Anchovy Protein Research Articles

Elaboration of cookies with pineapple peel flour from passion fruit (Passiflora edulis) enriched with anchovy protein concentrate (Engraulis ringens)

Los residuos de frutas tienen múltiples usos en la industria alimentaria tal como lo es en la elaboración de harina de cascara de maracuyá. A su vez, se observó que este alimento ofrece un aporte nutricional valioso para el organismo. En los últimos tiempos, se ha producido un avance notorio en la industria alimentaria, con un enfoque creciente en la creación de productos innovadores y sostenibles. En ese contexto, la investigación se centró en la preparación de galletas donde se utilizan ingredientes no convencionales, como el maracuyá, que se caracterizan por su riqueza en compuestos bioactivos y antioxidantes, así mismo se encontró su formulación óptima usando el 95 % de harina de trigo, 3 % de harina de cascara y un 2 % de concentrado proteico hidrolizado de anchoveta. Un aspecto particularmente interesante del estudio fue la incorporación de concentrado proteico hidrolizado de anchoveta, una fuente rica en proteínas y omega-3. Además, se realizó un análisis físico sensorial resultando en una aceptación satisfactoria, se observó que estas galletas ofrecen un aporte significativo de nutrientes beneficiosos para la salud.

Development of patty meat analogue using anchovy protein isolate (Stolephorus insularis) as a binding agent

The development of meat analogues focuses on sustainable production and requires attention to their nutritional, physicochemical, and sensory values. Anchovy protein isolate (API) is a novel and potential binding agent in the development of meat analogues. This study aimed to produce API and evaluate the physical, proximate, and sensory qualities of patty meat analogue (PMA) with the addition of API. The preparation method for API uses pH-shifting. The ratios of API added to the meat analogues were 0 % (F0), 4 % (F1), 8 % (F2), and 12 % (F3) per textured vegetable protein (TVP) weight. Furthermore, PMA was analysed for physical, proximate, and sensory properties. API had 87.23 % dry basis (db) protein content. The amino acid composition of API generally complied with the nutritional requirements of adults and children. The addition of API significantly affected the physical properties, proximate composition, and sensory (taste) qualities of PMA (p < 0.05). The protein content of PMA met Indonesian national standards (SNI) and was similar to both McDonald's and ground beef patty based on United States Department of Agriculture (USDA) standards. F3 was found to be the best based on its physical, proximate, and sensory properties.

Anchovy’s protein as a potential precursor of Angiotensin-I Converting Enzyme (ACE) inhibitory peptide and Dipeptidyl Peptidase-IV (DPP-IV) inhibitory peptide by an in silico approach

Protein from fish is known as the precursor of biologically active peptides that exert various health benefits such as antihypertensive, antitumor, and immunomodulatory properties. Hence, this study aimed to perform an extraction of anchovy, LC-MS/MS analysis and in silico evaluation of the major proteins in anchovies as potential precursors of biologically active peptides in addition to determining whether such peptides can be released by selected proteolytic enzymes. Anchovy was subjected to protein extraction followed by total soluble protein concentration determination using Bradford assay. The sample was subjected to in-solution trypsin digestion, which was then analysed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). A bioinformatic approach by PEAKS Studio was used to identify the protein. A total of four anchovy’s proteins which are myosin light chain 1, V(D)J recombination-activating protein, 60 kDa chaperonin and heat shock protein 90AA1 were identified. Then, the identified proteins were subjected to in silico approach using the BIOPEP database. The biological potential of the theoretically released angiotensin-I converting enzyme (ACE) inhibitory peptides and dipeptidyl peptidase (DPP)-IV inhibitory peptides were predicted by determining the frequency of occurrence of fragments with a given activity. 60 kDa chaperonin and heat shock protein 90AA1 predicted the highest number of biological activities for ACE inhibitory peptides (284 and 264 fragments) and DPP-IV inhibitory peptides (395 and 409 fragments). The most promising enzyme for the generation of bioactive peptides from anchovy protein was anticipated to be pepsin (pH &gt; 2), which theoretically released a high number of DPP-IV inhibitory peptides and ACE inhibitory peptides through the action of in silico proteolysis. Overall, this work highlighted that anchovy protein could be a promising precursor of bioactive peptides that have ACE and DPP-IV inhibitory activities for developing functional food or nutraceutical products.

