Effect of κ-carrageenan on gelation and gel characteristics of Antarctic krill (Euphausia superba) protein isolated with isoelectric solubilization/precipitation.

Isoelectric solubilization/precipitation (ISP) was used to extract krill protein isolate (KPI). Due to krill endogenous proteases ISP-KPI barely forms gel which is a major hurdle in wide application of this tremendous protein source for direct human consumption. The objective was to improve gelation and elucidate interaction mechanism between κ-carrageenan and KPI. κ-Carrageenan was added to KPI at 0.00, 0.75, 1.50, 2.25, and 3.00 g/150 g and heated at 90 °C for 15 min. Added κ-carrageenan improved texture of KPI gels. However, the level of addition had minimal effect. SDS-PAGE revealed severe proteolysis of KPI even during heating. Total and free sulfhydryl were not different, while surface hydrophobicity and water-holding-capacity slightly decreased and increased, respectively with added κ-carrageenan. Fourier-transform-infrared-spectroscopy showed none-to-minimal shift of ß-sheet and α-helical protein structure; while CC and CO stretching as well as CH bending of κ-carrageenan showed the greatest shift, indicating that κ-carrageenan was mainly responsible for gel formation.

Micro-emulsification/encapsulation of krill oil by complex coacervation with krill protein isolated using isoelectric solubilization/precipitation.

This study determined feasibility of krill protein isolated with isoelectric solubilization/precipitation (ISP) as wall material to microencapsulate krill oil by freeze-drying. Effects of krill oil/krill protein ratio on properties of microcapsules were investigated. With increased ratio, crude protein of microcapsules decreased, while total lipid increased. Although microcapsule oil loading capacity increased, loading and encapsulation efficiencies decreased. Thin layer chromatography (TLC) confirmed abundance of phospholipids, which are amphiphilic; and thus, resulted in stable emulsion (emulsion stability index). Microcapsules contained ω-3 polyunsaturated fatty acids (PUFAs) at 43-60, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) at 28-41 and 9-11 g/100g of total FAs, respectively. SDS-PAGE electrophoresis revealed proteolysis of ISP krill protein, probably causing reduced loading and encapsulation efficiencies. SEM showed that krill oil/krill protein ratio affected surface microstructure. ISP krill protein showed potential as a wall material to microencapsulate krill oil; and thus, expand application of krill oil/protein for human consumption.

Storage stability of egg sticks fortified with omega-3 fatty acids.

BACKGROUND: Egg sticks fortified with omega-3 polyunsaturated fatty acids (ω-3 PUFAs) were developed by replacing egg yolk with salmon, algae, and flax oils. Egg sticks were cooked before analysis. Quality indicators for storage stability under different packaging and temperature were determined throughout a 28-day storage. Egg sticks were vacuum and non-vacuum packed. Further, both packaging treatments were divided into two storage temperatures of 4 and 10 °C. Quality indicators were determined every 7 days, including pH, syneresis, texture, color, microbial growth, proximate composition, fatty acid profile, and lipid oxidation. RESULTS: Vacuum-packed egg sticks stored at 4 °C had slower degradation over time than all other treatments; however, they also had higher syneresis, harder texture, and higher anaerobic growth. Although vacuum packaging slowed lipid oxidation, it had limited effect on prevention of ω-3 PUFAs degradation; whereas refrigeration (4 °C) seemed to prevent degradation of ω-3 PUFAs better than it could slow lipid oxidation. CONCLUSION: Based on the results, it can be concluded that both vacuum packaging and refrigeration at 4 °C decrease degradation of egg sticks developed in the present study during storage. Under these conditions, egg sticks may maintain stability for at least 21 days of storage. © 2017 Society of Chemical Industry.

pH shift protein recovery with organic acids on texture and color of cooked gels.

