Algae-derived compounds: Bioactivity, allergenicity and technologies enhancing their values.

As a rapidly growing source of human nutrients, algae biosynthesize diverse metabolites which have promising bioactivities. However, the potential allergenicity of algal components hinder their widespread adoption. This review provides a comprehensive review of various macro and micronutrients derived from algal biomass, with particular focus on bioactive compounds, including peptides, polyphenols, carotenoids, omega-3 fatty acids and phycocyanins. The approaches used to produce algal bioactive compounds and their health benefits (antioxidant, antidiabetic, cardioprotective, anti-inflammatory and immunomodulatory) are summarised. This review particularly focuses on the state-of-the-art of precision fermentation, encapsulation, cold plasma, high-pressure processing, pulsed electric field, and subcritical water to reduce the allergenicity of algal compounds while increasing their bioactivity and bioavailability. By providing insights into current challenges of algae-derived compounds and opportunities for advancement, this review contributes to the ongoing discourse on maximizing their application potential in the food nutraceuticals, and pharmaceuticals industries.

A review on proteomic and genomic biomarkers for gelatin source authentication: Challenges and future outlook.

Biomarkers are compounds that could be detected and used as indicators of normal and/or abnormal functioning of different biological systems, including animal tissues and food matrices. Gelatin products of animal origin, mainly bovine and porcine, are currently under scrutiny mainly due to the specific needs of some sectors of the population related to religious beliefs and their dietary prohibitions, as well as some potential health threats associated with these products. Thus, manufacturers are currently in need of a reliable, convenient, and easy procedure to discern and authenticate the origin of animal-based gelatins (bovine, porcine, chicken, or fish). This work aims to review current advances in the creation of reliable gelatin biomarkers for food authentication purposes based on proteomic and DNA biomarkers that could be applied in the food sector. Overall, the presence of specific proteins and peptides in gelatin can be chemically analysed (i.e., by chromatography, mass spectroscopy, electrophoresis, lateral flow devices, and enzyme-linked immunosorbent assay), and different polymerase chain reaction (PCR) methods have been applied for the detection of nucleic acid substances in gelatin. Altogether, despite the fact that numerous methods are currently being developed for the purpose of detecting gelatin biomarkers, their widespread application is highly dependent on the cost of the equipment and reagents as well as the ease of use of the various methods. Combining different methods and approaches targeting multiple biomarkers may be key for manufacturers to achieve reliable authentication of gelatin's origin.

Strategies for oral delivery of bioactive peptides with focus on debittering and masking.

Protein hydrolysis is a process used in the food industry to generate bioactive peptides of low molecular weight and with additional health benefits, such as antihypertensive, antidiabetic, and antioxidant properties that are often associated with their content on hydrophobic amino acids. This results in an increased bitterness of the products, making them less desirable for their use in food formulations. This review summarizes the main dietary sources of bitter bioactive peptides, including methods to determine their bitterness, such as the Q-values and electronic tongue; and the main factors and mechanisms underlying the bitterness of these compounds. The main strategies currently used to improve the taste and oral delivery of bioactive peptides are also discussed together with the main advantages and drawbacks of each technique. Debittering and masking techniques are reported in detail, including active carbon treatments, alcohol extraction, isoelectric precipitation, chromatographic methods, and additional hydrolytic processes. Other masking or blocking techniques, including the use of inhibitors, such as modified starch, taurine, glycine, and polyphosphates, as well as chemical modifications, such as amination, deamination, acetylation, or cross-linking were also discussed. The findings of this work highlight encapsulation as a highly effective method for masking the bitter taste and promoting the bioactivity of peptides compared to other traditional debittering and masking processes. In conclusion, the article suggests that advanced encapsulation technologies can serve as an effective means to mitigate the bitterness associated with bioactive peptides, while simultaneously preserving their biological activity, increasing their viability in the development of functional foods and pharmaceuticals.

Advanced Processing of Giant Kelp (Macrocystis pyrifera) for Protein Extraction and Generation of Hydrolysates with Anti-Hypertensive and Antioxidant Activities In Vitro and the Thermal/Ionic Stability of These Compounds.