Heterologous expression and characterization of Bacillus velezensis SW5 serine protease involved in the hydrolysis of anchovy protein.

Bacillus velezensis SW5, with good enzyme production ability, was isolated and identified in our laboratory from fermented fish sauce. Its galactosidase has been expressed in Escherichia coli, which could hydrolyze lactose in milk. The present study aims to express a novel serine protease gene (SPr-SW5) of this strain by Bacillus subtilis WB800N, and applies the expressed enzyme in hydrolysis of anchovy to prepare antioxidant substances, aiming to alleviate the waste of low-value fish resources. SPr-SW5 with the open reading frame of 1353 bp encodes a serine protease (SPr-SW5) with 450 amino acids. The theoretical molecular weight and isoelectric point are 47.2kDa and 5.22, respectively. The successful expression of SPr-SW5 in B. subtilis WB800N was confirmed by a skim milk plate test. Its optimal temperature and pH were 50 °C and 8.0, respectively. SPr-SW5 activity was increased by Ca2+ and Zn2+ , but inhibited by Fe3+ . Furthermore, SPr-SW5 was tolerant to 1% Tween-40 and Tween-80; however, its activity was strongly inhibited by 10mm phenylmethylsulfonyl fluoride. Additionally, SPr-SW5 could be capable of hydrolyzing anchovy, the hydrolysate (AHP) at 10g L-1 , with 2,2-diphenyl-1-picrylhydrazyl and hydroxyl (·OH) scavenging rates of 73.21% and 79.71%, displaying good antioxidant activity. The novel SPr-SW5 was successfully expressed in B. subtilis WB800N. It exhibited excellent temperature stability and good tolerance to several metal ions. In addition, the anchovy hydrolyzed by expressed SPr-SW5 has good antioxidant ability. Overall, this research lays a good foundation for SPr-SW5 with respect to exploration and application in the food industry as enzyme preparation. © 2023 Society of Chemical Industry.

Physicochemical and Sensory Properties of Protein Isolate from Anchovy (Stolephorus insularis)

Anchovy is one of the primary fishery commodities in Indonesia; however its development as fishery products is currently suboptimal. Due to its high protein content, anchovy is potential to be developed as a source of value-added fish protein isolate. This study aims to produce anchovy protein isolate (API) and evaluate its physical, chemical, and sensory properties. The API was prepared from defatted anchovy flour. Isolation of anchovy protein was carried out using a pH-shifting method. The API was then analyzed for its physicochemical (bulk density, color, proximate) and sensory properties. The results showed that anchovy protein was more soluble at pH 11 and less at pH 5. Yield and protein recovery of API were 26.39 and 36.86% wb, respectively. The API had 92.20% protein, 3.64% moisture, 2.18% ash, 2.26% lipid, and 3.36% carbohydrate content on a dry basis. The results showed that the API exhibited good physical and sensory properties such as bulk density, color, the best score on sweetness, seaweed, bitterness, off-flavor, aroma, and rancid taste. There was no significant difference in sweet taste, off-flavor, aroma, and rancid taste between API and soy protein isolate (SPI) (p0.05). Overall, API demonstrated satisfactory nutritional properties and potential use as food ingredients.

Evaluation of taste active peptides and amino acids from anchovy proteins in fish sauce by insilico approach.

