (IC50 = 1.36 mg protein mL-1 ). The presence of hydrophobic residues at the C-terminal of the FPH produced using subtilisin also led to the stronger ACE-inhibitory activity. However, FPHs with high DH (12%), which implies a higher proportion of short peptides, was required to enhance ACE-inhibition (IC50 = 172 µg protein mL-1 ). The antigenic levels of the FPH were also reduced with DH independently of the enzymatic treatment. Nevertheless, the highest degradation of fish allergens (e.g. parvalbumin) was also obtained when using only subtilisin. These results suggest that added-value products for food applications can be produced from the protein fraction of discards. © 2016 Society of Chemical Industry.

Production of Fish Hydrolysates Protein From Waste of Fish Carp (Cyprinus Carpio) by Enzymatic Hydrolysis

Fish Protein Hydrolysates (FPH) is the mixed products of polypeptide, dipeptides, and amino acid. It can be produced from materials that contained of protein by acid reaction, base reaction or enzymatic hydrolysis. The objectives of this study were to study the production of FPH from fish carp meat at post rigor phase and viscera by enzymatic hydrolysis, to determine the specific activity of papain enzyme, and to determine the solubility of FPH. Capacity of fish hydrolyzing can be identified by analyzing the content of dissolved total nitrogen (NTT) compared with nitrogen total ingredient (NTB) in order to get the value of total soluble nitrogen/total nitrogen material (NTT/NTB). The hydrolysis processes were carried out in 0,26% (w/v) papain, 60 οC for 3 hours. The result showed that the specific activity of papain enzyme was about 3.28 U/mg. Solubility of FPH by comparing NTT/NTB was about 0.29% (fish meat) and 0.40% (fish viscera). Proximate test of protein content of fish meat was 18.34 ± 0.04 (g/100 g); while viscera was about 0.95±0.04 (g/100 g). The result indicated that product waste of fish carp had potential as a major of source of FPH.

Cape hake protein hydrolysates prepared from alkaline solubilised proteins pre-treated with citric acid and calcium ions: Functional properties and ACE inhibitory activity

The production of fish protein hydrolysates (FPH) is a well-established technology for the upgrading of fish by-products. However, the presence of lipids in the FPH may have adverse effects on their quality. Thus, the objective of this study was to prepare and characterise hake protein hydrolysates (HPH) from alkaline solubilised Cape hake by-products proteins previously treated with CaCl2 and citric acid to remove phospholipids (method C). For comparison protein hydrolysates were also prepared directly from raw material (method A) and from the alkaline solubilised proteins (method B).The yields obtained in the preparation of hydrolysates by the methods B and C were considerably lower than that obtained by method A. The hydrolysates prepared by methods B and C had also lower degree of hydrolysis (DH) and protein content, but all HPH presented a well-balanced composition of essential amino acids.The methods B and C led to a high reduction of lipid content of hydrolysates (ca. 90%), however, only the method C allowed a considerable reduction of phospholipids content. HPH-A was visually lighter and HPH-C showed the highest solubility (94.3%), emulsifying activity index (11.7m2/g) and emulsifying stability (97.9%). The addition of the three HPH's to minced gilthead seabream increased its water holding capacity (15.8–19.2%). HPH's exhibited angiotensin converting enzyme (ACE) inhibitory activity at 1mgmL−1, indicating the presence of peptides with antihypertensive activity. The method C was effective to remove phospholipids and allow obtaining HPH with good functional properties.

Production of Fish Protein Hydrolysates by Acid and Enzymatic Hydrolysis

Production of Fish Protein Hydrolysates by Acid and Enzymatic Hydrolysis

Utilization of tilapia processing waste for the production of fish protein hydrolysate

