Valorization of Yellowfin Tuna Tails: From Proteolytic Enzyme Production to Gelatin and Antioxidant Hydrolysate Extraction.

This study investigates the valorization potential of yellowfin tuna (Thunnus albacares) tails to produce high-value commercial products. Firstly, the tuna tails were placed in a perforated stainless-steel cylinder, and hydraulic pressure was applied to separate the skin from the muscle in the tails. The extracted muscle was then utilized as a nitrogen source for the growth of the proteolytic enzyme producer Bacillus subtilis, while the skins were employed for gelatin extraction. The proteases from B. subtilis were partially purified and used to produce antioxidant peptides from the obtained gelatin. The gelatin formed a gel upon cooling, with gelling and melting temperatures of 16 °C and 22 °C, respectively, and a Bloom strength of approximately 160. Response Surface Methodology (RSM) was employed to determine the optimal hydrolysis conditions to achieve the highest antioxidant activity (35.96% measured as DPPH radical scavenging activity), which were 50 °C and 6.5 IU of enzyme. The findings emphasize the importance of an integrated approach to maximize the value of tuna by-products, promoting sustainability within the framework of a circular bioeconomy. Overall, these results contribute to the efficient utilization of tuna by-products, waste reduction, and enhanced economic viability of the tuna industry.

Agro-Industrial Protein Waste and Co-Products Valorization for the Development of Bioplastics: Thermoprocessing and Characterization of Feather Keratin/Gliadin Blends.

Biopolymers based on plant and animal proteins are interesting alternatives in the development of films with future prospects as food packaging. Considering that in recent years there has been an increasing interest in the valorization of agro-industrial residues and by-products and that the blending of polymers can lead to materials with improved properties, in this work, keratin-rich feather fibers and gliadins were blended at different ratios in order to develop sustainable and biodegradable films. Control gliadin G100, feather F100 films, and their blends at 3:1 (G75F25), 2:2 (G50F50), and 1:3 (G25F75) ratios were successfully developed through thermoprocessing. The physical properties were differentiated as a function of the concentration of both polymeric matrices. Although gliadins showed higher hydrophilicity as confirmed by their highest swelling degree, films with high gliadin ratios exhibited lower water vapor permeability values at low and medium relative humidities. On the other hand, the feather fiber-based films displayed the highest Young's modulus values and provided an oxygen barrier to the blends, principally at the highest relative humidity. In conclusion, the blend of these protein-based polymers at different ratio resulted in interesting composites whose physical properties could be adjusted.

Evaluation of by-products from agricultural, livestock and fishing industries as nutrient source for the production of proteolytic enzymes.

This study presents an approach that utilizes low-value agro-industrial by-products as culture media for producing high-value proteolytic enzymes. The objective was to assess the impact of six agro-industrial by-products as culture media on the production of proteolytic enzymes. Bacillus subtilis strains, confirmed through comprehensive biochemical, morphological, and molecular analyses, were isolated and identified. Enzymatic activity was evaluated using azocasein and casein substrates, and the molecular sizes of the purified extract components were determined. The results demonstrated that the isolated bacteria exhibited higher metabolic and enzymatic activity when cultured in media containing 1 % soybean oil cake or feather meal. Furthermore, higher concentrations of the culture media were found to hinder the production of protease. Optimal protease synthesis on soybean oil cake and feather meal media was achieved after 4 days, using both the azocasein and casein methods. Semi-purification of the enzymatic extract obtained from Bacillus subtilis in feather meal and soybean oil cake resulted in a significant increase in azocaseinolytic and caseinolytic activities. Gel electrophoresis analysis revealed multiple bands in the fractions with the highest enzymatic activity in soybean oil cake, indicating the presence of various enzymes with varying molecular sizes. These findings highlight the potential of utilizing low-value agro-industrial by-products as efficient culture media for the sustainable and economically viable production of proteolytic enzymes with promising applications in various industries.

Influence of the Oil Structuring System on Lipid Hydrolysis and Bioaccessibility of Healthy Fatty Acids and Curcumin.

