Molecular Mechanistic Insights into Dipeptidyl Peptidase-IV Inhibitory Peptides to Decipher the Structural Basis of Activity.

Dipeptidyl peptidase-IV (DPP-IV) inhibiting peptides have attracted increased attention because of their possible beneficial effects on glycemic homeostasis. However, the structural basis underpinning their activities has not been well understood. This study combined computational and in vitro investigations to explore the structural basis of DPP-IV inhibitory peptides. We first superimposed the Xaa-Pro-type peptide-like structures from several crystal structures of DPP-IV ligand-protein complexes to analyze the recognition interactions of DPP-IV to peptides. Thereafter, a small set of Xaa-Pro-type peptides was designed to explore the effect of key interactions on inhibitory activity. The intramolecular interaction of Xaa-Pro-type peptides at the first and third positions from the N-terminus was pivotal to their inhibitory activities. Residue interactions between DPP-IV and residues of the peptides at the fourth and fifth positions of the N-terminus contributed significantly to the inhibitory effect of Xaa-Pro-type tetrapeptides and pentapeptides. Based on the interaction descriptors, quantitative structure-activity relationship (QSAR) studies with the DPP-IV inhibitory peptides resulted in valid models with high R2 values (0.90 for tripeptides; 0.91 for tetrapeptides and pentapeptides) and Q2 values (0.33 for tripeptides; 0.68 for tetrapeptides and pentapeptides). Taken together, the structural information on DPP-IV and peptides in this study facilitated the development of novel DPP-IV inhibitory peptides.

Self-Assembled Food Peptides: Recent Advances and Perspectives in Food and Health Applications.

Self-assembling peptides are rapidly gaining attention as novel biomaterials for food and biomedical applications. Peptides self-assemble when triggered by physical or chemical factors due to their versatile physicochemical characteristics. Peptide self-assembly, when combined with the health-promoting bioactivity of peptides, can also result in a plethora of biofunctionalities of the biomaterials. This perspective highlights current developments in the use of food-derived self-assembling peptides as biomaterials, bioactive nutraceuticals, and potential dual functioning bioactive biomaterials. Also discussed are the challenges and opportunities in the use of self-assembling bioactive peptides in designing biocompatible, biostable, and bioavailable multipurpose biomaterials.

Screening, separation and identification of metal-chelating peptides for nutritional, cosmetics and pharmaceutical applications.

Metal-chelating peptides, which form metal-peptide coordination complexes with various metal ions, can be used as biofunctional ingredients notably to enhance human health and prevent diseases. This review aims to discuss recent insights into food-derived metal-chelating peptides, the strategies set up for their discovery, their study, and identification. After understanding the overall properties of metal-chelating peptides, their production from food-derived protein sources and their potential applications will be discussed, particularly in nutritional, cosmetics and pharmaceutical fields. In addition, the review provides an overview of the last decades of progress in discovering food-derived metal-chelating peptides, addressing several screening, separation and identification methodologies. Furthermore, it emphasizes the methods used to assess peptide-metal interaction, allowing for better understanding of chemical and thermodynamic parameters associated with the formation of peptide-metal coordination complexes, as well as the specific amino acid residues that play important roles in the metal ion coordination.

Relation of amino acid composition, hydrophobicity, and molecular weight with antidiabetic, antihypertensive, and antioxidant properties of mixtures of corn gluten and soy protein hydrolysates.

