Protease Inhibitor Effects on Torsion Measurements and Autolysis of Pacific Whiting Surimi

Beef plasma protein (BPP), egg white and potato extract were tested for their ability to inhibit proteolysis in fish mince and surimi made from Pacific whiting (Merluccius productus). Strong inhibition resulted from all three compounds in fish mince when measured by autolysis. However, when tested in surimi significant differences occurred among the compounds. BPP showed strongest inhibition of proteolytic effect followed by egg white and potato extract when measured by autolysis, gel electrophoresis and torsion. BPP was an effective inhibitor in surimi at a concentration as low as 1%.     

Development of a healthy low-fat fish sausage containing dietary fibre

In order to develop a healthy low-fat fish sausage containing dietary fibre (DF), three fundamental changes were made to an ordinary pork sausage recipe: incorporation of 4% (w/w) of Swelite^® (a DF obtained from inner pea), different levels of pork meat replacement (0%, 50% and 100%) by hake mince and the combination of varying amounts of Fibruline^® (a DF obtained from chicory root) and hake mince (Fibruline: additional hake mince, 2.6:5.2, 5.2:2.6 and 7.8:0.0, % w/w) as a substitute for pork fat. It was found that the addition of Swelite to pork sausage favoured greater gel strength and hardness. On the contrary, increasing levels of pork meat replacement by hake reduced the sausages’ gel strength and hardness. Finally, sausages without pork fat showed promising textural and colour parameters. High-Fibruline sausages were less cohesive and chewable than pork fat sausages (control), but also exhibited a greater gel strength. Low-Fibruline ones presented almost all textural properties similar to the control, with exception of hardness and gumminess. Therefore, regarding some key textural parameters, it was possible to produce low-fat fish sausages similar to the ordinary pork sausages.     

Alaska Pollock (Theragra chalcogramma) Mince and Surimi as Partial Meat Substitutes in Frankfurters: N-Nitrosodimethylamine Formation

Studies were conducted to determine the feasibility of using 3 forms of Alaska pollock (AP) as a partial substitute for meat in a nitrite-cured product. The effect of several pre- and post-processing storage conditions on N-nitrosodimethylamine (NDMA) were studied in frankfurters substituted with 15 and 50% unwashed and washed mince and surimi. In franks made with unwashed mince, frozen stored prior to processing, NDMA increased during frozen storage; little change was observed in franks containing washed mince and surimi. Refrigerated storage of franks generally showed a decrease in NDMA with all 3 forms of fish. Overall, the use of washed mince and surimi at the 15% substitution level gave mean values less than 2 ppb NDMA, even after boiling. The results indicate that 50% unwashed AP mince should not be used as a partial substitute for meat in frankfurters.     

Characteristics of surimi and kamaboko from sardines

Sardines of varying freshness (1 to 3 days in ice) were manually or mechanically processed into fish mince and surimi using 1 to 3 washing steps. Standard kamaboko gels were prepared by grinding thawed surimi with 3% NaCl and 5% potato starch, stuffing into sausage casings, holding at 37°C for 30 min and cooking at 90°C for 50 min.
Three washing steps decreased the yield of washed fish mince (21-27 g per 100 g of whole fish), and the protein recovery (50-55% of that present in the unwashed mince), but caused efficient lipid removal (80%) leading to surimi containing only 0.2-1.4% lipid. The texture and colour of the final kamaboko were also improved.
The texture parameters (folding score, rigidity, elasticity index and gel strength) of kamaboko prepared from very fresh sardines were markedly enhanced by gel setting during incubation at 37°C for 30 min or 4°C for 24 hr. Incubation at 60°C for 30 min led to soft, coarse gels. Omitting potato starch or replacement by spray dried egg white had little effect on texture.
The texture of kamaboko prepared from less fresh sardines was less firm and elastic, did not improve when incubation was carried out at 37 or 4°C before cooking, and was softer and coarser after incubation at 60°C. Partial or total replacement of potato starch by egg white, soy protein isolate or bovine serum albumin markedly improved the texture. Egg white also increased the colour lightness.     

