Insights into Molecular Interactions between a GAPDH-Related Fish Antimicrobial Peptide, Analogs Thereof, and Bacterial Membranes.

Interactions between SJGAP (skipjack tuna GAPDH-related antimicrobial peptide) and four analogs thereof with model bacterial membranes were studied using Fourier-transform infrared spectroscopy (FTIR) and molecular dynamics (MD) simulations. MD trajectory analyses showed that the N-terminal segment of the peptide analogs has many contacts with the polar heads of membrane phospholipids, while the central α helix interacts strongly with the hydrophobic core of the membranes. The peptides also had a marked influence on the wave numbers associated with the phase transition of phospholipids organized as liposomes in both the interface and aliphatic chain regions of the infrared spectra, supporting the interactions observed in the MD trajectories. In addition, interesting links were found between peptide interactions with the aliphatic chains of membrane phospholipids, as determined by FTIR and from the MD trajectories, and the membrane permeabilization capacity of these peptide analogs, as previously demonstrated. To summarize, the combined experimental and computational efforts have provided insights into crucial aspects of the interactions between the investigated peptides and bacterial membranes. This work thus makes an original contribution to our understanding of the molecular interactions underlying the antimicrobial activity of these GAPDH-related antimicrobial peptides from Scombridae.

Physicochemical properties of wild and cultivated Saccharina latissima macroalgae harvested in the Canadian boreal-subarctic transition zone.

Saccharina latissima is a brown seaweed used as a food ingredient. The aim of this work was to study possible differences between S. latissima chemical composition, color, mode of cultivation, harvesting period and site and its environmental conditions. Water temperature, salinity, radiation, and fluorescence were monitored in each harvesting site. Chemical composition of S. latissima varied greatly with period and site, with a high content of carbohydrates and ash. Crude protein content varied from 3.7 % to 12.8 %, with a higher concentration observed in wild samples harvested in Bas-St. Laurent (11.1-12.8 %). Cultivated seaweed also presented a high crude protein (12.2 %) and ash (52 % against 27 % in wild samples) concentrations, but crude fiber and carbohydrates concentrations were lower, reaching up to 2.7 and 1.9-fold, respectively, than those in wild seaweeds. S. latissima presented a more intense yellow color in June. A trend of darker and more green-colored seaweeds when cultivated in the end of summer was confirmed. Our results suggest that variations in chemical components and chromaticity of this species are probably affected by complex interactions of environmental conditions.

Assessing the Activity under Different Physico-Chemical Conditions, Digestibility, and Innocuity of a GAPDH-Related Fish Antimicrobial Peptide and Analogs Thereof.

The antimicrobial activity of SJGAP (skipjack tuna GAPDH-related antimicrobial peptide) and four chemical analogs thereof was determined under different physicochemical conditions, including different pH values, the presence of monovalent and divalent cations, and after a heating treatment. The toxicity of these five peptides was also studied with hemolytic activity assays, while their stability under human gastrointestinal conditions was evaluated using a dynamic in vitro digestion model and chromatographic and mass spectrometric analyses. The antibacterial activity of all analogs was found to be inhibited by the presence of divalent cations, while monovalent cations had a much less pronounced impact, even promoting the activity of the native SJGAP. The peptides were also more active at acidic pH values, but they did not all show the same stability following a heat treatment. SJGAP and its analogs did not show significant hemolytic activity (except for one of the analogs at a concentration equivalent to 64 times that of its minimum inhibitory concentration), and the two analogs whose digestibility was studied degraded very rapidly once they entered the stomach compartment of the digestion model. This study highlights for the first time the characteristics of antimicrobial peptides from Scombridae or homologous to GAPDH that are directly related to their potential clinical or food applications.

Impact of Harvest Month and Drying Process on the Nutritional and Bioactive Properties of Wild Palmaria palmata from Atlantic Canada.

The macroalga Palmaria palmata could be a sustainable and nutritional food resource. However, its composition may vary according to its environment and to processing methods used. To investigate these variations, wild P. palmata from Quebec were harvested in October 2019 and June 2020, and dried (40 °C, ≃5 h) or stored as frozen controls (-80 °C). The chemical (lipids, proteins, ash, carbohydrates, fibers), mineral (I, K, Na, Ca, Mg, Fe), potential bioactive compound (carotenoids, polyphenols, ß-carotene, α-tocopherol) compositions, and the in vitro antioxidant activity and angiotensin-converting enzyme (ACE) inhibition potential of water-soluble extracts were determined. The results suggested a more favorable macroalgae composition in June with a higher content of most nutrients, minerals, and bioactive compounds. October specimens were richer only in carbohydrates and carotenoids. No significant differences in antioxidant or anti-ACE inhibitory activities were found between the two harvest months. The drying process did not significantly impact the chemical and mineral compositions, resulting in only small variations. However, drying had negative impacts on polyphenols and anti-ACE activities in June, and on carotenoids in October. In addition, a concentration effect was observed for carotenoids, ß-carotene and α-tocopherol in June. To provide macroalgae of the highest nutritional quality, the drying process for June specimens should be selected.