New mixed Alcalase-hydrolysates were developed using corn gluten meal (CP) and soy protein (SP) hydrolysates, namely CPH, SPH, SPH30:CPH70, SPH70:CPH30, and SPH50:CPH50. Amino acid profile, surface hydrophobicity (H 0), molecular weight (MW) distribution, antioxidant activity, angiotensin-converting enzyme (ACE), α-amylase, and α-glucosidase inhibitory activities, and functional characteristics of hydrolysates were determined. Hydrolysis changed the amount of hydrophilic and hydrophobic amino acid composition and significantly increased the H 0 values of hydrolysates, especially for CPH. The DPPH radical scavenging activity (RSA) was higher for CPH, SPH30:CPH70, and SPH50:CPH50 than SPH and SPH70:CPH30. Moreover, SPH, SPH70:CPH30, and SPH50:CPH50 showed lower MW than CPH, and this correlated with the higher hydrophilicity, and ABTS and hydroxyl RSA values obtained for SPH and the mixed hydrolysates with predominantly SPH. SPH70:CPH30 exhibited higher ACE, α-glucosidase, and α-amylase inhibitory activities among all samples due to its specific peptides with high capacity to interact with amino acid residues located at the enzyme active site and also low binding energy. At 15% degree of hydrolysis, both SPH and CPH showed enhanced solubility at pH 4.0, 7.0 and 9.0, emulsifying activity, and foaming capacity. Taken together, SPH70:CPH30 displayed strong antioxidant, antihypertensive, and antidiabetic attributes, emulsifying activity and stability indexes, and foaming capacity and foaming stability, making it a promising multifunctional ingredient for the development of functional food products.

In Silico Exploration of Metabolically Active Peptides as Potential Therapeutic Agents against Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is regarded as a fatal neurodegenerative disease that is featured by progressive damage of the upper and lower motor neurons. To date, over 45 genes have been found to be connected with ALS pathology. The aim of this work was to computationally identify unique sets of protein hydrolysate peptides that could serve as therapeutic agents against ALS. Computational methods which include target prediction, protein-protein interaction, and peptide-protein molecular docking were used. The results showed that the network of critical ALS-associated genes consists of ATG16L2, SCFD1, VAC15, VEGFA, KEAP1, KIF5A, FIG4, TUBA4A, SIGMAR1, SETX, ANXA11, HNRNPL, NEK1, C9orf72, VCP, RPSA, ATP5B, and SOD1 together with predicted kinases such as AKT1, CDK4, DNAPK, MAPK14, and ERK2 in addition to transcription factors such as MYC, RELA, ZMIZ1, EGR1, TRIM28, and FOXA2. The identified molecular targets of the peptides that support multi-metabolic components in ALS pathogenesis include cyclooxygenase-2, angiotensin I-converting enzyme, dipeptidyl peptidase IV, X-linked inhibitor of apoptosis protein 3, and endothelin receptor ET-A. Overall, the results showed that AGL, APL, AVK, IIW, PVI, and VAY peptides are promising candidates for further study. Future work would be needed to validate the therapeutic properties of these hydrolysate peptides by in vitro and in vivo approaches.

Transepithelial transport and cellular mechanisms of food-derived antioxidant peptides.

Considering the involvement of oxidative stress in the etiology of many non-communicable diseases, food-derived antioxidant peptides (FDAPs) are strong candidates for nutraceutical development for disease prevention and management. This paper reviews current evidence on the transepithelial transport and cellular mechanisms of antioxidant activities of FDAPs. Several FDAPs have multiple health benefits such as anti-inflammatory and anti-photoaging activities, in addition to antioxidant properties through which they protect cellular components from oxidative damage. Some FDAPs have been shown to permeate the intestinal epithelium, which could facilitate their bioavailability and physiological bioactivities. Molecular mechanisms of FDAPs include suppression of oxidative stress as evidenced by reduction in intracellular reactive oxygen species production, lipid peroxidation and apoptotic protein activation as well as increase in antioxidant defense mechanisms (enzymatic and non-enzymatic). Since many FDAPs have demonstrated promising antioxidant activity, future investigation should focus on further elucidation of molecular mechanisms and human studies to explore their practical application for the prevention and management of oxidative stress-related diseases.

Preparation, pungency and bioactivity of gingerols from ginger (Zingiber officinale Roscoe): a review.