Electrical Properties of Fish Mince During Multi-frequency Ohmic Heating

A multi-frequency ohmic heating system with 30 Hz～1 MHz range which could deliver 250 watts was developed for measuring electrical conductivity and absolute dielectric loss of food samples. Pacific whiting surimi paste and stabilized mince in the 20～70°C range were tested at frequencies from 55 Hz to 200 kHz. Sample impedance decreased slightly with frequency. The DC electrical conductivity (σ_dc) and absolute dielectric loss (ε″) of Pacific whiting surimi paste increased with temperature and salt concentration; adc and ε″ of the stabilized mince increased with temperature. Empirical models of electrical properties for surimi paste (moisture content 79% and salt at 1, 2 or 3%) and stabilized mince (77% moisture and 0.74% salt) were derived. Electrolytic corrosion diminished with frequency.     

Thermal Transitions of Actomyosin and Surimi Prepared from Atlantic Croaker as Studied by Differential Scanning Calorimetry

Differential scanning calorimetry (DSC) was used to investigate thermal transitions of fish mince (surimi) and actomyosin from croaker. Three endothermic peaks were observed in DSC thermograms of surimi. After addition of salt, transition temperatures shifted to lower temperatures. Preheating samples containing 3% salt at various temperatures showed that 40°C heating caused the first peak to disappear, and preheating at temperatures higher than 50°C caused virtual disappearance of all transition peaks. Low temperature storage (4°C) of samples caused no significant change in thermograms of salted or unsalted surimi over a 5-day storage period. Evidence suggests that changes of fish protein during low temperature “setting” are different from those occurring during high temperature “setting.”     

Comparison of the properties of gels derived from cod surimi and from unwashed and once-washed cod mince

Surimi is a washed and stabilized fish mince with valuable gel-forming properties. However the preparation process is wasteful and a modified process, with less washing, was investigated. The properties of the gels (cooked at 40°C for 30 min followed by 90°C for 40 min) were assessed by instrumental and sensory methods. Gels prepared from surimi were soft and very flexible; gels from unwashed fresh cod mince were firmer and less elastic, but still gave a maximum score on the folding test. A single wash made the gels firmer and slightly more elastic than those from unwashed mince. Gels from unwashed mince of frozen cod were firmer than surimi-derived gels, and were the most brittle. Sensory assessment of the gels found no difference in firmness between the different washing procedures and small differences in elasticity and toughness. The addition of starch (fish: starch, 100:5, w/w) increased the firmness and sensory toughness of each derived gel, and reduced the elasticity in the mouth of non-surimi gels. The colour of gels derived directly from fish was off-white, compared with the white surimi-derived gels. The colour of the former darkened during frozen storage, but lightened again after thawing.     

Identification by GeLC‐MS/MS of Trypsin Inhibitor in Sarcoplasmic Proteins of Three Tropical Fish and Characterization of Their Inhibitory Properties

Sarcoplasmic proteins from 3 fish species were fractionated by 50% to 70% ammonium sulfate precipitation. Lyophilized fractionated sarcoplasmic proteins of threadfin bream (TB‐SP), bigeye snapper (BS‐SP), and yellow croaker (YC‐SP) showed 80% to 92% trypsin inhibitory activity. Trypsin inhibitory activity staining gel electrophoresis revealed bands at 32, 33, 37, 45, 48, and 50 kDa for the 3 species, and a band at 95 kDa was observed for TB‐SP and YC‐SP. Alpha‐1‐antitrypsin with molecular mass of 45 to 50 kDa was identified in YC‐SP by gel‐based liquid chromatography‐tandem mass spectrometry (GeLC‐MS/MS). Other major protein bands appeared on trypsin activity staining included phosphorylase, glyceraldehyde‐3‐phosphate dehydrogenase, and creatine kinase with molecular mass of 95 and 35 to 40 kDa, respectively. But, these 3 proteins did not show true trypsin inhibitory activity. Trypsin inhibitory activity of fractionated sarcoplasmic proteins showed good stability, with >80% activity retained at 60 °C and up to 0.6 M NaCl. TB‐SP showed the highest inhibitory activity against autolysis of washed threadfin bream mince at 65 °C. Addition of 0.5% or 1% TB‐SP improved textural properties of threadfin bream surimi gels preincubated at 37 or 65 °C followed by heating at 90 °C. Therefore, TB‐SP could be a promising protein ingredient for enhancing surimi gel texture.     