Enzymatic activity and microbial fermentation play a prominent role in the bioconversion of complex muscle tissue into smaller units of peptides and amino acids, possibly contribute to sensory properties. Thus, this study screens and evaluate anchovy proteins with taste-active peptides and amino acids by the reaction of multiple enzymes using an insilico approach. Information about sensory components was provided based on an insilico analysis using tools available in the BIOPEP-UWM database. Proteins from anchovy, namely myosin heavy chain 6 alpha, myosin light chain 1, cytochrome B, and NADH ubiquinone oxidoreductase, were subjected to insilico digestion with the combination of 23 enzymes. This led to the release of taste-active peptides and amino acids, including umami, sweet, salty, sour, and bitter sensory properties. The combination of multiple enzymes released a more significant number of taste-active peptides and amino acids for both myosins compared to other proteins.

Fabrication and characterization of anchovy protein hydrolysates-polyphenol conjugates with stabilizing effects on fish oil emulsion

Conjugation between peptides and polyphenols could improve their bioactive and functional properties. The improvement effects of anchovy protein hydrolysates (APH) -polyphenol (catechin (CA), gallic acid (GA), tannic acid (TA)) conjugates were investigated. The content of protein and polyphenols and ratio of polyphenols/peptides in conjugates increased as the number of OH group increased with TA > CA > GA. Results showed that APH-CA and APH-GA exhibited the highest ORAC and ABTS+ scavenging capacity, respectively. Mass spectrometry analysis suggested the highest number of bioactive peptides were identified in APH-CA 5:1 (APH/polyphenols). The physical stability of fish oil emulsions during storage was significantly enhanced by TA 5:1 conjugate followed by CA 5:1 conjugate. The oxidative stability was remarkably elevated by APH-GA 10:1. This was due to the antioxidant capacity and the peptides adsorbed at the interfacial. This study demonstrated that APH-polyphenol conjugates could bring the possibility of utilizing peptides-polyphenols in the nutraceutical and functional food ingredient fields.

Administration of Protein Hydrolysates from Anchovy (Engraulis Encrasicolus) Waste for Twelve Weeks Decreases Metabolic Dysfunction-Associated Fatty Liver Disease Severity in ApoE-/-Mice.

Simple SummaryMetabolic dysfunction-associated fatty liver disease (MAFLD) is an increasing concern worldwide. It currently represents the main cause of chronic liver disease in humans in Western countries. Nutritional strategies based on fish-rich diets are considered helpful in the prevention of MAFLD, and are also thought to be beneficial for human health. In particular, cholesterol- and triacylglycerol-lowering effects are associated with fish-derived proteins or hydrolysates. Our findings suggest that supplementing the diet with 10% (w/w) anchovy protein hydrolysates has an anti-obesity effect together with an improvement in lipid metabolism and a reduction in liver fat content and high-fat diet-induced liver disease. By virtue of their nutritional value and functional proprieties, anchovy by-product protein hydrolysates may be an efficient nutritional strategy in MAFLD prevention and treatment.Metabolic dysfunction-associated fatty liver disease (MAFLD) includes several diseases, ranging from simple steatosis to steatohepatitis, fibrosis and cirrhosis. Fish-rich diets are considered helpful in the prevention of MAFLD, and the enzymatic hydrolysis of fish waste has been explored as a means of obtaining high-value protein hydrolysates, which have been proven to exert beneficial bioactivities including anti-obesity and hypocholesterol effects. This study aimed to assess the effect of the administration of protein hydrolysates from anchovy waste (APH) for 12 weeks on attenuated high-fat diet-induced MAFLD in apolipoprotein E-knockout mice (ApoE–/–). Thirty ApoE–/– mice were divided into two groups (n = 15/group) and fed a high-fat diet (HFD), with and without the addition of 10% (w/w) APH. After 12 weeks, serum and hepatic lipid profiles, hepatic enzyme activities, liver histology and immunohistochemistry were analyzed to assess hepatic steatosis, inflammation and fibrosis. Twelve-weeks on a 10% (w/w) APH diet reduces total cholesterol and triglyceride serum levels, hepatic enzyme activity and hepatic triacylglycerol content (p < 0.0001), and results in a reduction in hepatic fat accumulation and macrophage recruitment (p < 0.0001). The results suggest that a 10% APH diet has an anti-obesity effect, with an improvement in lipid metabolism, hepatic steatosis and liver injury as a result of a high-fat diet. Protein hydrolysates from fish waste may represent an efficient nutritional strategy in several diseases, and their use as nutraceuticals is worthy of future investigation.

