Potential Precursor of Angiotensin-I Converting Enzyme (ACE) Inhibitory Activity and Structural Properties of Peptide from Peptic Hydrolysate of Cutlassfish Muscle

ABSTRACT

Peptic hydrolysates were prepared by digesting the cutlassfish muscle protein using pepsin for 1, 3, and 6 h, and their inhibitory activity against angiotensin-I converting enzyme (ACE) was studied. The ACE-inhibitory effect of the peptic hydrolysate of cutlassfish muscle generated at the 3 h time point exhibited the strongest activity. After identifying the optimal hydrolysate, the active peptide was isolated by ultrafiltration, gel permeation, and high performance liquid chromatography (HPLC). The resulting purified peptide was characterized using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/TOF MS/MS) and was identified to be a 496.44 Da pentapeptide (Phe-Ser-Gly-Gly-Glu). The ACE-inhibitory activity of the active peptide exhibited an IC₅₀ value of 0.033 ± 0.003 mg/ml. A molecular docking program was used to simulate the interaction between the peptide and ACE, which revealed that the inhibitory effect was mainly due to the hydrogen bonds between ACE and the peptide. Based on the ACE-inhibitory properties and the molecular docking study of the resulting active peptide, we demonstrated an increase in nitric oxide (NO) production in a dose-dependent manner. In conclusion, cutlassfish protein hydrolysate and the resulting active peptide could be used as active ingredients in functional food as anti-hypertensive agents.     

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.     

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.     

Inhibitory Activities of Protein Hydrolysates from Spotted Babylon Snails on Tyrosinase and Melanogenesis

The optimal conditions for protein hydrolysates preparation from spotted babylon Babylonia areolata exhibiting tyrosinase inhibitory activity and antioxidant activity using alkaline protease (Protease G6) were undiluted Protease G6 (5.8 × 10⁵ DU/g) at 60 min of hydrolysis time and eightfold diluted Protease G6 (7.25 × 10⁴ DU/g) at 240 min of hydrolysis time. These two conditions were fractioned using molecular weight (MW) cutoff values of 10, 5, and 3 kDa membranes, and their anti-melanogenic and antioxidant properties were further analyzed. Among the fractions, the MW < 3 kDa fraction exhibited high levels of inhibitory activity toward the mono- and diphenolase activities of tyrosinase, with IC₅₀ values of 1.758 and 8.995 μg/mL, respectively. Kinetic studies revealed that this fraction behaved as an uncompetitive inhibitor. The results demonstrated that the MW < 3 kDa fraction suppressed melanin synthesis and decreased cellular tyrosinase activity with no cytotoxicity to B16F10 melanoma cells.     

Purification and Antioxidant Property of Antioxidative Oligopeptide from Short-Necked Clam (Ruditapes philippinarum) Hydrolysate In Vitro

Short-necked clam (Ruditapes philippinarum) muscle was hydrolyzed with trypsin, and the antioxidant activities of isolated peptides were investigated. The hydrolysate was fractionated by chromatographic methods using Sephadex G-25 gel filtration and Hypersil BDS C18 reversed phase high performance liquid chromatography (RP-HPLC). The antioxidant activities of the hydrolysate were determined by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH), reducing power assay, and the protective effect against hydroxyl-radical-induced DNA damage. The amino acid sequence of the purified antioxidant oligopeptide was found to be Gly-Asp-Gln-Gln-Lys.     

