Effect of dietary common carp by‐product protein hydrolysates on antioxidant status in different organs of zebrafish (Danio rerio)

A feeding trial was conducted to evaluate the effect of dietary protein hydrolysates from common carp (Cyprinus carpio) by‐products on the antioxidant status of zebrafish (Danio rerio). Common carp by‐product was hydrolysed using Alcalase to a degree of hydrolysis of 15%. The zebrafish were fed for 44 days with four different diets with increasing levels of carp by‐product hydrolysates (CBH0: 0 g/kg; CBH25: 25 g/kg; CBH50: 50 g/kg; CBH100: 100 g/kg). The gills, muscle and brain were dissected at the end of the feeding trial in order to evaluate the total antioxidant capacity against peroxyl radicals (ACAP) and lipid peroxidation (TBARS). Although total antioxidant capacity did not show differences in muscle (p > 0.05), lipid peroxidation was reduced in the muscle of fish fed the CBH50 diet. Brain lipid peroxidation showed a significant reduction (p < 0.05) in all groups when compared with the control diet CBH0. Antioxidant properties of protein hydrolysates indicate their potential as nutraceuticals since (a) a reduction in muscle lipid peroxidation was verified, implying that their use could enhance the quality and shelf life of fish fillets; and (b) a decrease in brain lipid peroxidation was registered, highlighting the potential use of fish protein hydrolysates for the prevention of neurodegenerative diseases.     

Optimization of Antioxidant Peptides Production from the Mantle of Cuttlefish (Sepia pharaonis) Using RSM and Fractionation

Persian Gulf cuttlefish mantles were hydrolyzed (CPH) using alcalase, and the optimal hydrolysis parameters were obtained for the highest degree of hydrolysis (DH) and strongest antioxidant (based on their ability to quench 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals) activity using response surface methodology (RSM). The predicted optimal parameters of DH and quenching DPPH radicals was: pH of 7.88, 50.2°C, 150 min, and enzyme to substrate ratio of 1.5%. The reducing power (RP) and ability of optimized peptides to quench ABTS radicals in a gastro-intestinal track model system increased during the intestinal stage, while scavenging ability against DPPH radicals dropped (P < 0.05). The oxidation of lipid was retarded in a lecithin-liposome model added with optimized CPH in a concentration dependent response. Ultrafiltration of optimized CPH showed that the 3–10 KDa fraction had the greatest DPPH radical scavenging activity, the 10–30 KDa fraction had the highest reducing power, and the <3 KDa fraction had the greatest ABTS radical scavenging activity.     

Common Kilka Hydrolysis: Investigation of Functional and In Vitro Antioxidant Properties of Hydrolysates Obtained by Kiwifruit and Ginger Proteases

The functional and in vitro antioxidant properties of common kilka fish protein hydrolysates with different degree of hydrolysis (DH) obtained by kiwifruit protease (KP) and ginger protease (GP) were evaluated. The electrophoretic patterns of hydrolysates showed the presence of one major band in different DHs with a molecular weight of less than 25 kDa. Protein solubility was positively correlated with DH, and the solubility of KP and GP hydrolysates (HKP and HGP, respectively) at 15% DH was higher than other DHs (p ≤ 0.05). Higher emulsifying and foaming properties were observed in HKP over a pH range of 2–10 (p ≤ 0.05), and in vitro antioxidant activity was higher in HKP at 15% DH as seen from 2,2-diphenyl-1-picrylhydrazyl radical scavenging and ferric-reducing power. Thus, the results reveal that DH and enzyme type affects functional properties and antioxidant activity of the hydrolysates.     

Utilization of Chum Salmon (Oncorhynchus keta) Skin Gelatin Hydrolysates to Attenuate Hydrogen Peroxide-Induced Oxidative Injury in Rat Hepatocyte BRL Cell Model