BACKGROUND: Isoelectric solubilization and precipitation (ISP) processing uses pH shifts to separate protein from fish frames, which may increase commercial interest for silver carp. Texture and color properties of gels made from silver carp protein recovered at different pH strategies and organic acid types were compared. ISP was applied to headed gutted silver carp using 10 mol L(-1) sodium hydroxide (NaOH) and either glacial acetic acid (AA) or a (1:1) formic and lactic acid combination (F&L). Protein gels were made with recovered protein and standard functional additives. RESULTS: Texture profile analysis and the Kramer shear test showed that protein gels made from protein solubilized at basic pH values were firmer, harder, more cohesive, gummier and chewier (P < 0.05) than proteins solubilized under acidic conditions. Acidic solubilization led to whiter (P < 0.05) gels, and using F&L during ISP yielded whiter gels under all treatments (P < 0.05). CONCLUSION: Gels made from ISP-recovered silver carp protein using organic acids show potential for use as a functional ingredient in restructured foods.

Physicochemical properties of surimi gels fortified with dietary fiber.

Although dietary fiber provides health benefits, most Western populations have insufficient intake. Surimi seafood is not currently fortified with dietary fiber, nor have the effects of fiber fortification on physicochemical properties of surimi been thoroughly studied. In the present study, Alaska pollock surimi was fortified with 0-8 g/100 g of long-chain powdered cellulose as a source of dietary fiber. The protein/water concentrations in surimi were kept constant by adding an inert filler, silicon dioxide in inverse concentrations to the fiber fortification. Fiber-fortified surimi gels were set at 90 °C. The objectives were to determine (1) textural and colour properties; (2) heat-induced gelation (dynamic rheology); and (3) protein endothermic transitions (differential scanning calorimetry) of surimi formulated with constant protein/water, but variable fiber content. Fiber fortification up to 6 g/100 g improved (P<0.05) texture and colour although some decline occurred with 8 g/100g of fiber. Dynamic rheology correlated with texture and showed large increase in gel elasticity, indicating enhanced thermal gelation of surimi. Differential scanning calorimetry showed that fiber fortification did not interfere with thermal transitions of surimi myosin and actin. Long-chain fiber probably traps water physically, which is stabilized by chemical bonding with protein within surimi gel matrix. Based on the present study, it is suggested that the fiber-protein interaction is mediated by water and is physicochemical in nature.

Effect of electron beam on chemical changes of nutrients in infant formula.

Infant milk formula has recently been implicated as a transmission vehicle for an emerging foodborne pathogen, Enterobacter sakazakii, resulting in high mortality rates. Electron beam (e-beam) efficiently and non-thermally inactivates foodborne pathogens, including E. sakazakii, in infant milk formula. However, the effects of e-beam on chemical changes of nutrients in infant formula have not been determined. Therefore, the objective of this study was to fulfill this gap. Dehydrated infant milk formula was processed with e-beam at 0 (control) to 25 kGy. Amino acid, fatty acid, and mineral profiles (AAP, FAP, and MP, respectively), as well as protein degradation and lipid oxidation, were determined. There were no differences (P>0.05) in FAP, AAP, and MP. SDS-PAGE electrophoresis qualitatively detected three major protein bands in all samples up to 25 kGy. Densitometry analysis of SDS-PAGE gels confirmed no size degradation (P>0.05) as a function of increased e-beam dose. Totol-volatile-basic-nitrogen (TVBN) excluded (P>0.05) protein degradation due to microbial activity. There was no increase (P>0.05) in lipid oxidation, as assessed with thiobarbituric-reactive-substances (TBARS), except in samples processed at 25 kGy. Dehydrated formula has low water activity, which likely protected nutrients from e-beam-induced chemical changes. This study demonstrates that proteins, lipids, and minerals in infant milk formula are stable when processed with e-beam up to 25 kGy at low temperature and under anaerobic conditions.

Interactions of dietary fibre and omega-3-rich oil with protein in surimi gels developed with salt substitute.