Ginger has been widely used for different purposes, such as condiment, functional food, drugs, and cosmetics. Gingerols, the main pungent component in ginger, possess a variety of bioactivities. To fully understand the significance of gingerols in the food and pharmaceutical industry, this paper first recaps the composition and physiochemical properties of gingerols, and the major extraction and synthesis methods. Furthermore, the pungency and bioactivity of gingerols are reviewed. In addition, the food application of gingerols and future perspectives are discussed. Gingerols, characterized by a 3-methoxy-4-hydroxyphenyl moiety, are divided into gingerols, shogaols, paradols, zingerone, gingerdiones and gingerdiols. At present, gingerols are extracted by conventional, innovative, and integrated extraction methods, and synthesized by chemical, biological and in vitro cell synthesis methods. Gingerols can activate transient receptor potential vanilloid type 1 (TRPV1) and induce signal transduction, thereby exhibiting its pungent properties and bioactivity. By targeted mediation of various cell signaling pathways, gingerols display potential anticancer, antibacterial, blood glucose regulatory, hepato- and renal-protective, gastrointestinal regulatory, nerve regulatory, and cardiovascular protective effects. This review contributes to the application of gingerols as functional ingredients in the food and pharmaceutical industry.

Osteo-modulatory dietary proteins and peptides: A concise review.

The integrity of the bone is dependent on the strict balance between osteoclastogenesis and osteoblastogenesis, and any imbalance results in bone diseases. Dietary proteins (DP) have been shown to promote osteogenesis while inhibiting bone resorption in cultured osteoblasts, and in animal models of bone diseases such as ovariectomy, 1α,25-dihydroxy-vitamin D3 (VD3), and prostaglandin E2 (PGE2)-induced bone resorption. Hydrolysis of some of these DPs with osteo-modulatory properties has been shown to generate hydrolysates with bioactive peptides that exhibit higher osteo-modulatory properties in comparison to intact (parent) proteins. The higher bioactivity of the isolated peptides and protein hydrolysates compared to intact proteins indicates that the osteo-modulatory properties are dependent on the degree of exposure of the functional groups of amino acid residues involved in target interaction. This review provides an overview of the preparation of DP and select peptides with osteo-modulatory properties, and summarizes the potential underlying mechanisms of action through which the bioactive peptides help maintain bone health. PRACTICAL APPLICATIONS: Bone diseases such as osteoporosis (OP), osteoarthritis (OA), bone cancer (BC), and others have negative impacts on the quality of life, especially in older women after menopause. Current drugs used in treating many bone diseases such as bisphosphonates, anabolic steroids, and selective estrogen receptor modulators have been limited by worrisome adverse effects such as organ toxicity, increased risk of cancer, and cardiovascular abnormalities, and gastrointestinal discomfort. There is growing scientific evidence that certain multifunctional dietary proteins and bioactive peptides may positively modulate bone health by modifying risk factors for bone diseases including inflammation, oxidative stress, hyperlipidemia, and hyperglycemia.

TNKPVI, a Putative Bioaccessible Pharmacophore of Anti-Inflammatory Potato Patatin-Derived Decapeptide DIKTNKPVIF.

Potato protein-derived decapeptide DIKTNKPVIF exerted anti-inflammatory activity in animal models when delivered via intragastric gavage and intraperitoneal injection. However, DIKTNKPVIF is susceptible to hydrolysis in the digestive tract, which will decrease its bioaccessibility and possibly bioactivity. In this study, the anti-inflammatory activity of fragments generated from in silico gastrointestinal enzymatic hydrolysis of DIKTNKPVIF was investigated using the human monocytic (THP-1) cell line. The simulated digestion by pepsin and trypsin released four fragments, DIKTNKPVI, TNKPVIF, DIK and TNKPVI. The peptides lacked the cleavage sites of chymotrypsin. All five peptides were predicted to be non-toxic, which was validated using cytotoxicity assay at 0.25-1 mM peptide concentration. However, the peptides were predicted to possess poor pharmacokinetic profiles, including low passive gastrointestinal absorption and blood-brain barrier permeability. TNKPVIF, DIK and TNKPVI significantly reduced the amount of pro-inflammatory interleukin (IL)-6, IL-8 and tumor necrosis factor in lipopolysaccharide-activated THP-1 cells. Notably, the anti-inflammatory activity of fragment TNKPVI was comparable to that of the parent decapeptide while peptide fragment DIKTNKPVI had no apparent effect on the pro-inflammatory cytokines. This highlights the important role of the C-terminal phenylalanine residue of the parent peptide in the bioactivity. Furthermore, given its activity and the absence of cleavage sites of major digestive proteases, TNKPVI could be the biostable and bioaccessible pharmacophore of potato patatin-derived anti-inflammatory decapeptide DIKTNKPVIF.