Six chum salmon skin gelatin hydrolysates with degree of hydrolysis of 4.7–13.5% were generated by Alcalase and papain and showed scavenging activities to 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion radicals. Potential protection of these hydrolysates against H₂O₂-induced oxidative injury in rat hepatocyte line (BRL cells) was assessed in vitro, based on cell viability, leakage of lactate dehydrogenase, thiobarbituric acid reactive substances (TBARS), glutathione content, glutathione reductase, and catalase of the cells. The hydrolysates exhibited protection on the hepatocytes, reflected by an enhanced cell viability (about 6.2–87.9%), glutathione reductase (about 28.2–85.9%) or catalase (about 43.3–116.9%) activity, and decreased lactate dehydrogenase leakage (about 2.7–34.7%) or TBARS content (about 4.1–39.3%). On the contrary, cellular glutathione content gave an unnoticeable difference among the investigated groups. These hydrolysates also showed protection against DNA damage in the cells. Cytoprotection of these hydrolysates possessed a dosage-dependence manner. More importantly, there was a high correlation (0.7 < r < 0.9) between DPPH or superoxide anion radical scavenging activity of these hydrolysates and three cellular antioxidant indices (glutathione content, glutathione reductase, and catalase activity).     

Assessing the Antioxidant Activity of the Ultrafiltration Fractions From Silver Carp Protein Hydrolysate by Different Antioxidant Methods

Silver carp protein hydrolysate (SH) was fractionated by 5- and 1-kDa cutoff ultrafiltration (UF) membrane, and three ultrafiltrates were obtained: SHI (>5 kDa), SHII (1–5 kDa), and SHIII (<1 kDa) which were screened for antioxidant activity by five in vitro assays. The SHIII showed higher 2, 2-azobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) values, inhibiting effects for liposome oxidation induced by Fe³⁺/V_C. The SHII showed higher ferric reducing power (FRAP) and inhibiting effects for liposome oxidation induced by 2, 2′-azobis (2-methylpropionamidine) dihydrochloride (AAPH). However, the SHI showed the highest 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging activity. Therefore, the antioxidant activity of silver carp protein hydrolysates was mainly associated with their molecular weight distribution, and the antioxidant assays used to test different samples gave comparable results.     

Antioxidant and Angiotensin I Converting Enzyme Inhibition Effects and Antihypertensive Effect in Spontaneously Hyertensive Rats of Peptide Isolated from Boiled Abalone By-Products,Hallotis discus hannai

ABSTRACT

Abalone is a marine gastropod that is an important fishery and food industrial resource, massively maricultured in Asia, Africa, Australia, and America. Marine by-products from boiled abalone, normally discarded as industrial waste in manufacturing plants, were fractionated and concentrated using an ultrafiltration membrane system (? 1 kDa). In this study, the antioxidant and angiotensin I converting enzyme (ACE) inhibition effects of the boiled abalone by-products (BABs) were investigated. BABs have high antioxidant activities against free radicals (1,1-diphenyl-2-pycryl-hydrazyl (DPPH), hydroxyl, peroxyl, and superoxide radicals), reactive oxygen species (ROS) stress, and DNA damage in H₂O₂-treated RAW 264.7 macrophages. The structure of the purified peptide was identified to be ATPGDEG (MW 752 Da), and the ACE inhibition pattern of the peptide was found to be noncompetitive. In addition, the peptide exhibited an antihypertensive effect according to time-course measurement after oral administration to spontaneously hypertensive rats (SHR). BABs from this genus may be potential candidates for the development of unique natural substances for further industrial applications as functional foods and pharmaceuticals.     

Antioxidant Properties of Scomber japonicus Hydrolysates Prepared by Enzymatic Hydrolysis

The antioxidant properties of the Pacific chub mackerel (Scomber japonicus) muscle protein hydrolysates prepared by enzymatic hydrolysis were investigated. After enzyme hydrolysis at 50°C for 60 min, more than 80% of the S. japonicus muscle protein was hydrolyzed. The highest 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity (71.69%) occurred in whole muscle protein hydrolysates treated at 50°C for 30 min with Protamex, and the highest 2,2?-azino-bis(3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) radical-scavenging activity (95.39%) was observed in white muscle protein hydrolysates treated at 50°C for 30 min with Neutrase. The highest superoxide dismutase (SOD)-like activity (32.84%) was recorded in white muscle protein hydrolysates treated at 50°C for 120 min with Protamex. Changes in the molecular weight distribution of S. japonicus muscle proteins after enzymatic hydrolysis were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A robust and a convenient enzyme hydrolysis technique for obtaining S. japonicus muscle protein hydrolysates with useful biological activities, within a short time (<2 h) is proposed.     