Protein Hydrolysate from Visceral Waste Proteins of Bolti Fish (Tilapia nilotica): Chemical and Nutritional Variations as Affected by Processing pHs and Time of Hydrolysis

The main objective of the current work was to evaluate the proximate chemical composition and nutritional quality of amino acids of protein hydrolysates from viscera of Tilapia nilotica fish. The results of amino acid composition showed that all hydrolysates contained sufficient and/or excessive amounts of both essential and nonessential amino acids, with appreciable values of essential amino acid/nonessential amino acid ratio (0.77 to 1.59) and essential amino acid/total amino acid ratio (43.35 to 61.35). All protein hydrolysates had a high content of the flavor enhancer amino acids. The lowest values of essential amino acids were found for phenylalanine and methionine with chemical scores ranging from 0.04 to 0.77 and 0.09 to 1.06, respectively, except for 18 h hydrolysates. The protein efficiency ratio had values greatly exceeding 2.0, which is considered a high quality. On the other hand, the levels of sugar and ash contents ranged from 2.69 to 3.32 g L^?1 and 0.48 to 2.1%, respectively, with 9.2 to 14% dry matter. Mineral composition showed that sufficient amounts of calcium, sodium, potassium, magnesium, and iron were present. The results suggest that the studied hydrolysates are appropriate for use in balanced human or animal diets.     

Purification and Identification of Antioxidant Peptides from Fermented Fish Sauce (Budu)

This study aimed to isolate antioxidant peptides from Budu extract and identify novel antioxidant peptides using high-performance liquid chromatography (HPLC) and electrospray ionization-time-of-flight mass spectrometry (ESI-TOF MS/MS). An amino acid analyzer was used to analyze the amino acid composition of peptides. Two novel peptides, Lue-Asp-Asp-Pro-Val-Phe-Ile-His (LDDPVFIH) and Val-Ala-Ala-Gly-Arg-Thr-Asp-Ala-Gly-Val-His (VAAGRTDAGVH), were identified. The synthesized peptide of LDDPVFIH showed higher antioxidant activity. The presence of hydrophobic amino acids (Ile and Leu), acidic (Asp) and basic (His) amino acids in the peptide sequences is believed to contribute to the high antioxidant activity of the fermented anchovy fish (Budu) extract. Thus, the two peptides may have potential application as functional foods and could also be used as nutraceutical compounds.     

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

Trypsin 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.     

Effect of Enzyme Type on the Antioxidant Activities and Functional Properties of Enzymatic Hydrolysates from Sea Cucumber (Cucumaria frondosa) Viscera

ABSTRACT

Effects of various proteases were obvious on the amino acid composition, antioxidant activities, and functional properties of enzymatic hydrolysates prepared from sea cucumber viscera (SCV). Alcalase, trypsin, and flavourzyme conferred better antioxidant activities on hydrolysates and endowed them with a higher degree of hydrolysis, coming to 19.08, 32.38, and 15.94%, respectively. For functional properties, neutrase- and flavourzyme-derived hydrolysates showed better foaming properties. Amino acid composition suggested that papain, neutrase, alcalase, and bromelain granted the hydrolysate’s high nutritional quality. The results indicated that SCV hydrolysates could be used in food systems to impart desirable characteristics to products.     

大眼金枪鱼头蛋白酶解物1 ku超滤组分的抗氧化活性及其理化性质

研究大眼金枪鱼头蛋白酶解物1 ku超滤组分体外的还原力、自由基清除能力及对衰老小鼠体内抗氧化能力的影响,分析1 ku超滤组分的一般成分、氨基酸组成及分子量分布,为进一步分离纯化金枪鱼头抗氧化肽提供基础。体外结果显示,1 ku超滤组分对羟基自由基、超氧阴离子和DPPH自由基的清除活性随浓度的增加而增强,IC50分别为1.38、0.73与0.93mg/mL,还原力也随浓度的增加而增大,在浓度为12.5 mg/mL时为0.763;体内结果显示,灌胃30 mg/kg的1 ku超滤组分连续42 d,D-半乳糖致衰老小鼠肝组织的超氧化物歧化酶(SOD)活性、肝组织和血清的谷胱甘肽过氧化物酶(GSH-PX)活性显著提高(P<0.05),血清丙二醛(MDA)含量显著降低(P<0.01);理化分析结果显示,1 ku超滤组分(干基计)蛋白质含量为96.40%,脂肪0.11%,灰分4.86%,疏水性氨基酸占氨基酸总量的35.8%,活性组分分子量在1 802~2 519 u和422~922 u。     