Nutritional Characteristics of a Peruvian Anchovy (Engraulis ringens) Protein Concentrate

ABSTRACT The aim of the present study is to present a new anchovy protein concentrate and compare it to other similar fishery products. A 76.4% protein concentrate was prepared from fresh Peruvian anchoveta (Engraulis ringens) (APC) by means of acid treatment with 1% citric acid, solubilization with 1% sodium chloride, and isoelectric precipitation processes. Remaining fat was 5.2%, and 0.51 Aw indicates good stability. When APC was rehydrated for 10 minutes, it had a 3.5 initial weight increase. It provided 17.8% protein of very high nutritional quality, with only histidine content deficient for children 1–2 years old. Fat content in APC offered 33.4% EPA+DHA. No changes in the essential/non-essential amino acids (EAA/non-EAA), polyunsaturated/saturated fatty acids (PUFA/SFA), or w6/w3 ratios were observed as a result of the process in the raw material. Because of the high sodium content, a 25 g portion is recommended to fulfill nutritional FAO recommendations. APC can be considered as an important source of calcium (30% adults and 35% children requirements), phosphorous (30% adults and 52% children requirements), and iron content (approximately 20% adults and children needs). APC is an important high-quality protein, providing w-3 fatty acids, calcium, phosphorous, and iron.

PREPARATION AND CHARACTERIZATION OF ANCHOVY (STOLEPHORUS SP) PROTEIN CONCENTRATE NANOPARTICLE USING IONIC GELATION METHOD

Objective: Nanoparticle preparation can increase the bioavailability of anchovy protein. This study aims to optimize and characterize anchovy protein concentrate nanoparticle based on volume comparison of chitosan-TPP (Tripolyphosphate) matrix in forming the size of anchovy protein concentrate nanoparticle with ionic gelation method.&#x0D; Methods: Fresh anchovy was extracted by an acid-base method using solvents of NaOH and HCl to get protein concentrate. After that, it was conducted by formulating anchovy protein concentrate into a mixture of chitosan-TPP matrix with volume ratio which was divided into 5 series of variations, namely F1 without matrix, F2 1:1, F3 1:3, F4 1:5 and F5 1:7 ratio of chitosan to TPP, then formed matrix was determined the value of the encapsulation efficiency and characterized using a particle size analysis tool.&#x0D; Results: The results showed that the optimum volume comparison of the chitosan-TPP matrix in forming nanoparticle of anchovy protein concentrate was ratio 1:7 because this has a matrix encapsulation efficiency value of 99.11%, a small particle concentrate size of 687.26 nm, a good polydispersity index value of 0.406 and a value of zeta potential of+12.9 mV.&#x0D; Conclusion: The optimum formula of the chitosan-TPP matrix volume ratio in forming anchovy protein concentrate nanoparticle is F5 with the ratio of chitosan-TPP matrix mixture 1:7.