Enzymatic Hydrolysis of Catfish (Pangasius hypophthalmus) By-Product: Kinetic Analysis of Key Process Parameters and Characteristics of the Hydrolysates Obtained

In this study, kinetic evaluation of enzymatic hydrolysis of catfish (Pangasius hypophthalmus) by-product meat was conducted. The effects of key factors such as type of proteases (neutrase, papain, and bromelain), enzyme to substrate ratio (E/S), substrate concentration, and extent of hydrolysis on process efficiency, properties and functionalities of hydrolysates were evaluated. The results demonstrated that, for catfish shredded meat, bromelain and papain had higher affinity than neutrase. Kinetics parameters V_m and K_m of neutrase, papain, and bromelain were 4.486, 9.060, and 18.249 μM/min and 35.433, 6.582, and 7.034 g/l, respectively. Optimal E/S ratios of neutrase, papain, and bromelain were estimated to be 0.92, 2.76, and 3.31 mg/g. At optimal E/S, the highest possible substrate concentrations were found to be 108.4 g/l for neutrase, 36.2 g/l for papain, and 135.8 g/l for bromelain. Both kinetics study and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that bromelain and papain had higher proteolytic activity toward catfish protein than neutrase. A significant increase in antioxidant activity of hydrolysates was observed with neutrase and papain hydrolysate, but not with bromelain. The results could be helpful for industrial processors to develop an efficient process for catfish by-product valorization.     

Properties of Fish Sausages Containing Common Carp (Cyprinus carpio) Roe Oil and Defatted Roe Protein Hydrolysate during Refrigerated Storage

Influence of emulsified or non-emulsified common carp roe oil (CRO) and carp defatted roe hydrolysate (CDRH) on properties of crucian carp (Carassius auratus) sausages was determined during 30-day storage at 4°C. Sausages containing CRO and CDRH were lighter and had better textural properties including hardness, gumminess, and chewiness. These textural attributes were significantly higher in the sample prepared with pre-emulsified CRO and 7 g/100 g CDRH (p < 0.05). The sausages fortified with CDRH also presented better microstructure, as evidenced by fewer voids and smaller oil droplets. When incorporated at 7 g/100 g, CDRH could significantly reduce microbial spoilage in sausages in terms of the total viable count and the psychrophilic bacterial count (p < 0.05); however, pre-emulsification had no significant effect on antibacterial activity of hydrolysate in the samples (p > 0.05). Sausages with pre-emulsified roe oil and 7 g/100 g hydrolysate were more resistant against lipid oxidation and had a higher level of polyunsaturated fatty acids, especially docosahexaenoic acid and eicosapentaenoic acid after 30 days of storage at 4°C. Furthermore, the sausages with pre-emulsified CRO and 7 g/100 g CDRH exhibited better organoleptic properties at day 30.     

The effect of dietary folic acid on flesh quality and muscle antioxidant status referring to Nrf2 signalling pathway in young grass carp (Ctenopharyngodon idella)

To investigate the effect of dietary folic acid on fish flesh quality, muscle antioxidant status and the potential mechanism, young grass carp (Ctenopharyngodon idella) were fed diets containing 0.10–3.12 mg/kg diet of folic acid for 8 weeks. The results showed that optimum folic acid increased muscle contents of protein, lipid, pH, shear force, amino acids, unsaturated fatty acids, hydroxyproline and glutathione, whereas contents of moisture, cooking loss, lactate and saturated fatty acids and cathepsin L activity showed an opposite trend (p < .05). Moreover, optimum folic acid elevated antioxidant enzyme activities and mRNA levels, as well as NF‐E2‐related factor 2 and casein kinase 2 mRNA levels (p < .05). However, optimum folic acid decreased reactive oxygen species (ROS) content and Kelch‐like ECH‐associated protein 1a (Keap1a) and Keap1b mRNA levels (p < .05). Interestingly, excess folic acid induced negative effects on above‐mentioned parameters (p < .05). Summarily, this study indicated that optimum folic acid improved fish flesh quality and muscle antioxidant system associated with Nrf2‐Keap1 pathway. Based on muscle cathepsin L activity and ROS content, the folic acid requirements for young grass carp were 1.87 and 1.80 mg/kg diet, respectively, regarding to the flesh quality and antioxidant status.     