Most Western populations have insufficient intake of fibre and ω-3 polyunsaturated fatty acids (PUFAs), while sodium intake greatly exceeds the recommended maximum. Surimi seafood is not currently fortified with these nutraceutical ingredients. Alaska pollock surimi seafood was developed with salt substitute and fortified with either 6g/100g of fibre or 10 g/100g of ω-3 oil (flax:algae:menhaden, 8:1:1) or fibre+ω-3 oil (6g/100g of fibre+10 g/100g of ω-3 oil). The objective was to determine effects of the dietary fortification on physicochemical properties of surimi. Fortification with either dietary fibre or ω-3 oil alone or in combination enhanced (P<0.05) rheological and textural characteristics. The combined fortification had a synergistic effect on rheological properties. This indicates greater gelation of surimi in the presence of fibre+ω-3 oil, suggesting their interaction with surimi myofibrillar proteins. Fibre results in protein dehydration increasing protein concentration; while oil is immobilised by protein filling void spaces in the gel matrix. Differential scanning calorimetry showed that fibre and ω-3 oil did not interfere with normal denaturation of surimi proteins. Colour properties were only slightly affected (P<0.05). Fortification of surimi with fibre and ω-3 oil resulted in a quality product that could be useful in developing surimi products with nutritional benefits.

Chemical properties of ω-3 fortified gels made of protein isolate recovered with isoelectric solubilisation/precipitation from whole fish.

Protein isolate was recovered from whole gutted fish using isoelectric solubilisation/precipitation (ISP). The objective was to determine chemical properties of heat-set gels made of the ISP protein isolate fortified with ω-3 polyunsaturated fatty acids (PUFAs)-rich oils (flaxseed, fish, algae, krill, and blend). The extent of the PUFAs increase, ω-6/ω-3 FAs and unsaturated/saturated FAs ratios, and the indices of thrombogenicity and atherogenicity depended on specific ω-3 PUFAs-rich oil used to fortify protein isolate gels. Lipid oxidation in ω-3 PUFAs fortified gels was minimal, although greater (P<0.05) than control gels (without ω-3 PUFAs fortification). However, all gels were in the slightly rancid, but acceptable range. The commonly used thiobarbituric-acid-reactive-substances (TBARS) assay to determine lipid oxidation in seafood may be inaccurate for samples containing krill oil due to its red pigment, astaxanthin. Protein degradation (total-volatile-basic-nitrogen) was greater (P<0.05) in ω-3 PUFAs fortified gels than control gels. However, all gels were considerably below the acceptability threshold for protein degradation. The shear stress of ω-3 PUFAs fortified gels was generally greater than the control gels and the shear strain was generally unchanged. This study demonstrates that ω-3 PUFAs fortification of protein isolates recovered with ISP from fish processing by-products or whole fish has potential application in the development of functional foods.

Compositional characteristics of materials recovered from headed gutted silver carp (Hypophthalmichthys molitrix) by isoelectric solubilization and precipitation using organic acids.

UNLABELLED: Protein was recovered from headed gutted silver carp by isoelectric solubilization at pH 2.5, 3.0, 11.5, or 12.0 and precipitation (ISP) at pH 5.5 using acetic (AA) or a 30% formic and lactic acid combination (F&L) and 10 N sodium hydroxide. Total protein and fat recovery yields, proximate composition and mineral analyses of fractions were determined. Protein and lipid recovery yields when solubilized under basic conditions were comparable to yields reported from other studies using hydrochloric acid; however, the recovered fractions were less pure. Processing at basic pH using AA was more effective than F&L at removing impurities (P < 0.05) from the recovered protein fraction and impurities were effectively removed from recovered lipids regardless of processing pH or acid type (P > 0.05). For the most part, sodium was greater (P < 0.05) and there was less calcium, phosphorus, magnesium, and iron (P < 0.05) in the recovered protein regardless of acid used when compared to the initial paste. This research shows that organic acids have the potential to recover protein and lipid by ISP processing. PRACTICAL APPLICATION: This research presents a reliable method for extracting nutritionally valuable fish protein and oils from otherwise hard to process fish and its byproducts. Replacing the traditionally used strong acids with organic acids might further accomplish bacterial load reduction while resulting in similar to or improved protein recovery yields. Therefore, this technology may increase the commercial viability of hard to process fish.

Isoelectric solubilization/precipitation as a means to recover protein isolate from striped bass (Morone saxatilis) and its physicochemical properties in a nutraceutical seafood product.