Development of lipid oxidation during manufacturing of horse mackerel surimi

When fatty fish are transformed into surimi, lipid oxidation takes place, decreasing the quality of the product. This study was aimed to identify the critical stages of the process in terms of the development of lipid oxidation. Horse mackerels were transformed into surimi on a pilot line and samples taken (hand‐skinned fillets = minced fillets, mince, washed and refined minces, paste, surimi and washing water). Most of the lipids were removed during the process and neutral lipids were lost in higher proportion than polar lipids. As a consequence, total lipids of surimi contained more polyunsaturated fatty acids (338 ± 19 g kg^?1) than total lipids of the minced fillets (220 ± 8 g kg^?1). Thiobarbituric acid reactive substances (TBARS) was higher in the minced fillets than in the mince because less subcutaneous fat and dark muscle were removed during hand‐mincing, indicating that the settings of the skinning–deboning machine can strongly influence the final quality of the product. Concentrations of lipid oxidation products increased significantly during the next stages of surimi processing. The increase was more pronounced for TBARS than hydroperoxides. Concentrations in hydroperoxides were similar in mince and washed mince (15.3 ± 2.8 and 16.6 ± 2.8 mmoles kg^?1 lipid) and increased in refined mince (29.6 ± 2.8 mmoles kg^?1 lipid). TBARS accounted for 2.7 ± 1.0 mg kg^?1 lipid in mince, 40.4 ± 2.3 mg kg^?1 lipid in washed mince and 237 ± 7 mg kg^?1 lipid in refined mince. Hydroperoxides and TBARS were found in appreciable amounts in washing water (76.9 ± 4.7 mmoles kg^?1 lipid and 479 ± 8 mg kg^?1 lipid respectively), when they decreased in surimi (27.3 ± 3.8 mmoles kg^?1 lipid and 44.2 ± 0.8 mg kg^?1 lipid respectively) compared with refined mince. This shows that the last dewatering stage is crucial to ensure surimi quality. Copyright © 2005 Society of Chemical Industry     

Cryoprotective Effects of Some Materials on Cod-Surimi Proteins during Frozen Storage

Freeze-induced protein denaturation of cod surimi was studied as affected by carbohydrates (sucrose and glucose syrup at 8% w/w), polyols, (sorbitol and glycerol at 8% w/w), protein hydrolysates (fish protein and casein hydrolysates at 4% w/w), hydrocolloids (pectin-1% w/w, sodium alginate, lambda- and iota-carrageenan-0.5% w/w) and combinations of the above, (sucrose/sorbitol 1:1 mixture at 8% w/w, or combined with protein hydrolysates at 4% w/w). Salt extract-able protein (SEP) and heat induced denaturation by differential by differential scanning calorimetry (DSC) were used to monitor protein changes in surimi stored 16 wk at -20°C. The best cryoprotection effect was achieved from sorbitol, glucose syrup (DE = 60), sucrose and sucrose/sorbitol 1:1 w/w mixture at 8% w/w in surimi. Correlations between certain DSC parameters and SEP were high.     