Oleogels (OG) and gelled emulsions (GE) were elaborated with a mixture of olive and chia oils (80:20 ratio) without and with the incorporation of the health-related compound curcumin. These were studied to evaluate the influence of the oil structuring system on the lipid hydrolysis and bioaccessibility of three healthy fatty acids (FA) (palmitic, oleic, and α-linolenic acids) and of curcumin, compared to the oil mixture (bulk oil, BO). The oil structuring system influenced the firmness and texture, and the presence of curcumin significantly altered the color parameters. GE showed higher lipid digestibility, with a greater proportion of absorbable fraction (higher content of free FA and monoacylglycerides) than OG, which behaved similarly to BO. The presence of curcumin affected the degree of lipolysis, reducing lipid digestibility in OG and increasing it in GE. As for FA bioaccessibility, although GE presented higher percentages overall, curcumin significantly increased and decreased FA bioaccessibility in OG and GE, respectively. The oil structuring system also influenced the bioaccessibility of curcumin, which was higher in GE. Therefore, when selecting an oil structuring system, their physicochemical properties, the degree of lipid hydrolysis, and the bioaccessibility of both curcumin and the FA studied should all be considered.

Protein Hydrolysis and Glycosylation as Strategies to Produce Bioactive Ingredients from Unmarketable Prawns.

The present work shows a procedure to valorize non-commercial boiled shrimp to produce functional ingredients, using a combined treatment based on enzymatic hydrolysis and subsequent glycation under mild conditions. Antioxidant and prolyl endopeptidase-inhibiting activities were determined as a function of hydrolysis and glycation times (0-120 min and 0-180 min, respectively). The reaction products were characterized by determining the degree of hydrolysis, browning, fluorescent compounds, free amino acids, phenol content, Fourier transform infrared spectroscopy (FTIR), and molecular weight of the different fractions obtained. Enzymatic hydrolysis generated hydrolysates with significant antioxidant and prolyl endopeptidase-inhibiting activities. Glycation under mild conditions was used as a strategy to improve the antioxidant and potential nootropic properties of the hydrolysates. During glycation, the free amino acid content decreased, total phenols and fluorescent compounds increased significantly, and low molecular weight melanoidins were formed. The presence of peptide-glucose conjugates was also confirmed by FTIR. Glycation increased the antioxidant activities of the hydrolysates; however, their prolyl-endopeptidase-inhibiting activity was lost. Results showed that compounds with promising antioxidant (hydrolysis and glycation) and potential nootropic (hydrolysis) activities and applications in food systems were obtained from the biotechnological strategy used.

The Effect of Emulsifying Protein and Addition of Condensed Tannins on n-3 PUFA Enriched Emulsions for Functional Foods.

This paper examines the effect of the type of the emulsifying protein (EP) (sodium caseinate (SC) and whey protein isolate (WPI)) on both oil-in-water liquid-like emulsions (Es) and the corresponding cold gelled emulsions (GEs), and also the effect of addition of carob extract rich in condensed tannins (T). The systems, intended as functional food ingredients, were studied in various different respects, including rheological behaviour, in vitro gastrointestinal digestion with determination of the release of non-extractable proanthocyanidins (NEPA) from T, antioxidant activity and lipolysis. EP significantly affects the rheological behaviour of both Es and GEs. T incorporation produced a structural reinforcement of GEs, especially in the case of SC. The digests from Es displayed a higher antioxidant activity than those from GEs. T lipase inhibition was observed only in the formulations with WPI. Our results highlight the importance, in the design of functional foods, of analyzing different variables when incorporating a bioactive compound into a food or emulsion in order to select the better combination for the desired objective, owing to the complex interplay of the various components.

Recent Advances in Astaxanthin Micro/Nanoencapsulation to Improve Its Stability and Functionality as a Food Ingredient.