Excessive dietary intake of Na (i.e., NaCl) contributes to hypertension, which is a major risk factor for cardiovascular disease. Normally, NaOH and HCl are used to dissolve and precipitate, respectively, fish muscle proteins in isoelectric solubilization/precipitation (ISP), therefore contributing to increased Na content in the recovered fish protein isolates (FPI). Substitution of NaOH with KOH may decrease the Na content in FPI and, thus, allow development of reduced-Na seafood products. In this study, FPI was recovered with ISP using NaOH or KOH. In order to develop a nutraceutical seafood product, the FPI was extracted with NaCl or KCl-based salt substitute and subjected to cold- or heat-gelation. In addition, standard nutraceutical additives (ω-3 fatty acids-rich oil and dietary fiber) along with titanium dioxide (TiO2) were added to FPI. Color, texture, dynamic rheology, Na and K content, and lipid oxidation of the FPI gels were compared to commercial Alaska pollock surimi gels. FPI gels had greater (p < 0.05) whiteness, good color properties (L*a*b*), and generally better textural properties when compared to surimi gels. Although the ISP-recovered FPI and surimi developed similar final gel elasticity, the proteins in FPI and surimi had different gelation pattern. A reduction (p < 0.05) of Na content and simultaneous increase (p < 0.05) in K content of FPI gels was achieved by the substitution of NaOH with KOH during ISP and NaCl with the KCl-based salt substitute during formulation of the FPI paste. Although cooking and addition of NaCl during formulation of the FPI paste increased (p < 0.05) lipid oxidation in FPI gels, TBARS values were much below rancidity levels. These results indicate that KOH can replace NaOH to recover FPI from whole gutted fish for subsequent development of nutraceutical seafood products tailored for reduction of diet-driven cardiovascular disease.

A three-prong strategy to develop functional food using protein isolates recovered from chicken processing by-products with isoelectric solubilization/precipitation.

BACKGROUND: Skin-on bone-in chicken drumsticks were processed with isoelectric solubilization/precipitation to recover muscle proteins. The drumsticks were used as a model for dark chicken meat processing by-products. The main objective of this study was conversion of dark chicken meat processing by-products to restructured functional food product. An attempt was made to develop functional food product that would resemble respective product made from boneless skinless chicken breast meat. A three-prong strategy to address diet-driven cardiovascular disease (CVD)with a functional food was used in this study. The strategy included addition of three ingredients with well-documented cardiovascular benefits: (i) ω-3 polyunsaturated fatty acid-rich oil (flaxseed-algae, 9:1); (ii) soluble fiber; and (iii) salt substitute. Titanium dioxide, potato starch, polyphosphate, and transglutaminase were also added. The batters were formulated and cooked resulting in heat-set gels. RESULTS: Color (L*a*b*), texture (torsion test, Kramer shear test, and texture profile analysis), thermal denaturation (differential scanning calorimetry), and gelation (dynamic rheology) of chicken drumstick gels and chicken breast gels were determined and compared. Chicken drumstick gels generally had comparable color and texture properties to the gels made from chicken breast meat. The endothermic transition (thermal denaturation) of myosin was more pronounced and gelation properties were better for the drumstick gels. CONCLUSION: This study demonstrated a feasibility to develop functional food made of muscle proteins recovered with isoelectric solubilization/precipitation from low-value dark chicken meat processing by-products. The functional food developed in this study was enriched with CVD-beneficial nutrients and had comparable instrumental quality attributes to respective products made of chicken breast meat. Although the results of this study point towards the potential for a novel, marketable functional food product, sensory tests and storage stability study are recommended.

Characterization of lipids and antioxidant capacity of novel nutraceutical egg products developed with omega-3-rich oils.