Anti-COVID-19 potential of Azadirachta indica (Neem) leaf extract.

COVID-19 is caused by infection with the "severe acute respiratory syndrome coronavirus-2″ (i.e., SARS-CoV-2). This is an enveloped virus having a positive sense, single-stranded RNA genome; like the two earlier viruses SARS-CoV and the Middle East respiratory syndrome (MERS) virus. COVID-19 is unique in that, in the severe case, it has the propensity to affect multiple organs, leading to multiple organ distress syndrome (MODS), and causing high morbidity and mortality in the extreme case. In addition, comorbidities like age, cardiovascular disease, diabetes and its complications, obesity, are risk factors for severe COVID-19. It turns out that a most plausible, simple, single explanation for this propensity for MODS is the pivotal involvement of the vascular endothelium (VE). This is a consequence of the fact that the VE seamlessly connects all the entire vascular bed in the body, thus linking all the target organs (heart, lungs, kidney, liver, brain) and systems. Infection with SARS-CoV-2 leads to hyper-inflammation yielding uncontrolled production of a mixture of cytokines, chemokines, reactive oxygen species, nitric oxide, oxidative stress, acute phase proteins (e.g., C-reactive protein), and other pro-inflammatory substances. In the extreme case, a cytokine storm is created. Displacement of the virus bound to the VE, and/or inhibition of binding of the virus, would constitute an effective strategy for preventing COVID-19. In this regard, the acetone-water extract of the leaf of the Neem (Azadirachta indica) plant has been known to prevent the adherence of malaria parasitized red blood cells (pRBCs) to VE; prevent cytoadherence of cancer cells in metastasis; and prevent HIV from invading target T lymphocytes. We therefore hypothesize that this Neem leaf acetone-water extract will prevent the binding of SARS-CoV-2 to the VE, and therefore be an effective therapeutic formulation against COVID-19. It is therefore advocated herein that this extract be investigated through rigorous clinical trials for this purpose. It has the advantages of being (i) readily available, and renewable in favor of the populations positioned to benefit from it; (ii) simple to prepare; and (iii) devoid of any detectable toxicity.

A review on the techno-functional, biological, and health-promoting properties of hempseed-derived proteins and peptides.

Protein-energy malnutrition is a global challenge that demands urgent attention, especially with the increasing population growth and unmatched food security plans. One strategy is to expand the list of protein sources, such as neglected and underutilized crops, with high protein content. A good number of plant proteins, in addition to their nutritional benefits, exert therapeutic properties as seen in seeds derived from legumes and emerging sources such as hemp. In this review, the transepithelial transport, functional, and biological properties of hempseed proteins (HSPs) and peptides were discussed. The review also described the potential safety issues of incorporating hempseeds in food products. Due to the multitargeted effects of hempseed-derived proteins and their peptides against many chronic diseases, and their functional properties, current knowledge shows that hempseed has tremendous potential for functional food and nutraceutical applications. PRACTICAL APPLICATIONS: The alarming rate of malnutrition and the attendant health consequences demand that underexploited nutrient-rich crops should be incorporated as part of our common dietary sources. Among these crops, hempseed is gaining attention as an emerging source of proteins and peptides with promising potential in prevention and management of chronic diseases such as diabetes, hypertension, cancer, hypercholesterolemia, obesity, and diseases whose etiology involves oxidative stress and inflammation. Fortunately, a growing body of research evidence is demonstrating that hempseed is a reservoir of proteins and peptides with nutraceutical potentials for curbing life-threatening diseases.

Maillard-Type Protein-Polysaccharide Conjugates and Electrostatic Protein-Polysaccharide Complexes as Delivery Vehicles for Food Bioactive Ingredients: Formation, Types, and Applications.

