Contribution of the polymerization of protein by disulfide bonding to increased gel strength of walleye pollack surimi gel with preheating time

ABSTRACT: To clarify the contribution of polymerization of myosin heavy chain (MHC) by disulfide bonding to increased gel strength of cooked gel via preheating, the pastes of walleye pollack surimi (SS and C grades) were preheated at 25°C and 40°C for a variety of hours prior to heating at 80°C for 20 min. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) patterns of cooked gels were analyzed with and without reducing the samples, which were solubilized in 8 M urea–2% SDS solution. The formation of polymers by disulfide bonding in cooked gels was almost constant in each of the SS and C grade surimi gels despite the period of preheating. Therefore, it was suggested that polymerization by disulfide bonding occurred during cooking at 80°C and not during preheating.     

Inhibiting effect of polymerization and degradation of myosin heavy chain during preheating at 30°C and 50°C on the gel-forming ability of walleye pollack surimi

ABSTRACT: To confirm the contribution of polymerization and degradation of myosin heavy chain (MHC) during preheating to the gel-forming ability of fish meat paste, walleye pollack surimi paste was preheated at 30°C and 50°C prior to heating at 80°C in the presence of various inhibitors. At 30°C, ethyleneglycol bis(2-aminoethyl ether) -N,N,N ', N '-tetraacetic acid (EGTA) and ethylenediaminetetraacitic acid (EDTA) inhibited gel formation as well as the polymerization of MHC, whereas dithiothreitol (DTT) and leupeptin promoted gel formation, which was accompanied by the enhancement of MHC polymerization and decreased MHC degradation, respectively. At 50°C, leupeptin inhibited MHC degradation and improved gel strength, whereas EGTA, EDTA and DTT had no effect on MHC polymerization and degradation and did not affect gel formation. The results demonstrate that the gel strength of cooked gel (80°C) is not affected by preheating at 30°C and 50°C and does not inhibit polymerization and degradation. Results suggest that the gel strength of cooked gel is dependent on the polymerization and degradation of MHC during preheating.     

Effect of pH on the gelation of walleye pollack surimi and carp actomyosin pastes

ABSTRACT: The effect of pH on thermal gelation and transglutaminase (TGase; EC2.3.2.13)-induced suwari (setting) of surimi and actomyosin pastes was investigated. A strong and elastic gel was produced from walleye pollack surimi paste at pH 7.0 in the presence of Ca²⁺ using a two-step heating method. In contrast, walleye pollack actomyosin paste formed a weak gel under the same conditions as a result of the low concentration of endogenous TGase. In the presence of EGTA [ethyleneglycol bis(2-aminoethylether) tetraacetic acid], weak gels were formed at pH values of 7.0 and 6.0. Non-proteolytic modori (gel weakening) occurred extensively in the course of actomyosin gelation, but not in surimi gelation. Maximum TGase-induced myosin heavy chain cross-linking was observed at a slightly higher pH of 7.5 than at the optimal pH of endogenous TGase activity; the difference being derived from different substrates. Gelation of carp actomyosin paste at pH values of 5.5, 6.0, 6.5 and 7.0 was monitored by measuring storage modulus (G') and loss modulus (G"). A weak gel was formed at all pH values, but a slightly rigid and less elastic gel was obtained at lower pH values. The addition of microbial TGase (MTGase) formed strong elastic gels at pH 7.0 and 6.5. MTGase cross-linked myosin heavy chains even at pH 5.5, but contributed neither to suwari response nor strong gel formation. Overall, results suggest that the optimal pH for the gelation of surimi paste from easy-setting fish species is a compromise between the pH-optima of TGase activity and of preferable actomyosin conformation for myosin cross-linking.     

Improvement of Cold and Thermally Induced Gelation of Giant Squid (Dosidicus gigas) Surimi

Depending on the season of capture, giant squid (Dosidicus gigas) surimi processed by isoelectric precipitation presents low gel strength. Addition of microbial transglutaminase (MTGase) and application of high isostatic pressure (300 MPa) to improve physicochemical properties were assayed for purposes of making “suwari” gels and heated gels for use in restructured products which have a raw or cooked appearance. The physicochemical properties of both pressurized and unpressurized gels induced by application of 30°C/1 h improved when MTGase was added. In contrast, addition of MTGase was less effective in gels subsequently heated at 90°C/30 min after 30°C/1 h. High pressure treatment for 30 min at 300 MPa and 15°C helped to produce gels with better mechanical and water binding properties, whether treated for 30°C/1 h only or for 30°C/1 h plus 30-min heat treatment at 90°C. High pressure treatment also reduced lightness.     