Heat-Induced Gelation Properties of Surimi From Mechanically Separated Chicken

Chicken surimi was prepared from fresh mechanically separated chicken meat using a sodium bicarbonate washing process. The heat-induced gelation properties were assessed under different conditions of pH, temperature, heating rate, protein and sodium tripolyphosphate (TPP) concentrations. Surimi gel strength increased (p < 0.05) after: reducing pH from 6.4 to 6.0, increasing temperature from 40°C to 80°C, reducing heating rate from 5°C/min to 1°C/min, increasing protein concentration from 4% (w/w) to 8% (w/w) or addition of 0.3% (w/w) TPP. Freeze thaw stability studies revealed that the gel strength of surimi decreased (p < 0.05) when subjected to frozen storage at – 18°C.     

PROTEIN INSOLUBILIZATION IN FROZEN GREENLAND HALIBUT (REINHARDTIUS HIPPOGLOSSOIDES)

Frozen storage of minced Greenland halibut (Reinhardtius hippoglossoides) at -10°C resulted in a rapid loss in salt solubility of “my ofibrillar proteins” (approximately 50% in 15 days) and in a gradual loss in water solubility of “sarcoplasmic proteins” (approximately 40% in 120 days). The water and salt inextractable protein from frozen mince (R) was completely soluble in 4% sodium dodecylsulfate (SDS) when a disulfide bond reducing agent such as mercaptoethanol (ME) was present. Other reagents, including urea and Triton X 100, were less effective in solubilizing the protein from mince after frozen storage. Evidence presented supports the thesis that disulfide bond formation contributes to the observed loss in protein extractability during frozen storage of mince. Addition of various thiol reagents to mince prior to freezing was effective in minimizing protein insolubilization. An estimated 50% of the reduced cysteine associated with protein is oxidized in mince after long time storage at -10°C. However, the kinetics of disulfide bond formation do not parallel the time course of protein insolubilization; accordingly, the possibility that disulfide bond formation is a secondary event cannot be excluded. Other lines of evidence indicate that additional covalent or strong bond interactions contribute to the formation of(R). Solubilization and boiling of(R) in SDS/ME does not dissociate high molecular weight aggregates (500–1000 Kdaltons) although these aggregates are disrupted by sodium borohydride reduction. In addition, the degree of hydrolysis of proteins in frozen mince by certain proteolytic enzymes is lower then that of fresh mince despite the complete solubilization of the mince protein by these proteolytic enzymes. The mince protein from frozen fish contains substantially more fluorescence than that of fresh fish. Fluorescence is associated with the high molecular weight peptide fraction after pepsin catalyzed hydrolysis. The amino acid composition of mince proteins after HCl or formic acid/HCl hydrolysis did not change appreciably as a result of frozen storage. The data indicate that covalent bond formation by sulfhydryl residues and other borohydride and acid labile linkages contribute to the loss in solubility of the protein in Greenland halibut mince during frozen storage at -10°C.     

Characteristics of mince from pond-bred silvercarp (Hypophthalmichthys molitrix) and preliminary experiments on its use in sausages

The quality of fresh, chilled and frozen mince made from the flesh of pond-bred silvercarp (Hypophthalmichthys molitrix (Val)) was evaluated. Flavour panel scores for all minces were acceptable and this quality did not deteriorate during storage for one week in a domestic refrigerator (5–6°C) or for one year at ?20°C. The levels of oxidative rancidity (2-thiobarbituric acid analysis) in the minces were consistent with this maintenance of quality. Aerobic plate counts at 25°C of fresh mince rose from 7–9 × 10⁴ to 1.1 × 10⁵ after 6 days' storage at 5–6°C. Coliform and psychrotroph counts rose from 1.0 × 10¹ and 7–8 × 10³ to 5–4 × 10² and 1.3 × 10⁴, respectively. The functional and textural properties of pond-bred silvercarp mince in terms of salt-soluble protein content, water binding capacity and penetrometer values were measured on fresh, refrigerated and frozen samples. All-fish sausages and frankfurters developed from pond-bred silvercarp mince were assessed in laboratory and consumer tests against commercial beef sausages and frankfurters. The fish products competed well and achieved levels of acceptance similar to those of the beef-containing products. Texture measurements revealed slight changes in the fish products during storage for 7 days at 5–6°C. No spoilage was detected in any of the products after a week at 5–6°C. The nutritional advantages of the fish sausages and frankfurters are discussed.     

