Combined effects of isolation temperature and pH on functionality and beany flavor of pea protein isolates for meat analogue applications.

The combined effects of isolation temperature (20, 30 and 40 °C) and pH (2.0-12.0) on yield, techno-functional properties, and beany flavor of pea protein isolates were investigated. Increasing pH from 2.0 to 9.5 and 11.0 increased yields from 37 % to 75 % and 79 %, respectively, at 20 °C. At a constant pH, increasing temperature from 20 to 40 °C did not increase protein recovery; rather, negatively affected the techno-functional properties such as protein solubility, foaming and gelation. Protein isolated at pH 11.0 (20 °C) provided a higher fat absorption, gelation capacity, gel hardness, cohesiveness, chewiness, and gumminess than at pH 9.5, due to higher protein denaturation as supported by their higher surface hydrophobicity. Volatile beany flavor marker hexanal was predominant in all isolates than the starting material, irrespective of isolation temperature, probably due to lipid oxidation. The results provide a basis for tuning the isolation process for producing pea protein isolates with desired techno-functional properties for meat analogue applications.

Pilot-Scale Ensilaging of Herring Filleting Co-Products and Subsequent Separation of Fish Oil and Protein Hydrolysates.

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.

Effect of antioxidants on lipid oxidation in herring (Clupea harengus) co-product silage during its production, heat-treatment and storage.

Provided high product quality, ensilaging can be used to valorize fish filleting co-products into a silage suitable for food applications. However, a documented challenge for products from hemoglobin-rich fish raw materials is the high susceptibility to lipid oxidation, calling for stabilization by antioxidants. In a comparison among different rosemary-containing antioxidants and isoascorbic acid, we here found that the commercial mixture Duralox MANC-213 (MANC) provided the best protection against peroxide value and 2-thiobarbituric acid reactive substances (TBARS) development during ensilaging of herring filleting co-products (0-7 days, 22 °C), and also during subsequent heat-treatment (30 min, 85 °C). Increasing MANC concentration from 0.25 and 0.75 to 1.25% lowered TBARS values from 43.53 and 25.12 to 18.04 µmole TBARS/Kg silage, respectively, after 7 days of ensilaging. During storage at 4 °C/22 °C in presence of MANC, 1.25% provided the highest protection with 87-90% and 66-73% lower TBARS, at 4 °C and 22 °C, respectively, at 6 months compared to the controls. At this time point, heat-treated silages had lower protein degree of hydrolysis and free amino acids values than the non-heat-treated one. Regardless of antioxidant addition, total volatile basic nitrogen (TVB-N) formation still increased during the storage, but, overall, TVB-N values in silages were below the acceptable limit of 30 mg TVB-N/100 g fish for human consumption. Together with lipid oxidation data, this suggest that herring silage produced in presence of antioxidants can be used both for high quality feed and food applications.

Hemoglobin-mediated lipid oxidation of herring filleting co-products during ensilaging and its inhibition by pre-incubation in antioxidant solutions.

The aims of this study were to investigate the role of hemoglobin (Hb) in lipid oxidation development during ensilaging of herring filleting co-products, and, to inhibit this reaction by pre-incubating the co-products in water or physiological salt, with/without different antioxidants. Results showed that both peroxide value (PV) and 2-thiobarbituric acid reactive substances (TBARS) gradually increased during 7 days of ensilaging at 22 °C in absence of antioxidants. The increase in TBARS was proportional to the Hb levels present, while PV was less affected. A Hb-fortified Tris-buffer model system adjusted to pH 3.50 confirmed that Hb changed immediately from its native oxyHb to the metHb state, which facilitated heme group release and thus probably explains the increased PV and TBARS during ensilaging. Pre-incubating the co-products for 30 s in a solution containing 0.5% rosemary extract was the most promising strategy to inhibit lipid oxidation both in the co-products during pre-processing storage and during the actual ensilaging. The solution could be re-used up to ten times without losing its activity, illustrating that this methodology can be a scalable and cost-effective strategy to extend the oxidative stability of herring co-products allowing for further value adding e.g., into a high-quality silage.

Understanding the effect of temperature and time on protein degree of hydrolysis and lipid oxidation during ensilaging of herring (Clupea harengus) filleting co-products.

The aims of this study were to investigate the effect of temperature, time and stirring on changes in protein degree of hydrolysis (DH), free amino acids (FAA), lipid oxidation and total volatile basic nitrogen (TVB-N) during ensilaging of herring (Clupea harengus) filleting co-products. Results showed that temperature and time, and in some cases the interaction effect between these two factors, significantly influenced all the studied responses. Increasing ensilaging temperature and time from 17 to 37 °C and 3 to 7 days, respectively, increased DH, FAA, and TVB-N content from 44.41 to 77.28%, 25.31 to 51.04 mg/g, and 4.73 to 26.25 mg/100 g, respectively. The lipid oxidation marker 2-thiobarbituric acid reactive substances (TBARS) did not increase with time at temperatures above 22 °C, while 2-pentylfuran increased up to 37 °C. Based on the process parameters and responses investigated in this study, and considering energy requirements, it was suggested to perform ensilaging at ambient temperatures (i.e. around 20 °C) with continuous stirring at 10 rpm for 1-3 days; the exact length being determined by the desired DH.

Valorization of Brewer's spent grain to prebiotic oligosaccharide: Production, xylanase catalyzed hydrolysis, in-vitro evaluation with probiotic strains and in a batch human fecal fermentation model.

Brewer's spent grain (BSG) accounts for around 85% of the solid by-products from beer production. BSG was first extracted to obtain water-soluble arabinoxylan (AX). Using subsequent alkali extraction (0.5 M KOH) it was possible to dissolve additional AX. In total, about 57% of the AX in BSG was extracted with the purity of 45-55%. After comparison of nine xylanases, Pentopan mono BG, a GH11 enzyme, was selected for hydrolysis of the extracts to oligosaccharides with minimal formation of monosaccharides. Growth of Bifidobacterium adolescentis (ATCC 15703) was promoted by the enzymatic hydrolysis to arabinoxylooligosaccharides, while Lactobacillus brevis (DSMZ 1264) utilized only unsubstituted xylooligosaccharides. Furthermore, utilization of the hydrolysates by human gut microbiota was also assessed in a batch human fecal fermentation model. Results revealed that the rates of fermentation of the BSG hydrolysates by human gut microbiota were similar to that of commercial prebiotic fructooligosaccharides, while inulin was fermented at a slower rate. In summary, a sustainable process to valorize BSG to functional food ingredients has been proposed.