Study on Gel Properties of Silver Carp (Hypophthalmichthys molitrix) and White Croaker (Argyrosomus argentatus) Blended Surimi at Different Setting Conditions

Gel properties of blends of surimi from silver carp and white croaker set for a range of times and at different temperatures were evaluated for breaking force, breaking distance, and whiteness. Total protein content was 12% in all samples, and the concentration of white croaker surimi protein in the blends was designed at 0, 10, 20, 30, and 40%. The blended surimi had greater gel strength than either of the two types of surimi alone. Breaking force and breaking distance after setting were also significantly higher than that of the blended surimi cooked directly without setting. The blended surimi exhibited less modori than either of the two types of surimi alone heated to 50°C. The whiteness values of the blended surimi were significantly higher than that of white croaker alone. These results indicate there are advantages to be gained in utilizing surimi from freshwater silver carp in blends with surimi from marine captured white croaker which is becoming limited in supply.     

Characterization of fast skeletal myosin from white croaker in comparison with that from walleye pollack

ABSTRACT:   Enzymatic and structural properties of white croaker fast skeletal muscle myosin were determined and compared with those of walleye pollack counterpart. Ca²⁺-ATPase activity of white croaker myosin was decreased to approximately 70% of the original activity during 1 day of storage at 0°C and pH 7.0 in 0.5 M KCl and 0.1 mM dithiothreitol, whereas that of walleye pollack was decreased to approximately 20% under the same condition. The activation energy ( E _a) for inactivation of white croaker myosin calculated by the Arrhenius plot for inactivation rate constant (K_D) was 1.2-fold higher than that of walleye pollack. While Ca²⁺-ATPase showed a similar KCl-dependency for the two species, the maximal activity was observed at pH 6.2 and 6.3 for white croaker and walleye pollack, respectively. Actin-activated myosin Mg²⁺-ATPase activity of white croaker was approximately half that of walleye pollack at 0.05 M KCl and pH 7.0, although the two myosins showed a similar affinity to F-actin with K _m of 1.7 and 1.4, respectively. Limited proteolysis with α-chymotrypsin cleaved heat-denatured white croaker myosin mainly at heavy meromyosin/light meromyosin (HMM/LMM) junction, whereas walleye pollack myosin was cleaved at several sites in LMM as well as at the HMM/LMM junction.     

Gel Forming Ability of Tropical Tilapia Surimi as Compared with Alaska Pollock and Pacific Whiting Surimi

Abstract

Setting and thermal treatment effects on texture and color of tropical tilapia surimi gels were compared to Alaska pollock and Pacific whiting gels. Heat treatments that most favored intrinsic gelling factors of a fish species exhibited strong gel formation. Whiteness values increased as total thermal inputs increased, which reflect the increasing opacity of the gels. Pollock gels were generally the strongest and whitest. Tilapia gel quality was generally second to pollock gels, however, in heat treatments using setting temperatures ≥ 40 °C, tilapia gels were comparable (60 °C setting) or superior (40 °C setting) to pollock gels. The optimum heat treatment for tilapia surimi appeared to be a 40° C setting for 1 hr followed by a 90 °C cook for 15 min. SDS-PAGE patterns of gels prepared with 60°C setting followed by 90 °C cooking elucidated the various degree of protein degradation depending upon the species in a descending order of whiting, tilapia, and pollock.     

Optimum blends of different grades of surimi determined by non-linear programming

SUMMARY: The texture and color properties of surimi gels consisting of pollack surimi, golden threadfin-bream surimi, and low-grade hairtail surimi in various ratios were determined based on a mixture design. Surimi gels were produced by heating at 90°C for 20 min with the addition of 2% NaCl. The texture and color properties of blended surimi from various grades can be represented as non-linear functions. Therefore, non-linear programming was found to be appropriate for determining the optimum formulation for surimi products blended from various grades of surimi. About 3.3% to 18.8% of hairtail surimi could be used when blending with high-grade surimi to produce surimi seafood.     