In this study, giant kelp was explored under various conventional and ultrasound-assisted extraction (UAE) conditions for the extraction of protein, its hydrolysis, and ultrafiltration to generate multiple fractions. The amino acid composition of all the fractions and their biological activities in vitro, including angiotensin-converting enzyme I (ACE) inhibitory activity and antioxidant activities (2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, reducing power (RP), and ferrous chelating (FC) activities) were tested by storing the compounds for 2 weeks at various temperatures (-20-60 °C) and pHs (2-11) to elucidate their thermal and ionic stability, respectively. The yield of protein extraction using the conventional method was lower (≈39%) compared to the use of UAE (150 W, 15 min), which achieved protein recoveries of approximately 60%. After enzymatic hydrolysis and ultrafiltration, low-molecular-weight (MW) hydrolysates had the highest levels of ACE inhibitory (80%), DPPH (84%), RP (0.71 mM trolox equivalents), and FC (81%) activities. Amino acids associated with peptides of high biological activities, such as Val, Ala, Asx, Gly, Lys, Met, Leu, and His, were at higher levels in the low MW fraction compared to any other sample. The biological activities in vitro of all the samples fluctuated under the multiple storage conditions studied, with the highest stability of all the samples appreciated at -20 °C and pH 7. This study shows for the first time the use of giant kelp as a promising source of bioactive peptides and indicates the optimum processing and storing conditions for the use of these compounds as nutraceuticals or functional foods that could help in the prevention of cardiovascular disorders and multiple chronic diseases associated with oxidative damage.

Influence of Enzymatic Hydrolysis and Molecular Weight Fractionation on the Antioxidant and Lipase / α-Amylase Inhibitory Activities In Vitro of Watermelon Seed Protein Hydrolysates.

This study aims to evaluate the potential in vitro antioxidant and anti-obesity activities of watermelon seed protein hydrolysates (WSPH) obtained using different combinations of enzymes alcalase−proteinase K (ALC-PK) and alcalase−actinidin (ALC-ACT). There was a direct relationship between the degree of hydrolysis (DH) and the biological activities of the WSPH, with the highest DPPH (approximately 85%) and lipase inhibitory activities (≈59%) appreciated at DH of 36−37% and 33−35% when using ALC-PK and ALC-ACT, respectively. Following molecular weight fractionation, the ALC-PK WSPH < 3 kDa (F1) assayed at 1 mg.mL−1 had the highest DPPH-radical scavenging (89.22%), ferrous chelating (FC) (79.83%), reducing power (RP) (A 0.51), lipase inhibitory (71.36%), and α-amylase inhibitory (62.08%) activities. The amino acid analysis of ALC-PK WSPH and its fractions revealed a relationship between the biological activity of the extracts and their composition. High contents of hydrophobic amino acids, arginine, and aromatic amino acids were related to high antioxidant, lipase inhibitory, and α-amylase inhibitory activities in the extracts, respectively. Overall, this study revealed that underutilized protein sources such as WSPH, using the appropriate combination of enzymes, could result in the generation of new ingredients and compounds with powerful antioxidant and anti-obesity activities with promising applications as nutraceuticals or functional foods.

Impact of Whey Protein Edible Coating Containing Fish Gelatin Hydrolysates on Physicochemical, Microbial, and Sensory Properties of Chicken Breast Fillets.

This study aims to research the impact of coatings containing whey protein (WP), fish gelatin hydrolysates (FGH), and both compounds together (WP + FGH) on the shelf-life of chicken breast fillets over the course of 16 days of cold storage (4 °C, 4-day intervals), as assessed by their physicochemical, microbiological, and sensory properties. Overall, cooking loss, pH value, total volatile base nitrogen, free fatty acids, peroxide value, and thiobarbituric acid reactive substances increased with storage time in all samples. WP + FGH coated samples had significantly lower variation in all these parameters over the time of storage compared to other coated samples (WP and FGH), while these parameters increased greatly in control (uncoated) samples. WP + FGH coating also resulted in reduced bacterial counts of total mesophilic, aerobic psychrotrophic, and lactic acid bacteria compared to other coated and uncoated samples. The sensory evaluation revealed no differences in the panelists' overall acceptance at day 0 of storage between samples. The samples were considered "non-acceptable" by day 8 of storage; however, WP + FGH coated samples maintained an overall higher acceptability score for the sensory attributes evaluated by the panelists. Overall, this study shows the potential of WP + FGH coatings for prolonging the shelf-life of chicken breast fillets.

Cardioprotective Peptides from Milk Processing and Dairy Products: From Bioactivity to Final Products including Commercialization and Legislation.