Viscera and carcasses represent 60–70% of whole fish body weight, and this processing waste is a known source of bioactive molecules, including proteases. These enzymes can be employed in various biotechnological processes, such as, for preparing fish protein hydrolysate (FPH). Therefore, the objective of this study was to evaluate the use of processing waste from Nile tilapia (Oreochromis niloticus) as a source of protein and proteases to produce FPH. Three FPH production conditions were evaluated: two conditions used autolysis with enzymes extracted from the tilapia intestine at different concentrations (FPH100, 100mg of tissue/mL and FPH600, 600mg of tissue/mL) and the third used 0.5% (v/v) Alcalase (FPHcom), a commercial protease preparation. Protein, amino acids and fatty acids content were calculated as DM basis. After a 4-h reaction, maximum hydrolysis percentages from FPHcom, FPH100, and FPH600 systems were 34.73±1.44%, 29.21±0.79%, and 41.66±1.33%, respectively. The protein content in the resulting FPS was 584.8g/kg, 492.3g/kg, and 508.2g/kg for FPHcom, FPH100, and FPH600, respectively. Methionine and lysine were found at levels of 32.0 and 77.0g/kg (FPHcom), 31.0 and 64.0g/kg (FPH100), and 33.0 and 69.0g/kg (FPH600), respectively. Polyunsaturated fatty acid contents of FPHcom, FPH100, and FPH600 were 101.0g/kg, 138.0g/kg, and 70g/kg, respectively, with a predominance of linoleic acid (C18:2n−6). Amino acid composition, lipid profile, and amino acid score suggested that all of the experimental FPHs could be employed as a protein source in diets for aquatic organisms and other farmed animals.

Production and fractionation of tuna by-product protein hydrolysate by ultrafiltration and nanofiltration: Impact on interesting peptides fractions and nutritional properties

The production of fish protein hydrolysates (FPH) is a promising route to add value to fish by-products due to their potential application as a source of interest peptide fractions. Using Alcalase for hydrolysis of tuna dark muscle by-product, the influence of enzyme/substrate ratio and hydrolysis time on the rate of interest peptide fractions (1–4kDa) was studied. The rate of this fraction obtained under optimized conditions (temperature 55°C, pH8.5, enzyme/substrate ratio of 1% and 60min of hydrolysis reaction) was 26%. The performance of the combined process associating ultrafiltration and nanofiltration membranes was evaluated for fractionation of produced hydrolysate in order to isolate fractions enriched in peptides of specific molecular weight (MW). The interest peptide fraction (1–4kDa) was isolated and the positive effect of diafiltration on peptide purification process was underlined. The peptide fractions produced have a high nutritional quality which can be used in human nutrition and they have a potential for applications in aquaculture diets.

Functional and antioxidative properties of protein hydrolysates from Cape hake by‐products prepared by three different methodologies

The production of fish protein hydrolysates (FPH) is a convenient technology for upgrading fish by-products. The aim of this work was to study three different methods of FPH preparation from Cape hake by-products to improve yield and quality. Functional and antioxidative properties of all FPHs were determined. The protein content of hake FPH was in the range 807-860 g kg(-1) and the degree of hydrolysis was between 19% and 22%. The maximum yield (71.9%) was achieved by methodology B but the hydrolysate was darker. The peptide profile of all FPHs was very similar. FPH prepared by methodology C had significantly higher emulsifying activity index and hydrolysate prepared by methodology B had the highest foaming capacity. The solubility of FPH was in the range 71-76% and increased the water-holding capacity of minced fish by about 9%. The fractionation of FPH obtained by methodologies A and B allowed concentrating peptides with higher radical scavenging activity and reducing power. The properties of the FPH prepared indicated that they can be used in food systems as natural additives, particularly to improve their water-holding capacity.

Fish protein hydrolysate production from sardine solid waste by crude pepsin enzymatic hydrolysis in a bioreactor coupled to an ultrafiltration unit

The aims of the study were to optimize the production a fish protein hydrolysate (FPH) by enzymatic hydrolysis of sardine solid waste using crude pepsin, and to scale up the process in a bioreactor coupled to an ultrafiltration unit for product recovery. Results showed that the crude pepsin prepared by autolysis of the mucous membranes of a sheep stomach at optimal conditions (i. e. pH=1.5–2 and incubation time of 6h) could be satisfactory used for the enzymatic hydrolysis of fish solid waste. The optimal conditions for enzymatic reaction were: temperature 48°C, and pH 1.5. The scale up of the enzymatic hydrolysis and the coupling of the reactor an ultrafiltration unit to concentrate the hydrolysate gave good results with a rejection coefficient for the protein hydrolysate product in the range of 90%. The volumetric concentration factor was 2.5, with a permeate flux of 200Lm−2bar−1. However, the results also suggest that the ultrafiltration product concentration process may be operating beyond the critical flux at which point irreversible membrane fouling occurs.