Evaluation of Plasticizing and Antioxidant Properties of Silver Carp Protein Hydrolysates in Fish Gelatin Film

Physico-mechanical and antioxidant properties of glycerol plasticized fish gelatin films were investigated at different levels of fish protein hydrolysates (FPH) from silver carp muscle (5, 10, 15, and 20 g FPH/100 g gelatin). The films supplemented with FPH, especially at higher concentration showed significant increase in 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 20-azinobis-(3-ethylbenzothiazoline-6-sulfnic acid) (ABTS) radical scavenging, and ferric reducing antioxidant power (FRAP), elongation at break, difference of color, water vapor permeability and opacity, while tensile strength, elastic modulus, L*, b*, and contact angle decreased markedly (p < 0.05). However, no significant differences were observed for a* and solubility of all samples (p > 0.05). Films prepared using 20% FPH had lower glass transition temperature and more homogeneous structure, compared with control film. Also, Fourier transform infrared (FTIR) spectroscopy confirmed the increase of free groups of hydrolysates and lower interaction between chains of film incorporated with the highest content of FPH. Thus, these results indicate that FPH enhanced the antioxidant activities and affected some characteristics due to less interaction between gelatin and FPH.     

Protein Hydrolysates from Shrimp (Metapenaeus dobsoni) Head Waste: Optimization of Extraction Conditions by Response Surface Methodology

Utilization of shrimp head waste from the processing industry offers a solution to environmental pollution and also produces valuable bioactive peptides. The present study was aimed to optimize the alcalase assisted extraction conditions by response surface methodology (RSM) to obtain protein hydrolysates from shrimp (Metapenaeus dobsoni) head waste with maximum antioxidant activity. Based on the models derived by RSM, the optimized conditions for maximum degree of hydrolysis (DH) were pH 8.2, temperature of 45.4°C, and enzyme/substrate (E/S) of 1.8%. Meanwhile, the optimized conditions for maximum antioxidant activity of protein hydrolysates as measured by 2, 2-diphenyl-1-picrylhydrozyl (DPPH) and ferric reducing antioxidant power (FRAP) assays were pH 8.2 and 8.1, temperatures of 49.7 and 59.4°C, and E/S of 2.1 and 2.7%, respectively. The maximum predicted values for DH, DPPH, and FRAP were 42.44%, 39.64% inhibition, and 8.3 μM Fe (II)/g wet wt. of sample, respectively. The extraction of protein hydrolysates at optimal conditions derived by these models resulted in DH, DPPH, and FRAPs of 40.31%, 38.93%, and 8.21 μM Fe (II)/g of sample, respectively, demonstrating the fitness of the models. The present study demonstrates the potential of Metapenaeus dobsoni head waste as a source of protein hydrolysates with prospective antioxidant activity.     

Application of Two-Level Full Factorial Design for the Extraction of Fucoxanthin and Antioxidant Activities from Sargassum siliquosum and Sargassum polycystum

Two-level full factorial design was employed to identify the extraction parameters that can improve the derivation of fucoxanthin content (FC), total carotenoid content (TCC), and antioxidant from two brown seaweeds, Sargassum siliquosum (SS) and Sargassum polycystum (SP). These parameters included temperature (A: 4–45°C), time (B: 30–1,440 min), and solvent-to-solid ratio (C: 10–50 ml/g). Antioxidant activities were determined as trolox equivalent antioxidant capacity (TEAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and ferric-reducing antioxidant power (FRAP). Results showed that all three factors were significant (p < 0.05) in providing higher FC in both species. These factors were also significant in obtaining higher TCC in SS; whereas in SP, TCC was only affected by solvent-to-solid ratio. Only temperature was found to contribute significantly to a higher TEAC in both species. However, none of the factors improved DPPH for SS, except temperature and time for SP. For SS, only time was significant in obtaining higher FRAP; whereas temperature and time were significant for SP. Hence, results indicate that a simple modification in the extraction temperature, time, and solvent-to-solid ratio will be able to improve the derivation of fucoxanthin, carotenoids, and antioxidant activities.     