Variation of the Nutritional Composition and Bioactive Potential in Edible Macroalga Saccharina latissima Cultivated from Atlantic Canada Subjected to Different Growth and Processing Conditions.

Macroalgae are a new food source in the Western world. The purpose of this study was to evaluate the impact of harvest months and food processing on cultivated Saccharina latissima (S. latissima) from Quebec. Seaweeds were harvested in May and June 2019 and processed by blanching, steaming, and drying with a frozen control condition. The chemical (lipids, proteins, ash, carbohydrates, fibers) and mineral (I, K, Na, Ca, Mg, Fe) compositions, the potential bioactive compounds (alginates, fucoidans, laminarans, carotenoids, polyphenols) and in vitro antioxidant potential were investigated. The results showed that May specimens were significantly the richest in proteins, ash, I, Fe, and carotenoids, while June macroalgae contained more carbohydrates. The antioxidant potential of water-soluble extracts (Oxygen Radical Absorbance Capacity [ORAC] analysis-625 µg/mL) showed the highest potential in June samples. Interactions between harvested months and processing were demonstrated. The drying process applied in May specimens appeared to preserve more S. latissima quality, whereas blanching and steaming resulted in a leaching of minerals. Losses of carotenoids and polyphenols were observed with heating treatments. Water-soluble extracts of dried May samples showed the highest antioxidant potential (ORAC analysis) compared to other methods. Thus, the drying process used to treat S. latissima harvested in May seems to be the best that should be selected.

In Vitro Bioaccessibility of Proteins and Bioactive Compounds of Wild and Cultivated Seaweeds from the Gulf of Saint Lawrence.

Despite the increased interest in macroalgae protein and fibers, little information is available on their bioaccessibility. The application of an in vitro gastrointestinal digestion model to study the degree of disintegration and release of proteins with expressed bioactivities from wild and cultivated Palmaria palmata and Saccharina latissima was proposed in this study. Macroalgae from the Gulf of St Lawrence, Canada, were submitted to digestive transit times of 2 (oral), 60 (gastric) and 120 (duodenal) minutes. Among wild samples, P. palmata had a higher percentage of disintegration, protein release and degree of hydrolysis than S. latissima. While the least digested sample, wild S. latissima, was the sample with the highest antioxidant activity (210 µmol TE g-1), the most digested sample, cultivated P. palmata, presented the highest ability to inhibit the angiotensin-converting enzyme (ACE), reaching 32.6 ± 1.2% at 3 mg mL-1. ACE inhibitory activity increased from 1 to 3 mg mL-1, but not at 5 mg mL-1. Wild samples from both species showed an ACE inhibition around 27.5%. Data suggested that the disintegration of the samples was influenced by their soluble and insoluble fiber contents. Further information on the bioaccessibility and bioactivity of these macroalgae should consider the characterization of digestion products other than protein, as well as the effects of previous product processing.

Beneficial effects of fish and fish peptides on main metabolic syndrome associated risk factors: Diabetes, obesity and lipemia.

The definition of metabolic syndrome (MetS) fairly varies from one to another guideline and health organization. Per description of world health organization, occurrence of hyperinsulinemia or hyperglycemia in addition to two or more factors of dyslipidemia, hypoalphalipoproteinemia, hypertension and or large waist circumference factors would be defined as MetS. Conventional therapies and drugs, commonly with adverse effects, are used to treat these conditions and diseases. Nonetheless, in the recent decades scientific community has focused on the discovery of natural compounds to diminish the side effects of these medications. Among many available bioactives, biologically active peptides have notable beneficial effects on the management of diabetes, obesity, hypercholesterolemia, and hypertension. Marine inclusive of fish peptides have exerted significant bioactivities in different experimental in-vitro, in-vivo and clinical settings. This review exclusively focuses on studies from the recent decade investigating hypoglycemic, hypolipidemic, hypercholesterolemic and anti-obesogenic fish and fish peptides. Related extraction, isolation, and purification methodologies of anti-MetS fish biopeptides are reviewed herein for comparison purposes only. Moreover, performance of biopeptides in simulated gastrointestinal environment and structure-activity relationship along with absorption, distribution, metabolism, and excretion properties of selected oligopeptides have been discussed, in brief, to broaden the knowledge of readers on the design and discovery trends of anti-MetS compounds.Supplemental data for this article is available online at https://doi.org/10.1080/10408398.2022.2052261 .