BACKGROUND: Cardiovascular disease has had an unquestioned status of the number one cause of death in the US since 1921. Omega-3 polyunsaturated fatty acids (ω-3 PUFAs) have cardio-protective benefits. However, egg is typically a poor source of ω-3 PUFAs and, in general, the American diet is low in these cardio-protective fatty acids. Novel, nutritionally enhanced egg products were developed by substituting yolk with ω-3 PUFA-rich flaxseed, menhaden, algae, or krill oil. Experimental egg products matched composition of hen egg (whole egg). The experimental egg products, mixed whole egg, and a liquid egg product (Egg Beaters) were microwave-cooked and compared. RESULTS: Although fat, protein, and moisture contents of experimental egg products matched (P > 0.05) mixed whole egg, experimental egg products had more (P < 0.05) ω-3 PUFAs, lower (P < 0.05) ω-6/ω-3 ratio, and depending on oil added, a higher (P < 0.05) unsaturated/saturated fatty acids ratio compared to mixed whole egg. Triglycerides were the main lipid class in all experimental egg products except those developed with krill oil, which had even more phospholipids than mixed whole egg. Analysis of thiobarbituric acid reactive substances showed that lipid oxidation of experimental egg products was lower (P < 0.05) or similar (P > 0.05) to mixed whole egg, except for experimental egg products with krill oil. However, peroxide value showed that all egg samples had minimal oxidation. Experimental egg products developed with menhaden or flaxseed oil had the highest (P < 0.05) concentration of the antioxidant, ethyoxquin compared to all other egg samples. However, experimental egg products with krill oil likely contained a natural antioxidant, astaxanthin. CONCLUSION: This study demonstrated an alternative approach to developing novel, nutraceutical egg products. Instead of dietary modification of chicken feed, yolk substitution with ω-3 PUFAs oils resulted in enhancement of ω-3 PUFAs beyond levels possible to achieve by modifying chicken feed.

Radioresistance development of DNA repair deficient Escherichia coli DH5α in ground beef subjected to electron beam at sub-lethal doses.

PURPOSE: Electron beam (e-beam) efficiently and non-thermally inactivates microorganisms in food by lethal DNA changes (direct effects) and free radicals from water radiolysis (in-direct effects). Non-pathogenic Escherichia coli DH5α (α substrain of DH5 described by Hanahan 1985 , 'DH' stands for Douglas Hanahan) is a microorganism that lacks DNA repair capability, resulting in high radiosensitivity. Studying microbial inactivation of E. coli DH5α repeatedly subjected to sub-lethal e-beam in ground beef may enhance understanding of microbial radioresistance. The objective of this study was to determine if repetitive processing with e-beam at sub-lethal doses increases D-value (e-beam dose required to inactivate one log of microbial population) of E. coli DH5α in ground beef. MATERIALS AND METHODS: Survivors from the highest e-beam dose were isolated and incubated in ground beef for the next cycle of e-beam processing. Five cycles were conducted. To acclimatise E. coli DH5α, first two cycles used low doses. D-values were determined following the third cycle. RESULTS: D-values increased (p < 0.05) significantly with each cycle. Thus, E. coli DH5α has a capability to develop greater radioresistance under these experimental conditions. Following the third cycle D-values were 0.32 ±â€Š0.006 and 0.32 ±â€Š0.002 kGy for survivors enumerated on non-selective and selective media, respectively; the fourth cycle 0.39 ±â€Š0.007 and 0.40 ±â€Š0.019 kGy; and the fifth cycle 0.46 ±â€Š0.006 and 0.46 ±â€Š0.020 kGy. D-values on non-selective and selective media were similar (p > 0.05) indicating absence of cell recovery in E. coli DH5α. CONCLUSIONS: E. coli DH5α increases radioresistance to e-beam as a result of repetitive exposure to sub-lethal doses despite its DNA repair deficiency.

The effect of a flaxseed oil-enhanced diet on the product quality of farmed brook trout (Salvelinus fontinalis) fillets.

The effects of dietary modification with flaxseed oil-enhanced (Flax) feed on the product quality of brook trout fillets were examined. Trout were fed a commercial feed supplemented with fish oil (CD) or flaxseed oil (Flax) for 165 d before harvesting. Proximate composition and fatty acid profile were determined on fillets. Quality parameters of the raw fillets were examined over the storage period by measuring color (L*, a*, b*), muscle pH, and thiobarbituric acid reactive substances test. Evaluations on the cooked fillets included sensory evaluation with triangle tests and a paired preference test. There were no differences in proximate composition between the groups; however, the total omega-3 fatty acids were greater in Flax fillets (P<0.05). Diet and day were shown to interact in their effect on whiteness, pH, and lipid oxidation (P<0.05); however, linear regression did not determine that malondialdehyde concentration was associated with time in either diet type implying that lipid oxidation in the vacuum-packed fish was controlled at storage temperatures (4°C). Sensory panelists were able to choose the odd sample in a replicated triangle test analyzed using the ß-binomial model, and there was preference for Flax fillets (P<0.05). Results indicate that a Flax-enhanced diet would have favorable effects on product quality of farmed brook trout.

