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The effects of cross-processing lingonberry press cake (LPC) (2.5-30 %, dw/dw) with herring co-products on protein yield, oxidative stability and color of pH-shift-produced protein isolates were investigated. Even at 2.5 % LPC, the formation of volatile oxidation-derived aldehydes, including hexanal, (E)-2-hexenal, heptanal, octanal, and 2,4-heptadienal, were prevented during the actual protein isolate production. Adding 10 % LPC successfully prevented formation of all these aldehydes also during eight days ice storage which was explained by the partitioning of phenolics, especially ideain (1.09 mg/g dw) and procyanidin A1 (65.5 mg/g dw), into isolates. Although higher amounts of LPC (20-30 %) further prolonged the oxidation lag phase, it reduced total protein yield, increased the consumption of acid and base, and darkened protein isolates. Therefore, it is recommended to use 10 % LPC when pH-shift-processing sensitive fish raw materials as a route to mitigate lipid oxidation and at the same time promote industrial symbiosis and more circular food production.
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The relationship between hemolysis and lipid oxidation was explored in red blood cell (RBCs)-spiked washed cod mince (WCM). At pH 6.8 and 3 ± 1 °C, intact RBCs (71 µM Hb) delayed lipid oxidation by 1 day compared to WCM with partly or fully lysed RBCs which oxidized immediately. Intact RBCs also lowered peak peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) with up to 59.5% and 48.1%, respectively. Adding 3% (v/w) blood plasma to RBC-spiked WCM delayed the lipid oxidation onset from 1 to 3-4 days without delaying hemolysis. At pH 6.4 the oxidation onset in RBC-WCM was the same as for pH 6.8 while at pH 7.2-7.6 lipid oxidation was suppressed for 7 days. Micrographs revealed RBC-lysis from day 2 at pH 6.4 but at pH 7.6, RBC stayed intact for ≥ 7 days. Thus, assuring presence of plasma-derived antioxidants and/or elevating muscle pH to avoid hemolysis can aid valorization of blood rich underutilized fish raw materials.
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Hemólisis , Músculos , Animales , Plasma , Eritrocitos , Peces , Lípidos , Concentración de Iones de HidrógenoRESUMEN
Currently, side-streams (e.g., head, backbone, tail, and intestines) generated in the fish processing industry often end up as low-value products for feed applications or even as waste. In order to upcycle such side-streams, they need to be preserved to avoid oxidative degradation of the lipids between the generation point and the valorization plant. In the cod filleting industry, three main solid side-streams: viscera, heads, and backbones, are obtained. Hence, this study aimed to identify the most efficient antioxidant for preserving the cod side-streams using a dipping-based strategy prior to pre-valorization storage at low temperatures (ice and frozen storage). The dipping solutions evaluated contained: (i) a lipophilic rosemary extract (0.05% and 0.2% in 0.9% NaCl), (ii) Duralox MANC (a mixture of rosemary extract, ascorbic acid, tocopherols, and citric acid; 2% in 0.9% NaCl), and (iii) NaCl (0.9%) w/w solution. One group was not dipped. No dipping and dipping in NaCl were included as controls. The results showed a positive effect of dipping with solutions containing antioxidants as measured by peroxide value (PV), TBA-reactive substances (TBARS), and sensory profiling, e.g., rancid odor. Moreover, the oxidative stability increased with decreased storage temperature. The cod side-streams were in general most efficiently preserved by Duralox MANC, followed by the lipophilic rosemary extract (0.2%), compared to no dipping and dipping in NaCl solution and the lower concentration of the lipophilic rosemary extract (0.05%). The efficiency of the antioxidant treatments was independent of the side-stream fraction and storage temperature. Thus, using antioxidant dipping combined with low temperature storage is an efficient preservation method for maintaining the quality of the lipids in cod solid side-streams during their pre-valorization storage.
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Antioxidantes , Solución Salina , Antioxidantes/farmacología , Temperatura , Cloruro de Sodio , Ríos , Lípidos , Estrés OxidativoRESUMEN
The possibility of reducing the amount of fresh water used during alkaline the pH-shift processing of salmon head (SH) and herring frame (HF) was evaluated with ultrasound (US) as a tool to mitigate its negative effects on protein yield. The role of water ratio and US for homogenate viscosity, mass yield, crude composition, functional properties and lipid oxidation of the SH and HF protein isolates were also investigated. Applying US during the solubilization step of the pH-shift process completely compensated for the reduced protein yield coming from using 3 rather than 6 volumes of water for HF, but not for SH. Using US had no negative effect on the composition and protein functionality of the HF protein isolate. However, it slightly increased its level of secondary lipid oxidation products. Altogether, applying US during the pH-shift processing at low water ratios can be a promising solution for more resource-smart valorization of herring side streams.