Identification of Gastrointestinal Digestion Stable Antihypertensive Fish Peptides from Atlantic Mackerel (Scomber scombrus).

High blood pressure has been recognized as one of the main risk factors of cardiometabolic syndrome by health organizations. Hypertension is medicated by various classes of synthetic drugs; however, adverse effects have repeatedly been reported. Moreover, natural alternatives such as fish peptides have been effective in the treatment and prevention of hypertension. The aim of our study was to fractionate and identify antihypertensive peptides. Fractions were produced using different techniques of solid-phase extraction (SPE), pressure-driven ultrafiltration (UF), and electro04dialysis with UF membrane. According to our results, the hydrophobic fraction of SPE (IC50 5 µg) was the most potent anti-angiotensin converting enzyme (ACE) product. Findings of the study suggest that the separation technique plays an important role in the isolation efficiency of antihypertensive biopeptides. Importantly, the hydrophobic fraction's activity was retained through a static model of an in vitro gastrointestinal digestion system. In conclusion, polarity regardless of charge and size was the most important factor for anti-ACE activity of an Atlantic mackerel biopeptide. In addition, the presence of leucine at either of the extremities (C- or N-terminal) and/or leucine-rich motifs could well explain the hypotensive effect of our active fraction.

Determination of the Relationships between the Chemical Structure and Antimicrobial Activity of a GAPDH-Related Fish Antimicrobial Peptide and Analogs Thereof.

The structure-activity relationships and mode of action of synthesized glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-related antimicrobial peptides were investigated. Including the native skipjack tuna GAPDH-related peptide (SJGAP) of 32 amino acid residues (model for the study), 8 different peptide analogs were designed and synthesized to study the impact of net charge, hydrophobicity, amphipathicity, and secondary structure on both antibacterial and antifungal activities. A net positive charge increase, by the substitution of anionic residues or C-terminal amidation, improved the antimicrobial activity of the SJGAP analogs (minimal inhibitory concentrations of 16-64 µg/mL), whereas the alpha helix content, as determined by circular dichroism, did not have a very definite impact. The hydrophobicity of the peptides was also found to be important, especially for the improvement of antifungal activity. Membrane permeabilization assays showed that the active peptides induced significant cytoplasmic membrane permeabilization in the bacteria and yeast tested, but that this permeabilization did not cause leakage of 260 nm-absorbing intracellular material. This points to a mixed mode of action involving both membrane pore formation and targeting of intracellular components. This study is the first to highlight the links between the physicochemical properties, secondary structure, antimicrobial activity, and mechanism of action of antimicrobial peptides from scombrids or homologous to GAPDH.

Algae as a Source of Natural Flavors in Innovative Foods.

As a result of their nutritive values, algae have been used as a food resource for centuries, and there is a growing interest to use them as enrichment ingredients in food products. However, food product acceptance by consumers is strongly linked to their organoleptic properties, especially the aroma, taste, and a combination of the two, flavor. With regard to edible algae, "fresh seashore", "seafood-like", "cucumber green", and "earthy" are descriptors commonly used to define their aromas. Several families of molecules participate in the diversity and peculiarities of algal aromas: pungent sulfur compounds and marine halogenated components but also herbaceous fatty acid derivatives and fruity-floral terpenoids. In both macroalgae (seaweeds) and microalgae, these compounds are studied from a chemistry point of view (identification and quantification) and a sensorial point of view, involving sensorial evaluation by panelists. As a whole food, a food ingredient, or a feed, algae are valued for their nutritional composition and their health benefits. However, because the acceptance of food by consumers is so strongly linked to its sensorial features, studies have been performed to explore the aromas of algae, their impact on food, their evolution through processing, and their ability to produce selected aromas using biotechnology. This review aims at highlighting algal aromas from seaweed and microalgae as well as their use, their handling, and their processing in the food industry.

Valorization of snow crab (Chionoecetes opilio) cooking effluents for food applications.