Effects of Maillard reaction on bioactivities promotion of anchovy protein hydrolysate: The key role of MRPs and newly formed peptides with basic and aromatic amino acids

Among multiple ways to ameliorate Alzheimer's disease, antioxidant and acetylcholinesterase (AChE) inhibition are considered as two primary approaches. Previous studies have indicated that anchovy protein hydrolysate (APH) exhibited strong antioxidant and AChE inhibitory capacities, and Maillard reaction could significantly (p < 0.05) improve its bioactivities. The objective of this study was to determine the relationship between chemical compounds and bioactivities (antioxidant and AChE inhibitory capacities) by analyzing the main components of APH with or without Maillard reaction. GC-MS and UPLC-MS/MS results showed that newly formed heterocyclic components and rearrangement peptides with basic or aromatic amino acids in low molecular weight fraction during Maillard reaction might have contribution to the greater ABTS·+ scavenging and AChE inhibitory capacities of Maillard reaction products (MRPs) derived from APH and ribose (APH-M). Therefore, Maillard reaction could raise the bioactivity of hydrolysates which has important guiding significance for the production of polypeptide health care products.

Effect of Organic Acids on Physical-Mechanical and Antifungicidal Properties of Anchovy Protein Films

ABSTRACTThe use of active biodegradable films from renewable sources like anchovy, with the incorporation of organic acids, such as sorbic acid (SA) and benzoic acid (BA), is an alternative to decrease environmental pollution and minimize the development of fungi in foods. This study investigated the respective influences of SA and BA at 0, 0.50, 0.75, and 1.50% on the growth of Aspergillus flavus and Rhizopus oryzae and on Argentine anchovy protein film properties (L*, a*, b*, tensile strength, elongation at break, solubility in water, water vapor permeability, and Fourier transform infrared spectroscopy (FTIR)). Results showed that the incorporation of SA or BA at different levels significantly increased the water vapor permeability, water solubility, and elongation at break of the films, but decreased the tensile strength and L* parameter (p < 0.05). FTIR evidenced an interaction between the BA and Argentine protein isolate. Also, the influences of the SA or BA concentration on antifungal activity was observed. The greatest effectiveness was obtained for films containing SA against R. oryzae compared to BA film treatments. In conclusion, the Argentine anchovy protein films could retain their antifungal property by incorporation of these organic acids.

Neuroprotective Effects of Acetylcholinesterase Inhibitory Peptides from Anchovy (Coilia mystus) against Glutamate-Induced Toxicity in PC12 Cells.

Ameliorations of cholinergic system dysfunction and oxidative stress in neurodegenerative diseases were main approaches to improve memory disorder. Our previous investigation showed that anchovy protein hydrolysate (APH) could attenuate scopolamine-induced memory deficits in mice by regulating acetylcholinesterase (AChE) activity. Therefore, peptides with AChE inhibitory activity in APH were explored and identified in this study, and their possible neuroprotective mechanisms on glutamate induced apoptosis in PC12 were also elucidated. Two peptides with strong AChE inhibitory capacity were identified as Pro-Ala-Tyr-Cys-Ser (PAYCS) and Cys-Val-Gly-Ser-Tyr (CVGSY) by ultraperformance liquid chromatography coupled with tandem mass spectrometry. The AChE inhibitory was 23.68 ± 0.97% and 6.08 ± 0.41%, respectively. Treatment with PAYCS and CVGSY could significantly (p < 0.05) increase cells viability, reduce lactate dehydrogenase release, reactive oxygen species (ROS) production, malondialdehyde content, and the ratio of Bax/Bcl-2 of glutamate-induced apoptosis PC12 cells (82.78 ± 6.58 and 109.94 ± 7.16% of control, respectively) as well as increase superoxide dismutase and GSH-px activities. In addition, both the peptides could inhibit Ca2+ influx but have no effects on mitochondrial membrane potential. Results indicated that AChE inhibitory peptides (PAYCS and CVGSY) possibly protected the PC12 cells against glutamate-induced apoptosis via inhibiting ROS production and Ca2+ influx. PAYCS and CVGSY might be considered as nutraceuticals for alleviating memory deficits.