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Proteínas de Peces , Salmón , Animales , Ríos , Peces , Proteínas , Concentración de Iones de Hidrógeno , Agua , LípidosRESUMEN
Seaweed is often recognized for its potential health benefits, attributed to its abundance of dietary fibers, protein, and polyphenols. While human observational studies have shown promise, the collective evidence from human intervention trials remains limited. This narrative review aims to comprehensively analyze the effects of seaweed intake on humans, while critically assessing the methodology, including Cochrane risk-of-bias assessment. A search was conducted in online databases, including PubMed, Scopus, and Google Scholar, covering the period from 2000 to May 2023. The focus was on randomized controlled clinical trials (RCTs) evaluating the impact of whole seaweed, either consumed as capsules, integrated into food products or as part of meals. Various health outcomes were examined, including appetite, anthropometric measures, cardiometabolic risk factors, thyroid function, markers of oxidative stress, and blood mineral concentrations. Out of the 25 RCTs reviewed, the findings revealed limited yet encouraging evidence for effects of seaweed on blood glucose metabolism, blood pressure, anthropometric measures, and, to a lesser extent, blood lipids. Notably, these favorable effects were predominantly observed in populations with type-2 diabetes and hypertension. Despite most trials selecting a seaweed dose aligning with estimated consumption levels in Japan, considerable variability was observed in the pretreatment and delivery methods of seaweed across studies. Moreover, most studies exhibited a moderate-to-high risk of bias, posing challenges in drawing definitive conclusions. Overall, this review highlights the necessity for well-designed RCTs with transparent reporting of methods and results. Furthermore, there is a need for RCTs to explore seaweed species cultivated outside of Asia, with a specific emphasis on green and red species. Such studies will provide robust evidence-based support for the growing utilization of seaweed as a dietary component in regions with negligible seaweed consumption, e.g., Europe.
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The overall goal of this study was to provide solutions to innovative microalgae-based technology for wastewater remediation in a cold-water recirculating marine aquaculture system (RAS). This is based on the novel concept of integrated aquaculture systems in which fish nutrient-rich rearing water will be used for microalgae cultivation. The produced biomass can be used as fish feed, while the cleaned water can be reused, to create a highly eco-sustainable circular economy. Here, we tested three microalgae species Nannochloropis granulata (Ng), Phaeodactylum tricornutum (Pt), and Chlorella sp (Csp) for their ability to remove nitrogen and phosphate from the RAS wastewater and simultaneously produce high-value biomass, i.e., containing amino acids (AA), carotenoids, and polyunsaturated fatty acids (PUFAs). A high yield and value of biomass were achieved for all species in a two-phase cultivation strategy: i) a first phase using a medium optimized for best growth (f/2 14x, control); ii) a second "stress" phase using the RAS wastewater to enhance the production of high-value metabolites. Ng and Pt performed best in terms of biomass yield (i.e., 5-6 g of dry weight, DW.L-1) and efficient cleaning of the RAS wastewater from nitrite, nitrate, and phosphate (i.e., 100% removal). Csp produced about 3 g L-1 of DW and reduced efficiently only nitrate, and phosphate (i.e., about 76% and 100% removal, respectively). The biomass of all strains was rich in protein (30-40 % of DW) containing all the essential AA except Methionine. The biomass of all three species was also rich in PUFAs. Finally, all tested species are excellent sources of antioxidant carotenoids, including fucoxanthin (Pt), lutein (Ng and Csp) and ß-carotene (Csp). All tested species in our novel two-phase cultivation strategy thus showed great potential to treat marine RAS wastewater and provide sustainable alternatives to animal and plant proteins with extra added values.