Characterization of Collagen Enzymatic Hydrolysates Derived from Lizardfish (Synodus fuscus) Scales

Acid soluble collagen was extracted from the scales of lizardfish and characterized with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and amino acid analysis. After 8 h of collagen hydrolysis, hydrolysates had an estimated degree of hydrolysis (DH) from 4 ± 0.05 to 25 ± 0.63% (p < 0.05). All hydrolysates had angiotensin converting enzyme inhibitory and antioxidant activity. These activity levels showed little change after treatment with gastrointestinal proteases. Results indicated that the lizardfish by-products may be improved by enzymatic treatment with acid-soluble collagen from lizardfish scales.     

响应面法优化中性蛋白酶酶解海湾扇贝副产物制备抗氧化酶解物研究

采用响应面法优化中性蛋白酶酶解海湾扇贝(Argopecten irradias)副产物蛋白制备抗氧化酶解物工艺。以DPPH自由基清除率和还原能力为响应值,在底物浓度、温度、时间、酶与底物比4个单因素实验基础上,优化确定3个显著性因素。获得实验条件下最佳制备工艺为:底物浓度25 mg·mL^-1、温度50℃、时间3.8 h、酶与底物比1.5%。在此条件下,实验验证酶解海湾扇贝副产物蛋白制备抗氧化酶解物的DPPH清除率为81.93%(预测值为83.19%),还原能力为58.12%(预测值为58.92%),DPPH自由基清除率和还原能力的IC 50值分别为0.264 mg·mL^-1和3.136 mg·mL^-1,且表现出较强的抗氧化活性。研究表明,优化后的回归模型用于预测中性蛋白酶酶解海湾扇贝副产物蛋白制备抗氧化酶解物是可行的。     

Antioxidant and Angiotensin-Converting Enzyme Inhibitory Activities of Protein Hydrolysates Prepared from Threadfin Bream (Nemipterus spp.) Surimi By-products

Solid wastes from threadfin bream (Nemipterus spp.) surimi production composed of head and frame were hydrolyzed by various commercial proteases (Alcalase, Flavourzyme, Neutrase, Protamex, papain, and pepsin) to produce protein hydrolysates with bioactive properties. An Alcalase-hydrolyzed sample at 24.4% degree of hydrolysis (DH) displayed the highest antioxidant activity based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) assay, and potassium ferricyanide method. In addition, it showed an inhibitory activity toward angiotensin-converting enzyme (ACE) of 25.5%. Antioxidant activity of threadfin bream by-product hydrolysates increased with hydrolysis time and reached the highest DPPH activity after 6 h, while that hydrolyzed for 3 h showed the highest reducing power based on FRAP and potassium ferricyanide assays. In addition, ACE inhibitory activity was found to be at an optimum after 3 h of hydrolysis. The hydrolysates (1 mg/mL) also retarded oxidation of a linoleic acid emulsion system to a similar extent as 0.1 mg/mL 3-tert-butyl-4-hydroxy anisole (BHA), indicating a potential use in the food system. Protein hydrolysates from threadfin bream surimi by-products could be tailor-made to possess both antioxidant and ACE inhibitory activity through controlling DH of Alcalase-catalyzed reactions.     

Anti-Inflammatory,Antioxidant, and Antimicrobial Effects of Underutilized Fish Protein Hydrolysate

Antimicrobial, anti-inflammatory, and antioxidant activities of protein hydrolysates from Argentine croaker (Umbrina canosai) protein isolate (CPI) or Argentine croaker myofibrillar protein with different degrees of hydrolysis (DH: 10–20%) prepared using Alcalase or Protamex were determined. Results showed that an increase in the DH resulted in higher content of hydrophobic and aromatic amino acids (AAAs) and in a decrease in molecular weight (MW) distribution for all hydrolysates obtained. Furthermore, the enzyme and raw material used influenced the amino acid content and MW determined. Hydrolysates from CPI with a 20% DH by Alcalase had higher 2,2?-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity, metal chelation, and ferric-reducing antioxidant power (p < 0.05). All hydrolysate samples decreased the pro-inflammatory capacity. In all the evaluated microorganisms, only seven were inhibited, most being Gram-positive. Alcalase was found to exert a considerable influence on antibacterial activity. These hydrolysates are an alternative as natural antimicrobials, anti-inflammatory, and antioxidant compounds.     