Recent research has revealed the potential of peptides derived from dairy products preventing cardiovascular disorders, one of the main causes of death worldwide. This review provides an overview of the main cardioprotective effects (assayed in vitro, in vivo, and ex vivo) of bioactive peptides derived from different dairy processing methods (fermentation and enzymatic hydrolysis) and dairy products (yogurt, cheese, and kefir), as well as the beneficial or detrimental effects of the process of gastrointestinal digestion following oral consumption on the biological activities of dairy-derived peptides. The main literature available on the structure-function relationship of dairy bioactive peptides, such as molecular docking and quantitative structure-activity relationships, and their allergenicity and toxicity will also be covered together with the main legislative frameworks governing the commercialization of these compounds. The current products and companies currently commercializing their products as a source of bioactive peptides will also be summarized, emphasizing the main challenges and opportunities for the industrial exploitation of dairy bioactive peptides in the market of functional food and nutraceuticals.

Wheat Germ Fermentation with Saccharomyces cerevisiae and Lactobacillus plantarum: Process Optimization for Enhanced Composition and Antioxidant Properties In Vitro.

Wheat germ, a by-product of the flour milling industry, is currently commercialized mainly for animal feed applications. This study aims to explore and optimize the process of wheat germ fermentation to achieve products with enhanced nutritional composition and biological properties and further characterize the fermented products generated using these optimum conditions. The type of microorganism (Saccharomyces cerevisiae 5022 (yeast) and Lactobacillus plantarum strain 299v (bacteria)), pH (4.5, 6, and 7.5) and fermentation time (24, 48, and 72 h) were optimized using response surface methodology (RSM) aiming to achieve fermented products with high total phenol content (TPC), dimethoxy benzoquinone (DMBQ) and antioxidant activities. Optimum fermentation conditions were achieved using L. plantarum, pH 6, 48 h, generating extracts containing TPC (3.33 mg gallic acid equivalents/g), DMBQ (0.56 mg DMBQ/g), and DPPH radical scavenging (86.49%). These optimally fermented products had higher peptide concentrations (607 µg/mL), gamma-aminobutyric acid (GABA) (19,983.88 mg/kg) contents compared to non-fermented or yeast-fermented products. These findings highlight the influence of fermentation conditions of wheat germ and the promising industrial application of wheat germ fermentation for developing food products with enhanced biological properties promising for their commercialization as functional foods.

Development of Cobalt-Binding Peptide Chelate from Human Serum Albumin: Cobalt-Binding Properties and Stability.

Radioactive isotopes are used as drugs or contrast agents in the medical field after being conjugated with chelates such as DOTA, NOTA, DTPA, TETA, CyDTA, TRITA, and DPDP. The N-terminal sequence of human serum albumin (HSA) is known as a metal binding site, such as for Co2+, Cu2+, and Ni2+. For this study, we designed and synthesized wAlb12 peptide from the N-terminal region of HSA, which can bind to cobalt, to develop a peptide-based chelate. The wAlb12 with a random coil structure tightly binds to the Co(II) ion. Moreover, the binding property of wAlb12 toward Co(II) was confirmed using various spectroscopic experiments. To identify the binding site of wAlb12, the analogs were synthesized by alanine scanning mutagenesis. Among them, H3A and Ac-wAlb12 did not bind to Co(II). The analysis of the binding regions confirmed that the His3 and α-amino group of the N-terminal region are important for Co(II) binding. The wAlb12 bound to Co(II) with Kd of 75 µM determined by isothermal titration calorimetry when analyzed by a single-site binding model. For the use of wAlb12 as a chelate in humans, its cytotoxicity and stability were investigated. Trypsin stability showed that the wAlb12 - Co(II) complex was more stable than wAlb12 alone. Furthermore, the cell viability analysis showed wAlb12 and wAlb12 + Co(II) to be non-toxic to the Raw 264.7 and HEK 293T cell lines. Therefore, a hot radioactive isotope such as cobalt-57 will have the same effect as a stable isotope cobalt. Accordingly, we expect wAlb12 to be used as a peptide chelate that binds with radioactive isotopes.

Structure-function relationship of fermented skate skin gelatin-derived bioactive peptides: a peptidomics approach.