Enzymatic Hydrolysis of Fish Frames Using Pilot Plant Scale Systems

Papain was used to hydrolyse fish frames under controlled conditions at a batch-pilot plant scale- process, for the production of fish protein hydrolysates (FPH). Mass balance calculations were carried out so that the rate of hydrolysis, rate of protein solubilisation and yields could be estimated. Almost complete hydrolysis could be achieved in 1 hour, at 40oC, with no pH adjustment, at 0.5% (5 g.kg-1) enzyme to substrate ratio (E/S, were S is Kjeldahl protein) using whole fish frames (including heads and flaps). This was achieved both with the addition of water (1/1 to 2/1 frames/water) but more importantly from commercial considerations without the initial addition of water (after mincing of the fish material). The degree of protein solubilisation ranged between 71% - 86% w/w. Four different processes are described, namely: 1) a soluble spray-dried FPH powder 2) a liquid FPH 3) a partly soluble, spray dried FPH powder and 4) a crude, drum-dried protein for animal consumption. The amino acid profile of the FPH was identical to that of the parent substrate (fish frames).

Endogenous Proteases in Pacific Whiting (Merluccius productus) Muscle as A Processing Aid in Functional Fish Protein Hydrolysate Production

A fish protein hydrolysate (FPH) from Pacific whiting (Merluccius productus) muscle was produced by autolysis of a minced homogenate (8% protein) at pH 7.0 and 60°C, where maximum endogenous proteolytic activity was detected. FPH production was controlled by the pH stat method, yielding a 4.43% degree of hydrolysis after 1 h of autolysis. Upon autolytic processing, 28.9 ± 0.7% of the total protein was found in the soluble fraction. FPH was 100% soluble at pH 7.0 and 10.0 and was less soluble at pH 4 (82.5%, P ≤ 0.05). FPH showed better emulsifying properties than sodium caseinate (SCA) at pH 4.0 (P ≤ 0.05), but had a lower foaming capacity (P ≤ 0.05) than bovine albumin (BSA) at all evaluated pHs. FPH foaming capacity was not affected by pH, however, foam stability was equal or better than that of BSA, especially at pH 4.0 (P ≤ 0.05). These results suggest the possibility of producing FPH with similar or better functional properties than those of functional ingredients, such as SCA and BSA. Furthermore, the data presented support our hypothesis that the high proteolytic activity in Pacific whiting could be used as an advantage in fish protein hydrolysate production or as a processing aid where protein hydrolysis is required.

Pacific whiting ( Merluccius productus) underutilization in the Gulf of California: Muscle autolytic activity characterization

In México’s Northwest coast, Pacific whiting ( Merluccius productus) is considered an under-utilized species because of its high tendency to become parasitized, thus promoting a high proteolytic activity present in muscle tissue. Sample fish for study were separated in lots by degree of parasitism in “apparent” parasitized (APP) and advanced parasitized (ADP). Thus, pH and temperature conditions of mayor endogenous proteolytic activity in muscle were determined. Maximum activity was detected at 50 °C (pH 3.5–4.0) and at 60 °C (pH 6.75–7.0). Parasitism degree had a significant effect in enzyme activity at acidic pH ( p < 0.05) being higher in APP at low temperature (30 °C). Higher temperatures (40–50 °C) favored ( p < 0.05) activity in ADP muscle (same pH range) with the highest ( p < 0.05) observed at 50 °C at pH 3.5. No much difference was observed at pH 7.0–8.0. Results suggest that pH around physiological conditions at 60 °C could be used as an advantage in fish protein hydrolysate production or as processing aid where a protein hydrolysis is required.

Production of Fish Protein Hydrolysates with Bacterial Proteases; Yield and Nutritional Value

ABSTRACTThree bacterial proteases were selected for their activity on casein per unit cost. Conditions for inactivation were determined. Fish protein was hydrolyzed without water addition. Protein was solubilized faster with Pescalase 560 than with HT‐200 or Protease N. Increase of protease concentration was associated with a quadratic increase in soluble nitrogen. Yields of 13 to 15% in terms of whole fish were obtained at the pilot plant. Products contained 83 to 86% protein of which 70 to 80% was soluble. Reactive lysine was 70 to 85%. Nutritional evaluation with rats indicated hydrolysates had 10 to 15% lower nutritional value than casein. Results suggest fish protein hydrolysates can be used in the food industry for their solubility and nutritional value.