Due to their combination of featured properties, protein and polysaccharide-based carriers show promising potential in food bioactive ingredient encapsulation, protection, and delivery. The formation of protein-polysaccharide complexes and conjugates involves non-covalent interactions and covalent interaction, respectively. The common types of protein-polysaccharide complex/conjugate-based bioactive ingredient delivery systems include emulsion (conventional emulsion, nanoemulsion, multiple emulsion, multilayered emulsion, and Pickering emulsion), microcapsule, hydrogel, and nanoparticle-based delivery systems. This review highlights the applications of protein-polysaccharide-based delivery vehicles in common bioactive ingredients including polyphenols, food proteins, bioactive peptides, carotenoids, vitamins, and minerals. The loaded food bioactive ingredients exhibited enhanced physicochemical stability, bioaccessibility, and sustained release in simulated gastrointestinal digestion. However, limited research has been conducted in determining the in vivo oral bioavailability of encapsulated bioactive compounds. An in vitro simulated gastrointestinal digestion model incorporating gut microbiota and a mucus layer is suggested for future studies.

Disaggregation of Islet Amyloid Polypeptide Fibrils as a Potential Anti-Fibrillation Mechanism of Tetrapeptide TNGQ.

Islet amyloid polypeptide (IAPP) fibrillation has been commonly associated with the exacerbation of type 2 diabetes prognosis. Consequently, inhibition of IAPP fibrillation to minimize ß-cell cytotoxicity is an important approach towards ß-cell preservation and type 2 diabetes management. In this study, we identified three tetrapeptides, TNGQ, MANT, and YMSV, that inhibited IAPP fibrillation. Using thioflavin T (ThT) fluorescence assay, circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), and molecular docking, we evaluated the potential anti-fibrillation mechanism of the tetrapeptides. ThT fluorescence kinetics and microscopy as well as transmission electron microscopy showed that TNGQ was the most effective inhibitor based on the absence of normal IAPP fibrillar morphology. CD spectroscopy showed that TNGQ maintained the α-helical conformation of monomeric IAPP, while DLS confirmed the presence of varying fibrillation species. Molecular docking showed that TNGQ and MANT interact with monomeric IAPP mainly by hydrogen bonding and electrostatic interaction, with TNGQ binding at IAPP surface compared to YMSV, which had the highest docking score, but interact mainly through hydrophobic interaction in IAPP core. The highly polar TNGQ was the most active and appeared to inhibit IAPP fibrillation by disaggregation of preformed IAPP fibrils. These findings indicate the potential of TNGQ in the development of peptide-based anti-fibrillation and antidiabetic nutraceuticals.

Inhibition of low-density lipoprotein oxidation, antioxidative and bile acid-binding capacities of hydrolyzed proteins from carbohydrase-treated oat bran.

This study investigated the valorization of oat bran and the use of its proteins to generate polypeptides with antioxidant and bile acid-binding properties. Ten protein hydrolysates were prepared by treating cellulase (CPI) or Viscozyme (VPI) protein isolates with five proteases. VPI-pepsin was the best peroxyl radical scavenger (497 ± 6-µM Trolox equivalents [TE]/g) while VPI-Flavourzyme quenched hydroxyl radicals (28 ± 0.6) and VPI-pepsin superoxide anion radicals (45.3 ± 6.6%). Hydrolysates, except those produced with pepsin, dose-dependently chelated iron whereas VPI-Protamex had the best copper-chelating capacity (59.83 ± 1.40%). These antioxidative capacities were important in preventing by 50% in vitro copper-induced oxidation of human low-density lipoprotein. Furthermore, due to their aromatic amino acid contents and hydrophobicity, the hydrolysates bound up to 46.3% the bile acids taurodeoxycholate and taurocholate. PRACTICAL APPLICATIONS: The presence of oxidants in foods can damage food molecules and decrease their quality. They are also known to increase the risk of developing chronic conditions like cardiovascular disease. Finding new antioxidant molecules are therefore useful in the management of chronic diseases. Data from this work showed that hydrolyzed oat bran proteins can be useful in stabilizing commercial oil as they reduced the oxidation of peanut oil. Additionally, the protein hydrolysates not only prevented the oxidation of linoleic, a common component of both vegetable oils and biological cell membranes, they also inhibited the oxidation of human LDL cholesterol and chelated bile acids. These hydrolysates can then be further explored as multifunctional ingredients for the development of stable functional food products with potential beneficial effects on the cardiovascular system.