Golden Grey Mullet (Liza aurata) Alkaline Proteases: Biochemical Characterization,Application in the Shrimp Wastes Deproteinization,Laundry Commercial Detergents,and Preparation of Antioxidant Protein Hydrolysate

Digestive alkaline proteinases from golden grey mullet (Liza aurata) were extracted and characterized. The crude alkaline protease showed optimum activity at pH 8.0 and 60°C, and it was highly stable over a wide range of pH from 4.0 to 10.0, retaining more than 80% activity after incubation for 1 h at 4°C. The alkaline proteases showed extreme stability toward nonionic and anionic surfactants after preincubation for 1 h at 25°C and relative stability toward oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various solid and liquid detergents. Further, proteases from golden grey mullet viscera were found to be effective in the deproteinization of shrimp wastes. The protein removal after 3 h at 45°C with an enzyme/substrate (E/S) ratio of 10 U/mg protein was about 76%. The golden grey mullet proteases were also shown to be efficient in the production of antioxidant protein hydrolysate.     

四种加工方式对皱纹盘鲍制品消化特性的影响

为了探讨不同加工方式对皱纹盘鲍制品消化特性的影响,分析了生鲜、煮制、罐制和干制4种方式对鲍鱼肌肉蛋白在消化酶作用下的分解情况。利用模拟胃肠液消化及SDS-PAGE比较分析不同加工方式对鲍鱼肌肉消化特性的影响,并通过扫描电镜观察4种加工方式下鲍鱼肌肉组织微观结构的差异。结果显示,与生鲜鲍鱼相比,经煮制的鲍鱼较易被消化,罐制加工产品最易被消化,而干制鲍鱼最难被消化。不同加工方式对鲍鱼肌肉的组织结构有较大影响。进一步对不同皱纹盘鲍制品模拟胃肠液消化终产物的生物活性进行研究,发现它们对血管紧张素转换酶(ACE)的抑制活性(IC50)依次为罐制(464.2μg/m L)干制(665.4μg/m L)煮制(775.7μg/m L)生鲜(803.9μg/m L)。研究表明,不同加工方式对鲍鱼制品在机体内的消化特性及产物对ACE抑制活性存在差异,为优化鲍鱼制品生产加工提供了一定的参考。     

低鱼粉饲料中添加低分子水解鱼蛋白对凡纳滨对虾非特异性免疫力和抗氧化能力的影响

以鱼粉水平为30%的饲料为高鱼粉饲料对照组,以豆粕替代高鱼粉饲料中50%鱼粉形成低鱼粉基础饲料,低鱼粉基础饲料添加0、0.5%、1.0%、1.5%和2.0%低分子水解鱼蛋白,配制 6种实验饲料,对初始体重为(0.44±0.02) g的凡纳滨对虾(Litopenaeus vannamei)幼虾进行48 d的养殖实验。实验结束后采集样品,测定肝胰腺酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、总超氧化物歧化酶(T-SOD)活力和丙二醛(MDA)含量,并分析肠道先天免疫缺陷基因(IMD)、对虾素3a分子(Penaiedin 3a)、溶菌酶(LZM)和Crustin相关免疫基因的表达量,以探讨低分子水解鱼蛋白对凡纳滨对虾非特异性免疫力和抗氧化能力的影响。结果显示,在低鱼粉饲料中添加适宜水平的低分子水解鱼蛋白能够显著提高对虾肝胰腺T-SOD、AKP和ACP的活力,降低MDA的含量;且1.5%低分子水解鱼蛋白能增加IMD、Penaiedin 3a、LZM和Crustin基因表达量。综上,凡纳滨对虾非特异免疫力和抗氧化能力的降低可能是影响植物蛋白源替代鱼粉的关键原因之一,而在饲料中补充1.0%–1.5%的低分子水解鱼蛋白可有效提高凡纳滨对虾抗病相关基因的表达量,增强其非特异免疫力和抗氧化能力。     