BACKGROUND: Seafood processing generates significant amounts of solid and liquid waste in the environment. Such waste represents a potential source of high-value biomolecules for food, pharmaceutic and cosmetic applications. There are very few studies on the valorization of wastewaters compared to solid by-products. However, cooking waters are characterized by a high organic polluting load, which could contain valuable molecules such as proteins, pigments and flavor compounds. Snow crab (Chionoecetes opilio) processing is included among the most important processes in Canadian fisheries, although its cooking effluent composition is not well characterized. RESULTS: The present study concentrated and valorized the biomass in snow crab cooking wastewaters for the development of products for food applications. A membrane process was designed and optimized to concentrate the effluents. The chemical composition of the concentrates was analyzed, including characterizing the flavor profile compounds. The extracts were mainly composed of proteins (592 g kg-1 ) and minerals (386 g kg-1 ) and contained desirable flavor compounds. Their functional properties (solubility, water-holding capacity, oil-holding capacity) and antioxidant activities were also assessed, and their safety was verified. CONCLUSION: The cooking effluents generated by snow crab processing facilities, usually considered as waste, can be concentrated and turned into a natural aroma for the food industry. © 2019 Society of Chemical Industry.

Antihypertensive and Angiotensin-I-Converting Enzyme (ACE)-Inhibitory Peptides from Fish as Potential Cardioprotective Compounds.

The term metabolic/cardiometabolic/insulin resistance syndrome could generally be defined as the co-occurrence of several risk factors inclusive of systemic arterial hypertension. Not only that organizations, such as the world health organization (WHO) have identified high blood pressure as one of the main risk factors of the cardiometabolic syndrome, but there is also a link between the occurrence of insulin resistance/impaired glucose tolerance and hypertension that would consequently lead to type-2 diabetes (T2D). Hypertension is medicated by various classes of synthetic drugs; however, severe or mild adverse effects have been repeatedly reported. To avoid and reduce these adverse effects, natural alternatives, such as bioactive peptides derived from different sources have drawn the attention of researchers. Among all types of biologically active peptides inclusive of marine-derived ones, this paper's focus would solely be on fish and fishery by-processes' extracted peptides and products. Isolation and fractionation processes of these products alongside their structural, compositional and digestion stability characteristics have likewise been briefly discussed to better address the structure-activity relationship, expanding the reader's knowledge on research and discovery trend of fish antihypertensive biopeptides. Furthermore, drug-likeness of selected biopeptides was predicted by Lipinski's rules to differentiate a drug-like biopeptide from nondrug-like one.

Identification of A Novel Antibacterial Peptide from Atlantic Mackerel belonging to the GAPDH-Related Antimicrobial Family and Its In Vitro Digestibility.

The Atlantic mackerel, Scomber scombrus, is one of the most fished species in the world, but it is still largely used for low-value products, such as bait; mainly for crustacean fishery. This resource could be transformed into products of high value and may offer new opportunities for the discovery of bioactive molecules. Mackerel hydrolysate was investigated to discover antibacterial peptides with biotechnological potential. The proteolytic process generated a hydrolysate composed of 96% proteinaceous compounds with molecular weight lower than 7 kDa. From the whole hydrolysate, antibacterial activity was detected against both Gram-negative and Gram-positive bacteria. After solid phase extraction, purification of the active fraction led to the identification of 4 peptide sequences by mass spectrometry. The peptide sequence N-KVEIVAINDPFIDL-C, called Atlantic Mackerel GAPDH-related peptide (AMGAP), was selected for chemical synthesis to confirm the antibacterial activity and to evaluate its stability through in vitro digestibility. Minimal inhibitory concentrations of AMGAP revealed that Listeria strains were the most sensitive, suggesting potential as food-preservative to prevent bacterial growth. In addition, in vitro digestibility experiments found rapid (after 20 min) and early digestibility (stomach). This study highlights the biotechnological potential of mackerel hydrolysate due to the presence of the antibacterial AMGAP peptide.

Insights into the Regulation of Algal Proteins and Bioactive Peptides Using Proteomic and Transcriptomic Approaches.