Buckwheat proteins: functionality, safety, bioactivity, and prospects as alternative plant-based proteins in the food industry.

The need for protein in human nutrition is rapidly increasing because of the increasing world population and consumer preference for high-protein foods. Plant proteins are gaining attention as sustainable means of meeting the global protein need due to their lower carbon footprint. Nonetheless, the food industry has neglected or underutilized many plant proteins, including buckwheat protein. Buckwheat is a pseudocereal and its groats contain beneficial components such as proteins, dietary fiber, vitamins, and bioactive polyphenols. The protein quality of buckwheat seeds varies between the tartary and common buckwheat types; both are gluten-free and contain considerable amount of indispensable amino acids. This review provides a detailed discussion on the profile, amino acid composition, digestibility, allergenicity, functional properties, and bioactivity of buckwheat proteins. Prospects of processing buckwheat for improving protein digestibility and deactivating allergenic epitopes were also discussed. Based on the literature, buckwheat protein has a tremendous potential for utilization in structuring food products and developing peptide-based functional foods for disease prevention. Future research should develop new processing technologies for further improvement of the quality and functional properties of buckwheat protein in order to facilitate its utilization as an alternative plant-based protein toward meeting the global protein supply.

Inhibition of Islet Amyloid Polypeptide Fibrillation by Structurally Diverse Phenolic Compounds and Fibril Disaggregation Potential of Rutin and Quercetin.

The influence of 12 food-derived phenolic compounds on islet amyloid polypeptide (IAPP) fibrillation was investigated. Results from thioflavin T assay demonstrated that gallic acid, caffeic acid, and rutin and its aglycone, quercetin, inhibited IAPP fibrillation at 1:0.5, 1:1, and 1:2 IAPP-phenolic molar ratios. Circular dichroism and dynamic light scattering at the 1:1 IAPP-phenolic ratio confirmed the inhibition of fibril formation. Rutin and quercetin increased the lag time by 90 and 6%, and the relative α-helix content by 63 and 48%, respectively. Gallic acid decreased the elongation rate by 30%, whereas caffeic acid decreased the maximum fluorescence intensity by 65%. Furthermore, fluorescence microscopy and transmission electron microscopy (TEM) showed IAPP fibril morphologies indicative of fibrillation reduction by the compounds. Molecular docking and TEM showed that rutin and quercetin disaggregated preformed IAPP fibrils potentially through fibrillar-monomeric equilibrium shifts. These findings demonstrate important structural features of phenolic compounds for disaggregating IAPP fibrils or inhibiting their formation.

Production, Purification, and Potential Health Applications of Edible Seeds' Bioactive Peptides: A Concise Review.

Edible seeds play a significant role in contributing essential nutritional needs and impart several health benefits to improve the quality of human life. Previous literature evidence has confirmed that edible seed proteins, their enzymatic hydrolysates, and bioactive peptides (BAPs) have proven and potential attributes to ameliorate numerous chronic disorders through the modulation of activities of several molecular markers. Edible seed-derived proteins and peptides have gained much interest from researchers worldwide as ingredients to formulate therapeutic functional foods and nutraceuticals. In this review, four main methods are discussed (enzymatic hydrolysis, gastrointestinal digestion, fermentation, and genetic engineering) that are used for the production of BAPs, including their purification and characterization. This article's main aim is to provide current knowledge regarding several health-promoting properties of edible seed BAPs in terms of antihypertensive, anti-cancer, antioxidative, anti-inflammatory, and hypoglycemic activities.

Zanthoxylum Species: A Review of Traditional Uses, Phytochemistry and Pharmacology in Relation to Cancer, Infectious Diseases and Sickle Cell Anemia.