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A novel integrated process for recovery of protein-enriched biomasses from 5% presalting brines and spice brines of herring (Clupea harengus) was investigated by combining carrageenan- and/or acid-driven flocculation (F) plus dissolved air flotation (DAF). The F-DAF technique with carrageenan resulted in protein and lipid recoveries from 5% presalting brine of 78 and 38%, respectively. Without flocculation or with only acidification, protein and lipid recoveries in DAF were only 13 and 10%, respectively. Low protein and lipid recoveries, 8-12 and 1.8-8.2%, respectively, were also obtained when spice brine was subjected to only acidification and DAF. The protein content in dry biomasses from 5% presalting brine and spice brine was 36-43 and 13-16%, respectively. The corresponding lipid levels were 23-31 and 9-18%, respectively, with ash levels of 11-20 and 38-45%, respectively. Biomass proteins contained ≤45% essential amino acids, and the lipids had ≤16% long-chain n-3 polyunsaturated fatty acids. Freeze-dried spice brine biomasses were characterized by anchovy- and spice-related sensory attributes. 5% presalting brine biomasses were connected to fish and seafood attributes and showed gel forming capacity. The outlined F-DAF recovery system can thus recover both nutrients and interesting flavors from the herring process waters, which are currently lost from the food chain.
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The possibility of replacing the very time and resource demanding salting out (SO) method with isoelectric precipitation (IP) during collagen extraction from common starfish and lumpfish was investigated. The effect of IP on yield, structural and functional properties of the collagens was therefore compared with SO. Application of IP resulted in a higher or similar collagen mass yield compared with SO from starfish and lumpfish, respectively. However, the purity of collagens recovered with IP was lower than those recovered with SO. Replacing SO with IP did not affect polypeptide pattern and tropohelical structural integrity of collagen from the two resources as revealed with SDS-PAGE and FTIR analysis. Thermal stability and fibril formation capacity of collagens recovered with IP were also well preserved. Overall, the results showed that the IP can be a promising resource smart alternative for the classic SO precipitation during collagen extraction from marine resources.
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To enable production of high-quality mince from herring backbones, a scalable antioxidant strategy is needed due to the high susceptibility of herring muscle to lipid oxidation. We here measured the stabilizing effect of lab-/pilot-scale predipping of herring backbones (30-500 kg) in antioxidant solutions prior to production of mechanically separated mince (MSM). The antioxidants were (i) Duralox MANC, a mixture of rosemary extract, ascorbic acid, α-tocopherol, and citric acid, and (ii) rosemary extract with or without isoascorbic acid. Delivery of the key rosemary-derived antioxidant components carnosol and carnosic acid was monitored during the dipping process and ice/frozen storage. Predipping in 2% Duralox MANC gave MSM with 26.7-31.7 mg/kg carnosol + carnosic acid and extended the oxidation lag phase from <1 to 12 days during ice storage and from <1 to 6 months during frozen storage compared to control. Dipping in 0.2% rosemary extract with or without 0.5% isoascorbic acid solution gave MSM with 20.6-28.2 mg/kg carnosol + carnosic acid and extended the lag phase to 6 days and 9 months during ice and frozen storage, respectively. Our results confirmed, in pilot scale, that predipping herring coproducts in antioxidant solutions is a promising strategy to utilize these raw materials for, e.g., mince and burger production rather than for low value products as fish meal.
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The seafood industry generates significant amounts of process waters which can generate value upon recovery of their nutrients. Process waters from the herring marination chain and cooking of mussels were here characterized in terms of crude composition, volatile compounds, and nutritional and potentially toxic elements. Protein and total fatty acid contents of herring refrigerated sea water (RSW) reached 3 and 0.14 g/L, respectively, while herring presalting brine (13%) reached 16.3 g/L protein and 0.77 g/L total fatty acid. Among three herring marination brines vinegar brine (VMB), spice brine (SPB), and salt brine (SMB), SPB reached the highest protein (39 g/L) and fatty acids (3.0 g/L), whereas SMB and VMB at the most had 14 and 21 g protein/L, respectively, and 0.6 and 9.9 g fatty acids/L, respectively. Essential amino acid (EAA) in marination brines accounted for up to 59% of total amino acid (TAA). From mussel processing, cooking juice had more protein (14-23 g/L) than the rest of the process waters, and in all water types, EAA reached up to 42% of TAA. For all process waters, the most abundant nutritional elements were Na, K, P, Ca, and Se. The content of all potentially toxic elements was mostly below LOD, except for As which ranged from 0.07 to 1.07 mg/kg among all tested waters. Our findings shed light on liquid seafood side streams as untapped resources of nutrients which can be valorized into food/feed products.