Oceans abound in resources of various kinds for R&D and for commercial applications. Monitoring and bioprospecting allow the identification of an increasing number of key natural resources. Macroalgae are essential elements of marine ecosystems as well as a natural resource influenced by dynamic environmental factors. They are not only nutritionally attractive but have also demonstrated potential health benefits such as antioxidant, antihypertensive, and anti-inflammatory activities. Several bioactive peptides have been observed following enzymatic hydrolysis of macroalgal proteins. In addition, significant differences in protein bioactivities and peptide extracts of wild and cultivated macroalgae have been highlighted, but the metabolic pathways giving rise to these bioactive molecules remain largely elusive. Surprisingly, the biochemistry that underlies the environmental stress tolerance of macroalgae has not been well investigated and remains poorly understood. Proteomic and functional genomic approaches based on identifying precursor proteins and bioactive peptides of macroalgae through integrated multi-omics analysis can give insights into their regulation as influenced by abiotic factors. These strategies allow evaluating the proteomics profile of regulation of macroalgae in response to different growth conditions as well as establishing a comparative transcriptome profiling targeting structural protein-coding genes. Elucidation of biochemical pathways in macroalgae could provide an innovative means of enhancing the protein quality of edible macroalgae. This could be ultimately viewed as a powerful way to drive the development of a tailored production and extraction of high value molecules. This review provides an overview of algal proteins and bioactive peptide characterization using proteomics and transcriptomic analyses.

Identification of Peptides Implicated in Antibacterial Activity of Snow Crab Hepatopancreas Hydrolysates by a Bioassay-Guided Fractionation Approach Combined with Mass Spectrometry.

Snow crab (Chionoecetes opilio) by-products are a rich source of biomolecules, such as lipids, proteins, and chitin, which have not been extensively investigated. This study aims to identify antibacterial peptides to enhance the value of C. opilio by-products. After hydrolysis of different component parts using Protamex®, and concentration by solid-phase extraction, the resulting fractions were tested for antibacterial activity against Escherichia coli, Listeria innocua, and Vibrio parahaemolyticus. Hepatopancreas was the only tissue to display antibacterial activity detected using this protocol. Four fractions obtained with and without enzymatic hydrolysis of hepatopancreas followed by SPE C18 fractionation and elution with 50 and 80% acetonitrile demonstrated bacteriostatic activity against L. innocua HPB13, from concentrations of 0.30 to 43.05 mg/mL of peptides/proteins. Eleven peptides sharing at least 80% amino acid homology with four antimicrobial peptides were identified by mass spectrometry. Two peptides had homology to crustin-like and yellowfin tuna GAPDH antimicrobial peptides belonging to the marine organisms Penaeus monodon and Thunnus albacares, respectively. Other peptide sequence homologies were also identified: Odorranain-C7 from the frog Odorrana grahami and a predicted antibacterial peptide in the Asian ladybeetle Harmonia axyridis. These active peptides may represent a novel group of bioactive peptides deserving further investigation as food preservatives.

Silver nanoparticles and dissolved silver activate contrasting immune responses and stress-induced heat shock protein expression in sea urchin.

Using immune cells of sea urchin Strongylocentrotus droebachiensis in early development as a model, the cellular protective mechanisms against ionic and poly(allylamine)-coated silver nanoparticle (AgNPs; 14 ± 6 nm) treatments at 100 µg L-1 were investigated. Oxidative stress, heat shock protein expression, and pigment production by spherulocytes were determined as well as AgNP translocation pathways and their multiple effects on circulating coelomocytes. Sea urchins showed an increasing resilience to Ag over time because ionic Ag is accumulated in a steady way, although nanoAg levels dropped between 48 h and 96 h. A clotting reaction emerged on tissues injured by dissolved Ag (present as chloro-complexes in seawater) between 12 h and 48 h. Silver contamination and nutritional state influenced the production of reactive oxygen species. After passing through coelomic sinuses and gut, AgNPs were found in coelomocytes. Inside blood vessels, apoptosis-like processes appeared in coelomocytes highly contaminated by poly(allylamine)-coated AgNPs. Increasing levels of Ag accumulated by urchins once exposed to AgNPs pointed to a Trojan-horse mechanism operating over 12-d exposure. However, under short-term treatments, physical interactions of poly(allylamine)-coated AgNPs with cell structures might be, at some point, predominant and responsible for the highest levels of stress-related proteins detected. The present study is the first report detailing nano-translocation in a marine organism and multiple mechanisms by which sea urchin cells can deal with toxic AgNPs. Environ Toxicol Chem 2017;36:1872-1886. © 2016 SETAC.

Collagencin, an antibacterial peptide from fish collagen: Activity, structure and interaction dynamics with membrane.