Astaxanthin is a carotenoid produced by different organisms and microorganisms such as microalgae, bacteria, yeasts, protists, and plants, and it is also accumulated in aquatic animals such as fish and crustaceans. Astaxanthin and astaxanthin-containing lipid extracts obtained from these sources present an intense red color and a remarkable antioxidant activity, providing great potential to be employed as food ingredients with both technological and bioactive functions. However, their use is hindered by: their instability in the presence of high temperatures, acidic pH, oxygen or light; their low water solubility, bioaccessibility and bioavailability; their intense odor/flavor. The present paper reviews recent advances in the micro/nanoencapsulation of astaxanthin and astaxanthin-containing lipid extracts, developed to improve their stability, bioactivity and technological functionality for use as food ingredients. The use of diverse micro/nanoencapsulation techniques using wall materials of a different nature to improve water solubility and dispersibility in foods, masking undesirable odor and flavor, is firstly discussed, followed by a discussion of the importance of the encapsulation to retard astaxanthin release, protecting it from degradation in the gastrointestinal tract. The nanoencapsulation of astaxanthin to improve its bioaccessibility, bioavailability and bioactivity is further reviewed. Finally, the main limitations and future trends on the topic are discussed.

Functional aptitude of hake minces with added TMAO-demethylase inhibitors during frozen storage.

The ability of compounds of natural origin (black, white, red, and green tea extracts, phytic acid) to inhibit TMAO-demethylase enzyme was assayed. Black tea and phytic acid exerted the highest inhibiting activities, similar to the already known inhibitor sodium citrate. Hake minces incorporating these three compounds were prepared and stored frozen (150 days, -12 °C). TMAO-demethylase enzyme was partially inhibited (lower enzyme activity, reduction of formaldehyde accumulation). The study of physicochemical properties of the minces (salt-soluble proteins, water holding capacity, structural water associated with myofibrils) pointed to evident protein aggregation and loss of functionality when phytic acid was added, whereas black tea and sodium citrate did not have a negative effect. Consequently, the salt-ground mince with phytic acid showed worse viscoelastic properties than the others. In conclusion, black tea polyphenols and sodium citrate can be used as additives to inhibit TMAO-demethylase enzyme during frozen storage of fish minces.

Bioaccessibility and antimicrobial properties of a shrimp demineralization extract blended with chitosan as wrapping material in ready-to-eat raw salmon.

A shrimp extract (SME) obtained from the mild-acid demineralization treatment of shrimp shells to produce chitosan was collected. It was mainly composed of fat (≈73%), protein (≈19%), and ash (≈9%) and contained considerable amounts of calcium (≈1.9 g/100 g), astaxanthin (≈30 mg/100 g) and unsaturated fatty acids (≈27% MUFA, ≈39% PUFA). The SME was used in combination with chitosan for wrapping raw salmon to produce a ready-to-eat product enriched in calcium. No significant changes in hardness were found, as compared to the unwrapped salmon. Estimated intakes of bioaccessible calcium increased significantly by 3.6-fold, whereas intake of bioaccessible fat was reduced by 15%. SFA were the main fatty acid group reduced (≈80%), whereas MUFA and PUFA were only reduced by ≈20% each. Total viable counts, pseudomonads, enterobacteria, and specific fish spoilers were reduced by 2-4 log CFU/g in wrapped sample during the chilled storage period (19 days).

Emulsion gels containing n-3 fatty acids and condensed tannins designed as functional fat replacers.

The purpose of this study was to ascertain the potential of several food-grade emulsion (O/W) gels (GEs) for use as healthier fat replacers. The emulsions, formulated with a lipid phase rich in n-3 fatty acids and different emulsifiers (sodium caseinate, SC; whey protein isolate, WPI and isolated soy protein, ISP), were cold gelled after adding a natural extract rich in condensed tannins (CT). The GEs were characterized and their oxidative stability evaluated during storage (4 °C). All GEs formulated presented a solid-like structure showing generally excellent emulsion stability, which improved in GEs with the addition of CT. Non-extractable proanthocyanidins (NEPA) were the main source of polyphenol in samples enriched with CTs. The antioxidant activity of the systems was not affected by the use of different proteins as emulsifiers, but it was improved in GEs containing CT. The oxidation values recorded in the GE systems can generally be regarded as low even considering their enrichment with unsaturated fatty acids, which thus assures their suitability for use as fat replacers.