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High acid-consumption and lipid oxidation are challenges when recovering functional proteins from herring co-products via pH-shift-processing. Here, lingonberry press-cake (LP), which is abundant in organic acids and phenolics, was added to alkali-solubilized herring-co-product-proteins (2.5-30 % LP per dry weight) aiming to aid protein precipitation, save hydrochloric acid (HCl) and provide oxidative stability. The results revealed 5-30 % LP addition reduced HCl-consumption by 13-61 % and 19-79 % when precipitating proteins at pH 5.5 and 6.5, respectively. Higher LP% decreased protein content and lightness of protein isolates but raised the lipid content. Precipitation at pH 6.5 used less acid, reduced total protein yield and raised moisture content and darkness of isolates. Contrary to controls, lipid oxidation-derived volatiles did not develop in protein isolates precipitated with 10 % and 30 % LP, neither during the process itself nor during 21 days on ice. Altogether, LP was identified as a promising all-natural processing-aid to use during herring protein isolation.
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The seaweed Saccharina latissima is often blanched to lower iodine levels, however, it is not known how blanching affects protein extraction. We assessed the effect of blanching or soaking (80/45/12 °C, 2 min) on protein yield and protein extract characteristics after pH-shift processing of S. latissima. Average protein yields and extract amino acid levels ranked treatments as follows: blanching-45 °C â¼ control > soaking â¼ blanching-80 °C. Although blanching-45 °C decreased protein solubilization yield at pH 12, it increased isoelectric protein precipitation yield at pH 2 (p < 0.05). The former could be explained by a higher ratio of large peptides/proteins in the blanched biomass as shown by HP-SEC, whereas the latter by blanching-induced lowering of ionic strength, as verified by a dialysis model. Moreover, blanching-45 °C yielded a protein extract with 49 % less iodine compared with the control extract. We recommend blanching-45 °C since it is effective at removing iodine and does not compromise total protein extraction yield.
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Yodo , Phaeophyceae , Aminoácidos , Diálisis Renal , Concentración de Iones de HidrógenoRESUMEN
Effect of catching season (spring vs fall), pre-sorting and selective recombination of different herring filleting co-products on protein yield during valorisation using the alkaline pH-shift technology was studied. Impacts of the pre-processing conditions on lipid oxidation, rheological, structural and functional properties of the proteins were also investigated. The sorted frame fraction resulted in the highest protein yield, myosin content, gel-forming capacity and gel whiteness. pH-shift processing triggered severe proteolytic degradation and lipid oxidation in the head fraction imposing a low-quality protein isolate. The unsorted co-products and the combinations head + frame and head + frame + tail gave protein isolates with gelation and oxidative quality being better than head but below the isolate from frame alone. The spring co-products produced protein isolates with better overall quality than the fall co-products. Altogether, the results revealed the advantage of sorting herring co-products, and the influence of season on protein extraction from herring co-products.
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Proteínas de Peces , Peces , Animales , Proteínas de Peces/química , Concentración de Iones de Hidrógeno , Lípidos , Estaciones del AñoRESUMEN
Cross-processing herring co-products with antioxidant-rich helpers including lingonberry-press-cake, shrimp-shells and seaweed was reported to mitigate lipid oxidation but reduce protein yield. Here, four strategies were used to counteract such yield-reduction; optimizing solubilization/precipitation pH, increasing raw-material-to-water-ratio, replacing single-stage-toothed- by radial-discharge- high-shear-mechanical-homogenization (RD-HSMH) and ultrasonication (US). The effects of RD-HSMH and US on lipid oxidation, protein structural and functional properties were studied. Combining four strategies improved total protein yield by 5-12 %, depending on helper type. More than the confirmed antioxidant effects, cross-processing also improved protein water solubility and emulsification activity but reduced gelation properties. RD-HSMH generally improved protein emulsifying and gelation properties but reduced protein water solubility. US reduced protein water solubility and gelation properties. Altogether, it was recommended for all helpers to increase solubilization pH to 12 and raw-material-to-water-ratio to 1:6 followed by RD-HSMH at 8000 rpm for 90 s, aiming for maximum protein yield and emulsifying and gelation properties.