Recovery of Salmonella enterica serovars Typhimurium and Tennessee in peanut butter after electron beam exposure.

UNLABELLED: The effect of electron beam (e-beam) radiation on the recovery of Salmonella serotypes Tennessee (ATCC 10722) and Typhimurium (ATCC 14028) in creamy peanut butter over a 14-d storage period at 22 °C was studied. Each Salmonella type was independently inoculated into peanut butter and subjected to e-beam doses that ranged from 0 to 3.1 kGy, confirmed by film dosimetry. After 2-, 4-, 6-, 8-, and 14-d of storage, microbial analyses were conducted. Survivors were recovered on growth and selective media using standard spread-plating methods. Microbial counts (CFU/g) were log-converted and differences were determined by ANOVA and Tukey's Honestly Significant Differences test. When samples were not e-beam-treated, there were no significant changes (P > 0.05) in microbial numbers over time. In e-beamed samples, microbial numbers decreased over time; however, reductions were not always significant. Initial recovery rates (R-rates) 2 d after e-beam treatment were significantly different for the 2 strains of Salmonella and between recovery media (P < 0.05); however, these differences did not persist for the remainder of the storage period (P > 0.05) indicating that injured cells were not able to survive in the high-fat, low-water activity peanut butter environment. R-rates for both strains of Salmonella were maintained until day 14 when there were significant reductions in Salmonella Typhimurium (P < 0.05). These results indicate that Salmonella Tennessee and Salmonella Typhimurium will survive in peanut butter when exposed to nonlethal doses of e-beam irradiation. PRACTICAL APPLICATION: Electron beam (e-beam) irradiation is an alternative to thermal processing; this technique inactivates microorganisms and insects that might be present in a food by generating radiation by accelerated electrons that inactivate organisms directly because of interaction with cell components and indirectly by producing free radicals that disrupt integrity of the cell membrane. E-beam radiation will reduce the number of probable microbiological hazards that could be present while the food remains generally unaffected in texture, taste, and nutritional value. A recent study showed e-beam irradiation to be effective at reducing both Salmonella Tennessee and Typhimurium in peanut butter by one log after exposure to less than 1 kGy, highlighting the need to explore this process further.

Compositional characteristics of materials recovered from whole gutted silver carp (Hypophthalmichthys molitrix) using isoelectric solubilization/precipitation.

Isoelectric solubilization/precipitation (ISP) at acidic and basic pH was applied to whole carp, yielding proteins, lipids, and insolubles. The objective was to characterize composition of recovered materials. Crude protein was concentrated to 89-90% in proteins recovered at acidic pH and to 94-95% at basic pH. Basic pH yielded proteins with more (P < 0.05) essential amino acids (EAAs). EAA content in recovered proteins met FAO/WHO/UNO requirements. ISP did not affect fatty acid (FA) composition. Lipids recovered at acidic pH contained 88-89% of total fat and at basic pH, 94-97%. Total fat in recovered proteins was low, with EPA and DHA at the highest (P < 0.05) percentage for pH 11.5. ISP, particularly basic pH, effectively removed impurities such as bones and scales from whole carp. This is indicated by 3.8-5.8% of ash in recovered proteins compared to 11.2% for whole carp and 5.4% for boneless/skinless carp fillets. Basic pH yielded less (P < 0.05) Ca, P, and Mg in recovered proteins. These minerals were more (P < 0.05) concentrated in insolubles recovered with basic pH. This study indicates that materials recovered from whole carp using ISP have high nutritional value and may be useful in the development of human food and animal feeds.