Chemical characterization of wash water biomass from shrimp surimi processing and its application to develop functional edible films.

The aim of this work was to recover and study the composition of the biomass obtained from the surimi processing industry, and to explore the feasibility of its valorization by developing functional edible films. A concentrate from wash water of minced shrimp muscle of low commercial value [shrimp concentrate (SC)] was obtained by alkaline solubilization and isoelectric precipitation, which had protein content of 61.8% and fat 23.6%. The protein fraction of low molecular weight (≤37 kDa), with maximum solubility at basic pH 9-11, and was rich in essential amino acids (296 essential amino acids/1000 residues), whereas the fat fraction showed a high degree of unsaturation (26.8% MUFA, 28.3% PUFA). SC, showed antioxidant activity measured by FRAP and ABTS assays and, was successfully incorporated into an agar film matrix (agar-to-protein ratio of 3:1). The films were translucent and brownish and maintained integrity upon immersion in water, but swelled considerably. Films were able to release protein and exert antioxidant activity in water and 95% ethanol (aqueous and fatty food simulants, respectively), the release being faster and greater in the former. In conclusion, the biomass obtained from shrimp mince/surimi processing was for developing edible films.

The effect of the combined use of high pressure treatment and antimicrobial edible film on the quality of salmon carpaccio.

Fish carpaccio is a ready-to-eat product with a very limited shelf life. In the present work, the use of high pressure treatment (HP) and/or antimicrobial edible film was studied in order to improve quality and stability of salmon carpaccio. In a preliminary part of the work, a film composed of gelatin plus chitosan incorporating clove essential oil was selected, based on its physicochemical and antimicrobial properties. Eugenol and ß-caryophyllene, the main volatile components of the film, migrated to salmon muscle, the release being favored by HP and storage time. Concurrently, reducing power of the muscle increased, resulting in prevention of lipid oxidation derived from either HP or refrigerated storage. HP treatment reduced total microbial counts by 1.5 log cycles from the onset of storage, whereas the film reduced it by 2 log cycles after 3 days. The combination of HP and edible film exerted the most intense antimicrobial effect, total bacterial counts, luminescent bacteria, H2S-producing organisms, pseudomonads, enterobacteria, and lactic acid bacteria remaining constant or under detection limit over the whole storage period (11 days). The combined use of HP treatment and gelatin-chitosan-clove essential oil film is an effective way of improving quality and stability of salmon carpaccio.

Effect of different polysaccharides and crosslinkers on echium oil microcapsules.

Microencapsulation by complex coacervation using gelatin and arabic gum (AG) as wall materials and transglutaminase for crosslinking is commonly used. However, AG is only produced in a few countries and transglutaminase is expensive. This work aimed to evaluate the encapsulation of echium oil by complex coacervation using gelatin and cashew gum (CG) as wall materials and sinapic acid (S) as crosslinker. Treatments were analyzed in relation to morphology, particle size, circularity, accelerated oxidation and submitted to different stress conditions. Rounded microcapsules were obtained for treatments with AG (45.45µm) and microcapsules of undefined format were obtained for treatments with CG (22.06µm). The S incorporation for 12h improved the oil stability by three fold compared to oil encapsulated without crosslinkers. Treatments with CG and S were resistant to different stress conditions similar to treatments with AG and transglutaminase, making this an alternative for delivery/application of compounds in food products.

Functional properties and antifungal activity of films based on gliadins containing cinnamaldehyde and natamycin.