In this study, we first report characterization of collagencin, an antimicrobial peptide identified from fish collagen hydrolysate. The peptide completely inhibited the growth of Staphylococcus aureus at 1.88 mM. Although non-toxic up to 470 µM, collagencin was hemolytic at higher concentrations. The secondary structure of collagencin was mainly composed by ß-sheet and ß-turn as determined by CD measurements and molecular dynamics. The peptide is likely to form ß-sheet structure under hydrophobic environments and interacts with both anionic (phosphatidylglycerol) and zwitterionic (phosphoethanolamine and phosphatidylcholine) lipids as shown with CD spectroscopy and molecular dynamics. The peptide formed several hydrogen bonds with both POPG and POPE lipids and remained at membrane-water interface, suggesting that collagencin antibacterial action follows a carpet mechanism. Collagenous fish wastes could be processed by enzymatic hydrolysis and transformed into products of high value having functional or biological properties. Marine collagens are a promising source of antimicrobial peptides with new implications in food safety and human health.

Purification and characterization of four antibacterial peptides from protamex hydrolysate of Atlantic mackerel (Scomber scombrus) by-products.

Proteins from fish by-product sources are valuable source of bioactive peptides and show promise as functional foods ingredients. The objective of the present study was to isolate and characterize antibacterial peptides from protamex hydrolysates of Atlantic mackerel (Scomber scombrus) by-products. Four sequences SIFIQRFTT (P4), RKSGDPLGR (P8.1), AKPGDGAGSGPR (P8.2) and GLPGPLGPAGPK (P11) were identified in peptide fractions separated using RP-HPLC. At 200 µg mL(-1), while peptides P8.1, P8.2 and P11 exhibited partial inhibition, P4 totally inhibited tested Gram-positive (Listeria innocua) and Gram-negative (Escherichia coli) bacterial strains. These results suggest that the protein hydrolysate derived from mackerel by-products could be used as an antimicrobial ingredient in both functional food and nutraceutical applications.

Substance misuse in the psychiatric emergency service; a descriptive study.

Substance misuse is frequently encountered in the psychiatric emergency service (PES) and may take many forms, ranging from formal DSM-IV diagnoses to less obvious entities such as hazardous consumption. Detecting such patients using traditional screening instruments has proved problematic. We therefore undertook this study to more fully characterize substance misuse in the PES and to determine whether certain variables might help better screen these patients. We used a prospectively acquired database of over 18,000 visits made to four PESs during a 2-year period in the province of Quebec, Canada. One of the variables acquired was a subjective rating by the nursing staff as to whether substance misuse was a contributing factor to the visit (graded as direct, indirect, or not at all). Substance misuse accounted for 21% of all diagnoses and alcohol was the most frequent substance used. Patients were divided into those with primary (PSM), comorbid (CSM) or no substance misuse (NSM). Depressive disorders were the most frequent primary diagnoses in CSM, whereas personality and substance misuse disorders were frequent secondary diagnoses in PSM. Although many variables significantly differentiated the three groups, few were sufficiently detailed to be used as potential screening tools. Those situations that did have sufficient details included those with a previous history of substance misuse, substance misuse within 48 hours of the visit, and visits graded by the nursing staff as being directly and/or indirectly related to substance misuse. Variables related to substance misuse itself were the primary predictors of PSM and, less significantly, CSM. The nursing staff rating, although promising, was obtained in less than 30% of all visits, rendering its practical use difficult to assess.

Evidence of anti-proliferative activities in blue mussel (Mytilus edulis) by-products.

Shellfish waste components contain significant levels of high quality protein and are therefore a potential source for biofunctional high-value peptides. The feasibility of applying a pilot scale enzymatic hydrolysis process to whole Mytilus edulis and, by fractionation, recover hydrolysates presenting a biological activity of interest, was evaluated. Fractions were tested on four immortalized cancerous cell lines: A549, BT549, HCT15 and PC3. The 50 kDa fraction, enriched in peptides, presented anti-proliferative activity with all cell lines and results suggest a bioactive molecule synergy within the fraction. At a protein concentration of 44 µg/mL, the 50 kDa fraction induced a mortality of 90% for PC3, 89% for A549, 85% for HCT15 and of 81% for BT549 cell lines. At the low protein concentration of only 11 µg/mL the 50 kDa fraction still entails a cell mortality of 76% for A549 and 87% for PC3 cell lines. The 50 kDa fraction contains 56% of proteins, 3% of lipids and 6% of minerals on a dry weight basis and the lowest levels detected of taurine and methionine and highest levels of threonine, proline and glycine amino acids. The enzymatic hydrolysis process suggests that Mytilus edulis by-products should be viewed as high-valued products with strong potential as anti-proliferative agent and promising active ingredients in functional foods.