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Antioxidantes , Alta del Paciente , Animales , Antioxidantes/metabolismo , Peces/metabolismo , Manipulación de Alimentos , Humanos , Concentración de Iones de Hidrógeno , Lípidos , Agua/metabolismoRESUMEN
Weight distribution, proximate composition, fatty acids, amino acids, minerals and vitamins were investigated in five sorted cuts (head, backbone, viscera + belly flap, tail, fillet) emerging during filleting of spring and fall herring (Clupea harengus). The herring co-product cuts constituted â¼ 60 % of the whole herring weight, with backbone and head dominating. Substantial amounts of lipids (5.8-17.6 % wet weight, ww) and proteins (12.8-19.2 % ww) were identified in the co-products, the former being higher in fall than in spring samples. Co-product cuts contained up to 43.1 % long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) of total FA, absolute levels peaking in viscera + belly flap. All cuts contained high levels of essential amino acids (up to 43.3 %), nutritional minerals (e.g., iodine, selenium, calcium, and iron/heme-iron), and vitamins E, D, and B12. Co-products were, in many cases, more nutrient-rich than the fillet and could be excellent sources for both (functional) food and nutraceuticals.
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Meat analogs based on plant protein extracts are rising in popularity as meat consumption declines. A dietary shift away from meat, which has a high iron bioavailability, may have a negative effect on the amount of iron absorbed from the diet. Iron absorption from legumes cultivated in regions not suitable for soy production, such as fava bean, has not yet been explored. The aim of this study was to evaluate non-heme iron absorption from a meal with texturized fava bean protein compared to beef and cod protein meals. The study included two single-blinded iron isotope trials in healthy Swedish women of the ages 18-45 years, each of whom served as their own control. The participants were served matched test meals containing beef and fava bean protein (Study 1) or cod and fava bean protein (Study 2) with radiolabeled non-heme iron 55Fe and 59Fe. The absorption of non-heme iron from test meals was measured by whole-body counting and erythrocyte incorporation. The absorption of non-heme iron, measured as erythrocyte incorporation ratio, from beef protein meal was 4.2 times higher compared to texturized fava bean meal, and absorption from cod protein meal was 2.7 times higher compared to the fava bean meal. The adjusted non-heme iron absorption, normalized to a 40% reference dose uptake, was 9.2% for cod protein meal, 21.7% for beef protein meal, and 4.2% for texturized fava bean meal. A fava bean protein meal has markedly lower iron bioavailability in healthy females compared with a meal of beef or cod protein. Therefore, a dietary shift from meat and fish protein to fava bean protein may increase the risk of iron deficiency.
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Fabaceae , Vicia faba , Adolescente , Adulto , Animales , Disponibilidad Biológica , Bovinos , Femenino , Humanos , Absorción Intestinal , Hierro/metabolismo , Comidas , Persona de Mediana Edad , Ensayos Clínicos Controlados Aleatorios como Asunto , Vicia faba/metabolismo , Adulto JovenRESUMEN
Lipid oxidation is a complex process in muscle-based foods (red meat, poultry and fish) causing severe quality deterioration, e.g., off-odors, discoloration, texture defects and nutritional loss. The complexity of muscle tissue -both composition and structure- poses as a formidable challenge in directly clarifying the mechanisms of lipid oxidation in muscle-based foods. Therefore, different in vitro model systems simulating different aspects of muscle have been used to study the pathways of lipid oxidation. In this review, we discuss the principle, preparation, implementation as well as advantages and disadvantages of seven commonly-studied model systems that mimic either compositional or structural aspects of actual meat: emulsions, fatty acid micelles, liposomes, microsomes, erythrocytes, washed muscle mince, and muscle homogenates. Furthermore, we evaluate the prospects of stem cells, tissue cultures and three-dimensional printing for future model system development. Based on this reviewing of oxidation models, tailoring correct model to different study aims could be facilitated, and readers are becoming acquainted with advantages and shortcomings. In addition, insight into recent technology developments, e.g., stem cell- and tissue-cultures as well as three-dimensional printing could provide new opportunities to overcome the current bottlenecks of lipid oxidation studies in muscle.