Gliadin films cross-linked with cinnamaldehyde (1.5, 3, and 5%) and incorporated with natamycin (0.5%) were prepared by casting, and their antifungal activity, water resistance, and barrier properties were characterized. Incorporation of natamycin gave rise to films with greater water uptake, weight loss and diameter gain, and higher water vapor and oxygen permeabilities. These results may be associated to a looser packing of the protein chains as a consequence of the presence of natamycin. The different cross-linking degree of the matrices influenced the natamycin migration to the agar test media, increasing from 13.3 to 23.7 (µg/g of film) as the percentage of cinnamaldehyde was reduced from 5% to 1.5%. Antifungal activity of films was assayed against common food spoilage fungi (Penicillium species, Alternaria solani, Colletotrichum acutatum). The greatest effectiveness was obtained for films containing natamycin and treated with 5% of cinnamaldehyde. The level of cinnamaldehyde reached in the head-space of the test assay showed a diminishing trend as a function of time, which was in agreement with fungal growth and cinnamaldehyde metabolization. Developed active films were used in the packaging of cheese slices showing promising results for their application in active packaging against food spoilage.

The effect of combined traditional and novel treatments on oxidative status of dolphinfish (Coryphaena hippurus) and sardine (Sardina pilchardus) muscle lipids.

Fish is rich in polyunsaturated fatty acids with beneficial effects on human health; however, these lipids are very sensitive to auto-oxidation reactions, leading to loss of nutritional and sensory quality. The effect of traditional (brining, smoking) and novel (addition of polyphenolic extracts, high pressure) preservation processes on the antioxidant/oxidative status of muscle lipids from dolphinfish and sardine was studied. Brining with oregano or rosemary aqueous extracts, as well as smoking, gave rise to deposition of phenolic compounds (9-42, 1.5-4.5 and 0.4-2.3 µg phenol/g for smoked, oregano-brined and rosemary-brined samples, respectively) in the muscle of both fish species. The antioxidant activity, as measured by ferric reducing ability, was also improved after brining with antioxidant extracts or smoking, results ranging from 8.9 to 82 mM FeSO4 · 7H2O equivalents/mg muscle depending on the treatment and the fish species. Consequently, fish lipid oxidation (as measured by thiobarbituric acid reactive substances) derived from brining and pressurizing, applied alone or in combination, was reduced between 6.6 and 69.8% depending on the treatment and the fish species. The combination of brining with oregano extract and light smoking showed an additional antioxidant effect, as compared with that obtained by smoking, on reducing sardine lipid oxidation derived from brining and pressurizing.

Biochemical properties of bioplastics made from wheat gliadins cross-linked with cinnamaldehyde.

The aim of this work has been to study the modification of gliadin films with cinnamaldehyde as a potential cross-linker agent. The molecular weight profile and cross-linking density showed that cinnamaldehyde increased reticulation in the resulting films. The participation of free amino groups of the protein in the newly created entanglements could be a possible mechanism of connection between the polypeptidic chains. The combination of a Schiff base and a Michael addition is a feasible approach to understanding this mechanism. The protein solubility in different media pointed to lower participation by both noncovalent and disulfide bonds in stabilizing the structure of the cross-linked films. The new covalent bonds formed by the cinnamaldehyde treatment hampered water absorption and weight loss, leading to more water-resistant matrices which had not disintegrated after 5 months. The properties of this novel bioplastic could be modified to suit the intended application by using cinnamaldehyde, a naturally occurring compound.

Functional properties of bioplastics made from wheat gliadins modified with cinnamaldehyde.

Cinnamaldehyde is a naturally occurring α,ß-unsaturated aldehyde. Its potential as a natural cross-linker to improve the physical performance of cast wheat gliadin films was evaluated. The cross-linking reaction was found to be dependent on the pH of the reaction medium, with pH 2 as the optimum. The water resistance (weight loss after immersion), mechanical properties (Young's modulus, tensile strength and elongation at break), thermal properties (T(g) and decomposition behavior), optical properties and morphology of films were evaluated. Cross-linked films showed high transparency, maintained their integrity after immersion, and displayed significant improvements in tensile strength and Young's modulus without impairment of their elongation properties. These effects, which were proportional to the amount of cinnamaldehyde added, highlight the possible formation of intermolecular covalent bonds between "monomeric" gliadins, leading to a polymerized network. Thus, this treatment could provide a new alternative to the toxic cross-linkers commonly employed and so extend the use of gliadin films.