In this study, we investigated the multi-functionality of bioactive peptides derived from fermented skate (Raja kenojei) skin gelatin hydrolysates. The extracted gelatin was hydrolyzed using a combination of food grade subtilisin and actinidin. The hydrolysates were then fractionated via ultrafiltration, and the fractions with the highest dipeptidyl peptidase-IV (DPP-IV) inhibitory, angiotensin-converting enzyme (ACE) inhibitory, and antibacterial proprieties were further purified via ion exchange, solid phase extraction, and reverse phase high performance liquid chromatography. Analysis of the obtained extract revealed a direct relationship between hydrolysis time, degree of hydrolysis, and biological activities. The peptides GRPGNRGE (P1) and AKDYEVDAT (P2), with a molecular weight of 841.42 and 1010.46 Da, respectively, were identified through tandem mass spectrometry. P1 had a lower ACE and DPP-IV inhibitory activity, with a half maximal inhibitory concentration [IC50] of 0.74 and 0.69 mg.mL-1, respectively, than P2 (0.52 and 0.58 mg.mL-1, respectively). Antibacterial analysis showed similar results, with a minimum inhibitory concentration of 0.52 and 0.46 mg.mL-1 against Staphylococcus aureus (highest activity) and 1.75 and 1.44 mg.mL-1 against Klebsiella pneumonia (lowest activity) for P1 and P2, respectively. Overall, this study revealed two fish gelatin-derived multifunctional peptides, exhibiting ACE inhibitory, DPP-IV inhibitory, and antibacterial activities, as natural nutraceuticals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00998-6.

Structure-function engineering of novel fish gelatin-derived multifunctional peptides using high-resolution peptidomics and bioinformatics.

The multifunctional properties of fish gelatin hydrolysates have not been completely elucidated. Here, the biological characterization of these peptides was performed to engineer multifunctional peptides. Bioactive peptides were produced from mackerel byproducts via successive enzymatic hydrolysis reactions using subtilisin A and actinidin as microbial and herbal proteases. The antibacterial activity against both gram-negative and -positive food-borne pathogens, including Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae, as well as the inhibitory potential of angiotensin-converting enzyme (ACE) and dipeptidyl peptidase IV (DPP-IV), was accessed in vitro. The synthesized peptides demonstrated multifunctional properties, which were further confirmed by in silico protocols. The ACE and DPP-IV inhibitory (IC50) values of P1, P2, and P3 were 0.92 and 0.87, 0.51 and 0.93, 0.78 and 1.16 mg mL-1, respectively. Moreover, the binding energy was sufficient for all three peptides to inhibit both ACE and DPP-IV enzymes with excellent three-dimensional conformation (RMSD = 0.000) for all six docking mechanisms.

Antioxidant mechanism, antibacterial activity, and functional characterization of peptide fractions obtained from barred mackerel gelatin with a focus on application in carbonated beverages.

The present study aimed to use fish by-products to generate gelatin peptides with potential applications in carbonated beverages. After ultrafiltration, the F < 3 kDa (fraction < 3 kDa) showed the highest peptide concentration (227.22 mg/g) as well as antibacterial (MIC of ≤ 0.5 mg/mL) and antioxidant activities, including hydroxyl and superoxide radical scavenging, ferrous chelation, and ferric reduction (with IC50 values of 0.88, 1.04, 0.50 mg/mL, and 0.58, respectively). 2,2-diphenyl-1-picrylhydrazyl scavenging was the highest in the 3 < F < 10 kDa (IC50 of 0.64 mg/mL). In vitro gastrointestinal digestion significantly decreased all biological activities. Solubility, water holding capacity, and emulsifying activity of the F < 3 kDa were the highest while foaming properties and overfoaming were reversibly related to the molecular weight. All abovementioned properties, in addition to in vitro cytotoxicity analysis in different cell lines and better flavor characteristics, indicated that the F < 3 kDa could be safely and properly used as an ingredient for the fortification of carbonated beverages.

The effects of enzymatically aided acid-swelling process on gelatin extracted from fish by-products.