The health benefits and toxicity of plant products are largely dependent on their secondary metabolite contents. These compounds are biosynthesized by plants as protection mechanisms against environmental factors and infectious agents. This review discusses the traditional uses, phytochemical constituents and health benefits of plant species in genus Zanthoxylum with a focus on cancer, microbial and parasitic infections, and sickle cell disease as reported in articles published from 1970 to 2021 in peer-reviewed journals and indexed in major scientific databases. Generally, Z. species are widely distributed in Asia, America and Africa, where they are used as food and for disease treatment. Several compounds belonging to alkaloids, flavonoids, terpenoids, and lignans, among others have been isolated from Z. species. This review discusses the biological activities reported for the plant species and their phytochemicals, including anticancer, antibacterial, antifungal, antiviral, anti-trypanosomal, antimalarial and anti-sickling properties. The safety profiles and suggestions for conservation of the Z. species were also discussed. Taken together, this review demonstrates that Z. species are rich in a wide range of bioactive phytochemicals with multiple health benefits, but more research is needed towards their practical application in the development of functional foods, nutraceuticals and lead compounds for new drugs.

Anti-Salmonella Activity and Peptidomic Profiling of Peptide Fractions Produced from Sturgeon Fish Skin Collagen (Huso huso) Using Commercial Enzymes.

This study investigated peptide fractions from fish skin collagen for antibacterial activity against Escherichia coli and Salmonella strains. The collagen was hydrolyzed with six commercial proteases, including trypsin, Alcalase, Neutrase, Flavourzyme, pepsin and papain. Hydrolyzed samples obtained with trypsin and Alcalase had the largest number of small peptides (molecular weight <10 kDa), while the hydrolysate produced with papain showed the lowest degree of hydrolysis and highest number of large peptides. Four hydrolysates were found to inhibit the growth of the Gram-negative bacteria, with papain hydrolysate showing the best activity against E. coli, and Neutrase and papain hydrolysates showing the best activity against S. abony; hydrolysates produced with trypsin and pepsin did not show detectable antibacterial activity. After acetone fractionation of the latter hydrolysates, the peptide fractions demonstrated enhanced dose-dependent inhibition of the growth (colony-forming units) of four Salmonella strains, including S. abony (NCTC 6017), S. typhimurium (ATCC 13311), S. typhimurium (ATCC 14028) and S. chol (ATCC 10708). Shotgun peptidomics analysis of the acetone fractions of Neutrase and papain hydrolysates resulted in the identification of 71 and 103 peptides, respectively, with chain lengths of 6-22 and 6-24, respectively. This work provided an array of peptide sequences from fish skin collagen for pharmacophore identification, structure-activity relationship studies, and further investigation as food-based antibacterial agents against pathogenic microorganisms.

A Concise Review of Current In Vitro Chemical and Cell-Based Antioxidant Assay Methods.

Antioxidants remain interesting molecules of choice for suppression of the toxic effects of free radicals in foods and human systems. The current practice involves the use of mainly synthetic molecules as potent antioxidant agents. However, due to the potential negative impact on human health, there is an intensive effort within the research community to develop natural alternatives with similar antioxidant efficacy but without the negative side effects of synthetic molecules. Still, the successful development of new molecules depends on the use of reliable chemical or cell culture assays to screen antioxidant properties. Chemical antioxidant assays include the determination of scavenging ability against free radicals such as DPPH, superoxide anion radicals, hydroxyl radicals, hydrogen peroxide, and nitric oxide. Other antioxidant tests include the ability of compounds to bind and sequester prooxidant metal cations, reduce ferric iron, and attenuate the rate of lipid oxidation. Ex vivo tests utilize cell cultures to confirm entry of the molecules into cells and the ability to quench synthetic intracellular free radicals or to stimulate the increased biosynthesis of endogenous antioxidants. In order to assist researchers in their choice of antioxidant evaluation methods, this review presents background scientific information on some of the most commonly used antioxidant assays with a comparative discussion of the relevance of published literature data to food science and human nutrition applications.