不同蛋白水解物对花鲈(Lateolabrax japonicus)生长性能及非特异性免疫的影响

实验以基础饲料组[含30％鱼粉蛋白(FM)]为对照组,用鱼蛋白水解物(FPH)、猪血蛋白水解物(PBPH)、酵母蛋白水解物(YPH)和豆粕蛋白水解物(SPH)分别替代配方中10％的鱼粉蛋白,配制成5组等氮等脂的饲料,喂养初始平均体重为31.99 g的花鲈(Lateolabrax japonicus)8周,探讨饲料中不同蛋白水解物对花鲈生长、饲料利用、体组成成分及非特异性免疫的影响.结果显示:各处理组实验鱼存活率在97.78％-98.89％之间,没有显著性差异(P＞0.05);FPH组与FM组的末重和特定生长率最高,两者之间没有显著差异(P＞0.05),但显著高于PBPH组、SPH组和YPH组(P＜0.05);FPH组饲料效率高于FM组、PBPH组和YPH组(P＞0.05),且显著高于SPH组(P＜0.05);各实验处理组鲈鱼摄食率没有显著差异(P＞0.05);FPH组蛋白效率比显著高于FM组、PBPH组、SPH组和YPH组(P＜0.05);FPH组、FM组和YPH组的蛋白质沉积率显著高于PBPH组和SPH组(P＜0.05).饲料中添加FPH和YPH会显著增加花鲈肝脏和血清中碱性磷酸酶、酸性磷酸酶、超氧化物歧化酶、溶菌酶活性和总抗氧化能力,显著高于FM组、PBPH组和SPH组(P＜0.05).FPH组的花鲈鱼体粗蛋白含量高于FM组和YPH组,但差异不显著(P＞0.05),FPH组显著高于PBPH组和SPH组(P＜0.05).综上所述,4种不同蛋白水解物替代鱼粉后投喂花鲈幼鱼,鱼蛋白水解物效果最好,其次是酵母蛋白水解物、猪血蛋白水解物和豆粕蛋白水解物.     

虾头自溶产物中ACE抑制肽的分离鉴定

虾头在一定的条件下发生自溶作用,其所含蛋白质以肽和氨基酸等形式释放出来,有些肽产物具有ACE抑制活性.实验采用8000、5000和3000 u的超滤膜分级分离虾头自溶产物,活性检测结果表明,ACE抑制肽主要分布在3000 u超滤组分中;3000 u超滤组分进一步经Sephadex G-25葡聚糖凝胶层析、SP Sephadex C-25离子交换层析及Sephadex G-15葡聚糖凝胶层析纯化,ACE抑制活性提高将近8倍(IC50 =0.19 mg/mL);Sephadex G-15葡聚糖凝胶层析收集的高活性成分再经两次RP-HPLC纯化,分离纯化得到两条ACE抑制肽,质谱分析推测其氨基酸序列为Tyr-Pro和Leu-Pro/Ile-Pro,分子量分别为279和229 u.     

Evaluation of the Antioxidant Activity In Vitro and in Hippocampal HT-22 Cells System of Protein Hydrolysates of Common Carp (Cyprinus carpio) By-Product

Fish processing by-products may become more than 50% of the starting material. If mismanaged, these large quantities of discarded fish can create serious pollution problems and can also generate cost associated with their disposal. Enzymatic hydrolysis is one of the techniques that is currently being developed in order to recover and add value to these biomolecules. There is an increasing interest in natural antioxidants that are safer for consumers compared with synthetic antioxidants. In this study, common carp by-product was hydrolyzed using the enzymes Alcalase (A) and Protamex (P) to reach degrees of hydrolysis (DH) of 10 and 15%, respectively. Antioxidant capacity against peroxyl radicals (ACAP); 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging; and the measurement of intracellular reactive oxygen species (ROS) concentration after exposure to common carp protein hydrolysates were investigated. The results revealed that the hydrolysate A15 exhibited significantly (p < 0.05) higher antioxidant activity against the DPPH radical. A15 showed the highest in vitro antioxidant competence against peroxyl radicals, whereas P15 showed the lowest activity against peroxyl radicals (p < 0.05). Hydrolysates having the highest and the lowest in vitro antioxidant activity (A15 and P15, respectively) were selected for the determination of antioxidant activity in the HT-22 cells system. Measurement of intracellular ROS concentration revealed that P15 at the concentration of 1.25 mg/mL significantly (p < 0.05) reduced the intracellular ROS concentration. These results showed that common carp by-product protein hydrolysates are a source of antioxidant peptides with a high potential for food and pharmaceutical industries to develop new nutraceuticals and functional foods.     