Gelation of Beef Heart Surimi as Affected by Antioxidants

Oxidation of proteins (carbonyls) and lipids (TBARS) in beef heart surimi-like materials during preparation and storage (2°C) was inhibited by propyl gallate (0.02%) or α-tocopherol (0.2%). Inhibition of oxidation did not affect surimi gel property (storage modulus, G'). Storage promoted oxidation of proteins in 0.2% ascorbate-washed mince, leading to increases in peak (～ 55°C) and final (70°C) G' of thermally induced surimi gel. Protein carbonyls in stored surimi and its sol (salted), as well as TBARS of the sol, strongly correlated with both peak and final G of gels. Incorporation of tripolyphosphate into washed mince promoted gelation whether the surimi-like material was oxidized or not.     

CHANGES IN PROXIMATE COMPOSITION OF THORNBACK RAY (RAJA CLAVATA, L. 1758) SURIMI DURING WASHING AND FROZEN STORAGE

The effects of frozen storage of surimi produced from thornback ray and the washing of mince on the chemical composition were investigated. The crude ash content which was initially found as 1.38% in raw thornback ray decreased approximately 12 and 80% after the first and second washing, respectively. After the third washing, crude ash content increased to 207% of the amount in the second washing because of addition of salt to the last washing water. The crude protein content of mince also decreased approximately to 28 and 20% after the first and second washing, respectively. After the third washing, the decrease in the lipid levels was approximately 30%. At the end of 6 months of frozen storage at − 23.8  ±  2C, dry matter, crude ash and crude protein contents increased in a greater ratio in surimi containing 4% sorbitol, 4% sucrose and 0.3% Na-tripolyphosphate than surimi prepared with 8% sorbitol and 0.3% Na-tripolyphosphate.

PRACTICAL APPLICATIONS

In the present study, the effects of frozen storage of surimi produced from thornback ray and the washing of mince on the chemical composition were investigated. Washing procedure significantly decreased crude ash, crude protein and crude fat content of mince. There were significant differences in moisture, crude ash, and crude protein contents during the 6 months storage period of frozen surimi obtained by using different cryoprotectant mixtures. The folding test scores were highest in fresh surimi and during the first two months of storage. Thornback ray can be used for the production of surimi.     

Effects of Hydrolysates from Silver Carp (Hypophthalmichthys molitrix) Scales on Rancidity Stability and Gel Properties of Fish Products

This study aimed to evaluate the effectiveness of hydrolysates, which were obtained from the scales of silver carp (Hypophthalmichthys molitrix) by papain, flavourzyme, and Alcalase 2.4 L, as natural antioxidants in silver carp mince and surimi gels during storage at 4 °C. The hydrolysates that possess greater in vitro antioxidant activities (DPPH radical-scavenging activity, Fe²⁺-chelating activity, and reducing power), including hydrolysates catalyzed by papain at 10 min (HP), flavourzyme at 5 min (HF), and Alcalase 2.4 L at 5 min (HA), were chosen as additives. Color, cooking loss, conjugated dienes (CDs), thiobarbituric acid reactive substances (TBARS), fatty acids, and sensory scores of mince were measured on days 0, 2, 4, 6, and 8 during 4 °C storage; additionally, whiteness, breaking force, deformation, gel strength, and sensory score of surimi gels were measured on days 1, 3, 5, 7, 9, and 11 during 4 °C storage. The results indicate that HA was conducive to lowering the cooking loss of mince and that HF significantly (P?<?0.05) reduced the CDs value of mince. For surimi gels, HF improved whiteness, deformation, and gel strength. Hence, HF could serve as a natural antioxidant during early oxidation and improve gel formation of silver carp products.     