The objective of this study was to investigate the effects of the enzymatic aided acid-swelling process on gelatin obtained from fish by-products. For this purpose, gelatin was extracted by an acidic swelling procedure in the presence of protease extracted from Rainbow trout pyloric caeca. The yield of gelatin extraction and the most important physicochemical characteristics of the fish gelatin samples were investigated and compared with those of commercial bovine gelatin (CBG). The yields of gelatin from Epinephelus coioides skin (ESG) either with or without crude protease from pyloric caeca (15 units/g alkaline treated) were 14.98% and 50.89%, respectively. The yields of gelatin from Cyprinus carpio scales (CSG) with crude protease from pyloric caeca (15 units/g) were 49.97%. The gel strength of the CSG (259.66 g) was significantly higher than that of CBG (228.30 g) and ESG (187.75 g). Similarly, the gelling and melting points, foaming capacity and stability, and the SDS-PAGE pattern of gelatins were compared. The electrophoretic pattern confirmed the results of gel strength which was due to the narrower alpha and beta bands in fish skin and commercial bovine gelatins than that of fish scales gelatin. The results of this research showed that the production of high-quality gelatin can be achieved by the enzymatically aided acid-swelling procedure from fish scales and skin.

Shelf-life extension of whole shrimp using an active coating containing fish skin gelatin hydrolysates produced by a natural protease.

This study was focused on shelf-life extension of whole shrimp (Penaeus merguiensis) using an active coating containing gelatin hydrolysates. Gelatin extracted from Scomberomorus commerson skin was hydrolyzed using actinidin extracted from kiwifruit. Some important physicochemical characteristics of fish skin gelatin including viscosity, gelling and melting points, and temperatures were examined. The whole shrimp was coated with four coating agents including fish skin gelatin (FG), commercial gelatin (CG), fish skin gelatin containing 1 mg/ml fish gelatin hydrolysates (FG + GH), and commercial bovine gelatin containing 1 mg/ml fish gelatin hydrolysates (CG + GH). Chemical, microbial, and sensorial properties of samples were monitored for 12 days at 4°C with 3-day intervals (0-12 days). The pH value of samples coated with FG + GH and CG + GH showed the lowest changes during 12 days of storage (1.68 ± 0.00 and 1.70 ± 0.09, respectively). The free fatty acid content (FFA), total volatile base nitrogen (TVB-N), lipid oxidation, and carbonyl content of samples coated with FG + GH and CG + GH were significantly lower than that of control, CG, and FG samples. The results of this study showed that the gelatin hydrolysates could be used as a preservative costing agent for whole shrimp.

Physicochemical, rheological, and molecular characterization of colloidal gelatin produced from Common carp by-products using microwave and ultrasound-assisted extraction.

The effects of ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) methods on molecular and physicochemical characteristics of the resultant gelatin were examined. Before extraction procedure, we investigated the optimum pH for swelling of Common carp by-products, which is an important pretreatment for gelatin production. The highest swelling yield was achieved at pH 13 among pH 1-14 with unit intervals. Results indicated that the UAE gelatin has a higher gel strength, viscosity, melting point, and gelling point. The power and time of sonication showed a reverse relation with these characteristics. In addition, as the time of microwave heating was raised, the gel strength, viscosity, melting point, and gelling point were decreased. The FT-IR spectra showed similar peaks but the Amide B in UAE gelatin slightly vanished. The electrophoretic pattern also revealed the higher gel strength and viscosity of UAE gelatin due to the higher intensity of α and ß chains compared to MAE gelatin. It can be concluded from all of the results of this study that the produced gelatin using these procedures can be a good source of gelatin in food and drug industries.

Gelatin production using fish wastes by extracted alkaline protease from Bacillus licheniformis.

An effort was made to produce gelatin from Common carp wastes using extracted alkaline protease from Bacillus licheniformis PTCC 1595. Fermentation was performed by submerged media for 48 h and 72 h. The hydrolyzing enzyme was added in 5, 10, 15, 20, and 25 units per gram of wastes powder for hydrolysis. The produced gelatin was compared with commercial bovine gelatin with regard to some rheological and physicochemical properties. The yield of gelatin production was also determined as a result of hydroxyproline extraction from fish wastes. SDS-PAGE was performed for enzyme and gelatins. For enzyme, two bands were achieved with 39 and 10.5 kDa molecular weight which were separated passing through a 15 kDa UF filter. Both gelatins showed ß, α1, and α2 chains as basic components, but the fish waste gelatin showed narrow bands. In conclusion, foam expansion and water holding capacity were approximately equal in both gelatin types used for food industry application. The results indicated that using 20 units of enzyme per gram of waste powder was the optimum amount of enzyme application. Further, fish wastes were concluded to be a practical source for gelatin production.