Stimulatory effect of dietary taurine on growth performance,digestive enzymes activity,antioxidant capacity,and tolerance of common carp, <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L., fry to salinity stress

The present study was carried out to evaluate the effect of dietary taurine (Tau) on performance, digestive enzymes, antioxidant activity, and resistance of common carp, Cyprinus carpio L., fry to salinity stress. Fish (0.97?±?0.033 g) were fed on different taurine levels of 0.0 (control), 5, 10, 15, or 20 g/kg diet up to satiation twice daily for 8 weeks. At the end of the feeding trial, fish were stressed by exposure to 10 ppt salinity for 3 days during which fish mortality was observed. Fish performance was significantly (P?<?0.05) improved by dietary taurine up to 15 g Tau/kg diet after which fish growth and feed intake were almost the same. Also, taurine supplementation significantly (P?<?0.05) elevated activities of intestinal amylase, lipase, and protease resulting in an improving in feed intake giving better performance. Furthermore, Tau-stimulated antioxidant activity of common carp was observed in a dose-related manner, where activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly (P?<?0.05) higher, but malondialdehyde (MDA) value was significantly (P?<?0.05) lower in Tau-fed fish groups than those fed the control diet. In salinity stress experiment, highest survival rate was observed at fish fed Tau-supplemented diets without significant (P?>?0.05) differences over fish fed the control diet. It appears that taurine could be used as a feed supplement to confer better growth and health of common carp fry with optimal level of 15 g/kg diet.     

Assessment of the Nutritive,Biochemical, Antioxidant and Antibacterial Potential of Eight Tropical Macro algae Along Kachchh Coast,India as Human Food Supplements

Eight tropical marine macro algae were investigated for in-vitro antioxidant, antibacterial, and biochemical properties. The moisture content [% DW (dry weight)] ranged from 7.21 to 14.72%; ash, 24.92 to 47.04%; lipids, 0.73 to 2.67%; protein, 4.56 to 12.59%; and carbohydrate, 30.1 to 48.51%. The % sum of polyunsaturated fatty acids (PUFAs) ranged from 21.71 to 78.22%, while omega-6/omega-3 (ω6/ω3) ratio was from 0 to 2.08, which remained within the prescribed World Health Organization (WHO) standards (<10). The % sum of essential amino acids (EAAs) ranged from 86.18 to 204.66, and Na/K ratio ranged from 0.37 to 2.85. The extracts exhibited significant (P < 0.05) values for total phenol (7.25–16.0 µg PGE mg^–1 DW) and total flavonoid (4.06–15.63 µg QE mg^–1 DW) at 200 µg/ml. Antibacterial activity was tested against the selected food spoilage and pathogenic bacteria. The principal component analysis (PCA) confirmed a positive correlation between total phenolic content, 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging activities, and ferric reducing antioxidant power (FRAP), which supports the integration of seaweeds as ingredients in functional foods.     

Improvement of Antioxidant Activity of Grass Carp (Ctenopharyngodon idella) Protein Hydrolysate by Washing and Membrane Removal Pretreatments and Ultrasonic Treatment

To improve the antioxidant activity of grass carp hydrolysate, washing and membrane removal pretreatments and ultrasonic treatment were applied in this study. Various pretreatment methods (washing methods and membrane removal methods) for removing the prooxidants and phospholipid in minced fish were evaluated. The effects of ultrasonic treatment prior to enzyme hydrolysis on the antioxidant activities of minced carp hydrolysate were investigated. The minced carp subjected to washing with 0.4% NaCl solution and subsequent membrane removal with CaCl₂ and citric acid solution had the lowest content of prooxidants and phospholipid remaining in the minced fish. The hydrolysate produced from washing/membrane removal pretreated carp with ultrasonic treatment for 20 min had the highest ferric reducing antioxidant power and scavenging activities against 1,1-diphenyl-2-picrylhydrazyl and 2,2?-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radicals. This suggested that the combination approach of washing/membrane removal pretreatment and ultrasonic treatment could improve the antioxidant activity of carp hydrolysate, and this hydrolysate could be a potential antioxidant ingredient for functional foods.