Antioxidant and anti‐acetylcholinesterase activities of anchovy (Coilia mystus) protein hydrolysates and their memory‐improving effects on scopolamine‐induced amnesia mice

SummaryAnchovy protein hydrolysates (APHs) were prepared through hydrolysis for 2, 4 or 8 h (APH‐2, APH‐4 and APH‐8, respectively). The chemical analyses, in vitro assessments [antioxidant activity and acetylcholinesterase (AchE) inhibitory activity] and in vivo mice tests were evaluated. Results revealed that APH‐8 exhibited the strongest reducing power and AchE inhibitory capacity (IC50 = 159.76 ± 0.03 mg mL−1), which may be due to its specific amino acid composition and newly formed peptides. In addition, AchE inhibitory kinetics of amino acids suggested that lysine was featured of both competitive and noncompetitive inhibitors. Furthermore, the results of in vivo study showed that all APHs exhibited memory‐improving action on scopolamine‐induced amnesia mice especially, APH‐8, indicating that anchovy protein is a potential source for health‐promoting peptides.

Biochemical and Sensory Changes of Low-Salt Anchovy (Engraulis japonicus) Sauce Prepared by a Novel Technique

ABSTRACTUsing a process spanning less than 10 days, a novel technique combining two-stage autolysis and external koji fermentation for the rapid preparation of low-salt anchovy sauce is presented. High-efficiency autolysis of anchovy was achieved by two-stage autolysis, and the second-stage autolysis exhibited a significantly positive effect on the improvement of hydrolysis yield and recovery of anchovy proteins. During the external koji fermentation of hydrolysates from two-stage autolysis, the optimum amounts of added water and salt were determined to be 20% and 8.0%, respectively. By investigating the effect of fermentation time on the biochemical and sensory changes of anchovy sauce, the optimal fermentation time was 8 days. The prepared anchovy sauce had an umami taste, was rich in free glutamate, and contained taurine and γ-aminobutyric acid. In addition, it contained 0.890 g/100 mL amino acid nitrogen (AAN), 1.886 g/100 mL total nitrogen, and 9.76% NaCl. Meanwhile, the recovery of total nitrogen (RTN) was 63.89%.

A Study on the Salty Enhancing Effect in Salad Dressing using Enzymatically Hydrolyzed Anchovy Protein

A Study on the Salty Enhancing Effect in Salad Dressing using Enzymatically Hydrolyzed Anchovy Protein

Salty Taste Enhancing Effect of Enzymatically Hydrolyzed Anchovy Protein in Cooking Application

Salty Taste Enhancing Effect of Enzymatically Hydrolyzed Anchovy Protein in Cooking Application

Selection of Commercial Proteolytic Enzymes for the Preparation of Anchovy Protein Hydrolysates Under Pressurization

Selection of Commercial Proteolytic Enzymes for the Preparation of Anchovy Protein Hydrolysates Under Pressurization

Sensory Characteristics of Enzymatically Hydrolyzed Anchovy Protein by Descriptive Analysis

Sensory Characteristics of Enzymatically Hydrolyzed Anchovy Protein by Descriptive Analysis

Effect of anchovy (Coilia mystus) protein hydrolysate and its Maillard reaction product on combating memory-impairment in mice

Anchovy (Coilia mystus) protein hydrolysate (APH) and its Maillard reaction product (APH-M) with d-ribose were subjected to in vivo mouse behavioral trials and associated with biochemical analyses, in vitro H2O2-stressed PC12 cell assay and inhibition of acetylcholin esterase (AchE) to explore the mechanism of the behavioral trial findings. Results revealed that both APH and APH-M could alleviate undesired alterations on hippocampus ultrastructure of mice, protect PC12 from H2O2-induced oxidative stress, and inhibit AchE activity in vivo and in vitro, suggesting the well-being potential for combating memory-impairment in mice. Moreover, APH-M exhibited much stronger abilities above than APH. The test of acetylcholinergic system indicated that APH-M would improve memory mainly through regulating the AchE activity, while APH through controlling the mRNA expression of ChAT and AchE activity. Therefore, protein hydrolysates from anchovy could possess therapeutic potential for memory deficits, and Maillard reaction might further promote their memory-improving capacity.