Angiotensin Converting Enzyme and Dipeptidyl Peptidase-IV Inhibitory,and Antioxidant Activities of a Blue Mussel (Mytilus edulis) Meat Protein Extract and Its Hydrolysates

ABSTRACT

Protein extracted from mussel processing coproducts was hydrolyzed with four different food-grade enzyme preparations and assessed for angiotensin converting enzyme (ACE) and dipeptidyl peptidase-IV (DPP-IV) inhibitory and oxygen radical antioxidant capacity (ORAC) activities. All hydrolysates tested showed higher activity than the intact protein. ACE and DPP-IV IC₅₀ values in the range 1.13–3.34 and 0.33–2.43 mg mL^?1, respectively, and ORAC values in the range 66.26–121.56 µmol trolox equivalents g^–1 were obtained. These results suggest that some of the mussel meat protein hydrolysates may have potential as functional food ingredients for the management of diseases such as type II diabetes and hypertension.     

Purification and Characterization of an Antioxidant Protein from Pearl Oyster (Pinctada fucata martensii)

Protein from pearl oyster (Pinctada fucata martensii) was purified and characterized. Results showed that the pearl muscle protein (PMP) was composed of a single polypeptide with a molecular weight of approximate 11.6 kDa and contained 95.1% of the protein and 4.92% of the carbohydrate of the oyster. The polypeptide appears to be a glycoprotein, since the Fourier transform infrared (FTIR) spectroscopy of the PMP shows the typical characteristics of protein and polysaccharides. The denaturation temperature of the PMP was 81.3°C by differential scanning calorimetry. Circular dichroism spectroscopy showed that the PMP has a highly ordered structure. The PMP (200 μg/mL) showed a high oxygen radical absorbance activity (ORAC) of 6.57 μmol Trolox/μmol protein. Its antioxidant activity was stable at temperatures from 30–80°C and pH 2–10. The antioxidant activity was significantly inhibited by Zn²⁺, Ca²⁺, and EDTA and enhanced by Mn²⁺. The results suggest that the PMP could be used as a potential natural antioxidant.     

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.     

Influence of Degree of Hydrolysis on Chemical Composition,Functional Properties,and Antioxidant Activities of Chinese Sturgeon (Acipenser sinensis) Hydrolysates Obtained by Using Alcalase 2.4L

Protein hydrolysates from Chinese sturgeon were prepared using Alcalase 2.4L enzyme. Under the optimum conditions (enzyme–substrate ratio of 3.5%, pH of 8.5, and temperature of 55°C), the degree of hydrolysis (DH) was 13.8%, 16.7%, and 19.1% after 1, 3, and 6 h, respectively. The contents of crude protein and amino acid increased at DH of 19.1% to 86.97% and 78.29%, respectively. There was an obvious increase in the low-molecular-weight peptides, which could enhance the hydrolysate’s functional properties such as solubility, representing more than 90% at different pH levels. The obtained protein hydrolysates revealed good emulsification properties and high oil absorption. Furthermore, good antioxidant activities such as 1, 1-diphenyl-2-picrylhydrazyl (DPPH) inhibition, ferric reducing power, and ferrous ion (Fe²⁺⁾ chelating ability were attained depending on the solution concentration. The findings indicate that the functional and antioxidant properties of protein hydrolysates could be useful in many applications of the food industry.