The effect of low- or non-sweet additives on the stability of protein functional properties of frozen cod surimi

The cryoprotective effect of low- or non-sweet additives, Palatinit^R, Polydextrose^R, casein hydrolysate and fish protein hydrolysate (at 8% w/w) as well as lactitol (at 4% and 8% w/w) were compared to an industrial control containing sucrose/sorbitol (8% w/w) and a control without additive in cod surimi stored at –20°C for 4 months. Freeze-induced protein denaturation was evaluated monthly by salt extractable protein and differential scanning calorimetry analyses of surimi followed by texture and expressible moisture analyses of surimi cooked gels. Results revealed that protein functionality was similarly maintained during frozen storage by lactitol, Palatinit^R and Polydextrose^R when incorporated in cod surimi at the 8% level. Surimi gels could be produced with textural attributes comparable to 8% w/w sucrose/sorbitol surimi gels, and the level of lactitol in cod surimi could be reduced to 4% w/w without significant alteration of cryoprotection. Some benefits in gel-forming properties were obtained by adding 8% casein hydrolysate to cod surimi.     

Effect of Frozen Storage on the Gel-forming Properties of Hoki (Macruronus novaezelandiae)

The possibility of using frozen hoki to make surimi was investigated. At intervals, fish were thawed under controlled conditions and gels were then made from minces of the flesh. Both puncture and torsion tests showed the quality of gels declined with duration of storage of frozen hoki. This decline was matched by a decrease in pH and an increase in formaldehyde concentration in the frozen flesh. The data suggest a land-based surimi plant could not be operated outside the harvesting season to any appreciable extent using frozen hoki stored at -29°C.     

Comparison of the Thermostability of Red Hake and Alaska Pollock Surimi during Processing

Thermostability of red hake (Vrophycis chuss) mince and its temperature-dependent gel-forming properties were determined while using Alaska pollock (Theragra chalcogrumma) for comparison. Fish mince and surimi were subjected to various washwater, chopping and setting/ cooking temperatures, cooking times at varying salt concentrations and moisture levels. The optimal temperatures for washing and chopping were 15°C and 12°C for red hake and 10°C and 4°C for pollock, respectively. All treatments significantly affected gel properties. For red hake gels, 77% moisture, 2.0% salt, and a 40°C preheat-setting temperature produced the most cohesive gel. Gels of both red hake and pollock gradually became less cohesive with extended cooking time. The results suggest that red hake is more thermally stable than pollock.     

Hydrolysis of surimi wastewater for production of transglutaminase by Enterobacter sp. C2361 and Providencia sp. C1112

Surimi wastewater (SWW) is an industrial wastewater, released during the washing step of surimi preparation from minced fish, that causes environmental problem. In this study, SWW produced from ornate threadfin bream (Nemipterus hexodon) was hydrolysed and used to cultivate Enterobacter sp. C2361 and Providencia sp. C1112 for the production of microbial transglutaminase (MTGase, EC 2.3.2.13). The SWW was repeatedly used to wash the fish mince that gained a final protein content of 3.20% (w/v). The commercial protease, Delvolase was the most appropriate protease used to produce fish protein hydrolysate (FPH) from SWW. The FPH at 40% degree of hydrolysis was used instead of a peptone portion in the SPY medium (3.0% starch, 2.0% peptone, 0.2% yeast extract, 0.2% MgSO(4), 0.2% K(2)HPO(4) and 0.2% KH(2)HPO(4), pH 7.0) to cultivate the tested strains at 37°C, shaking speed at 150rpm. Providencia sp. C1112 produced higher MTGase activity (1.78±0.05U/ml) than Streptoverticillium mobaraense (1.61±0.02U/ml) at 18h of cultivation in FPH medium. On the other hand, the Enterobacter sp. C2361 produced lower MTGase activity (1.18±0.03U/ml).