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Hemoglobin (Hb) is a powerful promoter of lipid oxidation, particularly in muscle of small pelagic fish species and fish by-products, both having high Hb-levels and highly unsaturated lipids. As Hb is located within the red blood cells (RBCs) it is here hypothesized that the perishable polyunsaturated fatty acids (PUFAs) can be protected from oxidation by limiting hemolysis during early fish processing. Using a model system consisting of washed-resuspended trout (Oncorhynchus mykiss) RBCs (wr-RBCs), the aim of this study was to evaluate how RBC lysis under cold storage was affected by selected parameters linked to blood or muscle: bacterial growth, energy status, pH, RBC membrane lipid oxidation and colloidal osmotic pressure (COP). The results indicated that bacterial growth had a modest effect on hemolysis while pH-values typical for post mortem fish muscle (6.4-6.8), and absence of glucose or albumin stimulated hemolysis. The rapid hemolysis observed at pH 6.4-6.8 correlated with lipid oxidation of the RBC membrane, while the lower hemolysis at pH 7.2-8.0 occurred with low, or without any RBC membrane lipid oxidation. When hemin was added to the RBCs at pH 6.8 hemolysis was induced without parallel RBC membrane oxidation, pointing at Hb-autoxidation and hemin-release per se as important events triggering lysis in fish muscle. Altogether, the study provided valuable findings which ultimately can aid development of new tools to combat lipid oxidation in post mortem fish muscle by limiting hemolysis.
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Hemólisis , Oncorhynchus mykiss , Animales , Eritrocitos , Ácidos Grasos Insaturados/farmacología , Hemina/farmacología , Hemoglobinas/farmacología , Lípidos de la Membrana/farmacología , MúsculosRESUMEN
In this study, ensilaging of herring (Clupea harengus) filleting co-products was taken from lab-scale to pilot scale (1500 L) while monitoring the protein degree of hydrolysis (DH) and lipid oxidation. Subsequently, the possibility of recovering fish oil and protein hydrolysates using batch centrifugation at different g-forces/times was investigated. Around 38% DH was recorded after 2-day pilot-scale ensilaging of herring co-products at ambient temperature (i.e., ~ 22 °C), which was similar to the DH found in lab-scale (40% after 2 days; 22 °C). The lipid oxidation marker 2-thiobarbituric acid reactive substances (TBARS) reached 20 µmole TBARS/kg silage after 2-day ensilaging. Centrifugation of the silage at 3000-8500 × g for 2-20 min revealed successful separation into fish oil and protein hydrolysates. Heat-treating the silage (85 °C; 30 min) prior to centrifugation resulted in significantly higher oil and hydrolysates recoveries; the same being true for increased g-force. At 8500 × g, the recovery of oil and hydrolysates were 9.7 and 53.0% w/w, respectively, from heat-treated silage, while recoveries were 4.1 and 48.1% w/w, respectively, from non-heat treated silage. At 4500 × g, being a more scalable approach, corresponding numbers were 8.2 and 47.1% (w/w) as well as 2.0 and 40.2% (w/w). The recovered fish oil contained 8% EPA and 11% DHA of total fatty acids. Free fatty acids (FFA), peroxide value (PV), p-anisidine value (p-AV), and total oxidation (TOTOX) values of oils were in the range of 4-7% (FFA), 3.6-3.7 meq/kg oil (PV), 2.5-4.0 (p-AV), and 9.9-11.1 (TOTOX), respectively, which were within the acceptable limits for human consumption specified by the GOED voluntary monograph. The recovered protein hydrolysates contained peptides in the molecular weight range 0.3-6 kDa (~ 37%) and 11-34 kDa (~ 63%). Also, the remaining solids contained 15-17% (w/w) protein, having 44-45% essential amino acids. Overall, the results suggest that herring co-product silage is a valuable source of fish oil and protein hydrolysates, paving the way for ensilaging based-biorefining of herring co-products into multiple products. Supplementary Information: The online version contains supplementary material available at 10.1007/s11947-022-02870-9.
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Processes currently used for collagen extraction are complicated requiring a great deal of time and chemicals. Here, high shear mechanical homogenization (HSMH) and ultrasound (US) were integrated in the pretreatment step of collagen extraction from common starfish to reduce chemical use and time consumption. Effects of the assistant technologies on yield, structural integrity and functionality of collagen were also investigated. HSMH reduced the deproteinization time from 6 h to 5 min and its required amount of alkali 4 times, compared with classic methods. HSMH + US reduced the demineralization time from 24 h to 12 h and improved its efficiency in extraction of minerals. Collagen extraction with HSMH and HSMH + US resulted in similar yield as the classic method and did not affect triple helical structural integrity, polypeptide pattern, thermal stability or fibril-formation capacity of the collagens. Altogether, HSMH and US can effectively improve resource efficiency during collagen extraction without imposing negative effect on collagen quality.