Effect of Setting Condition on the Gel-Forming Ability of Rohu

To investigate the setting condition of the gel-forming ability of rohu, optimum setting temperature for strong and weak gels of unwashed and washed rohu gel and optimum setting time for maximum proteolytic activity were investigated. Nine setting temperatures were studied for textural properties and trichloroacetic acid (TCA)-soluble peptide contents. Both unwashed gel (UW-gel) and washed gel (W-gel) showed similar optimum setting conditions for producing a strong gel that was set at 40°C for 30 min, followed by heating at 90°C for 20 min. They displayed different optimum setting conditions for weak gel. Weak gel from the degradation of UW-gel and W-gel formed at 65 and 60°C, respectively. The occurrence of protein degradation of W-gels during setting at 60°C suggested that washing did not remove the endogenous protease, and the degradation of unwashed and washed mince was due to water-soluble protease and myofibril-bound protease, respectively. Eight setting times for maximum proteolytic activity were shown by the TCA-soluble peptide contents, accompanying the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) pattern. Both gels had similar results for setting for 120 min.     

Effect of pH-shifting on the gel forming characteristics of salt-ground meat from walleye pollack

ABSTRACT:   In order to elucidate the mechanism of the changes in gel forming characteristics of fish meat by pH-lowering, the gelation-temperature curve and the gelation-moisture content curve were examined using the acidified walleye pollack surimi or neutralized one after acidification. In the gelation-temperature curve, the gel strength was highest at 30°C and lowest at approximately 50–60°C, irrespective of pH shifting. The gel strength at 30°C and 80°C decreased with the decrease in pH value. The neutralization of acidified surimi improved the gel strength, but it was considerably lower than the original gel strength. The gel strength at 50–60°C was not affected by pH lowering. The gel strength at 80°C could not be revived to the original by pH readjustment, either in the presence or in the absence of EDTA. These results suggest that irreversible changes of meat protein take place under the low pH, and the oxidation ability of sulfhydryl (SH) groups of protein molecule is not affected by pH-shifting.     

Oxygen permeability and antioxidative properties of edible surimi films

The oxygen permeability (PO₂) and antioxidative activity of edible surimi films were investigated. Surimi films with lower PO₂ were successfully prepared by heating the film-forming solutions (pH 3) at 70°C for 20 min. On the other hand, it was revealed that surimi films themselves possess antioxidative properties based on 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity and reducing power. Sardine oils in surimi film pouches were stored for 40 days at 40°C in the dark. Both peroxide value and thiobarbituric acid reactive substances values increased slightly during the initial period of storage but decreased significantly at the later stage of storage, indicating that oxidation of sardine oil was effectively hindered by packing in surimi film pouches. Furthermore, polymerization of myosin heavy chain in surimi films was observed during the storage at 40°C.     

TMAOase, trimethylamine-N-oxide demethylase, isa thermostable and active enzyme at 80°C

ABSTRACT:   Purified trimethylamine- N -oxide demethylase (TMAOase)from walleye pollack muscle is a thermostable protein that was notinactivated after heating at 80°C for 30 min.The heated enzyme was electrophoresed in the same manner as fornative enzyme. Circular dichroism (CD) spectra for purified enzymechanged reversibly in the temperature range of 10–80°C.As the enzyme was still active at 80°C, the CD spectralchange did not directly relate to enzyme activity. TMAOase activity inthe myofibrillar fraction decreased sharply above 30°C,but was extracted and recovered from the heated myofibrillar fraction,suggesting that the activity seemed to be interrupted and apparently inactivateddue to the thermal alteration of myofibrillar proteins or some unknownfactors. The complicated profile found in dimethylamine (DMA) formationfrom trimethylamine- N -oxide (TMAO) in walleye pollack muscleduring heating consisted of both enzymic and non-enzymic processes.Most DMA was produced enzymatically below 40°C and interruptedabove 40°C. Therefore, DMA and trimethylamine was formednon-enzymatically at high temperatures regardless of the presenceof native enzyme. A new, simple and easy purification method wasproposed based on the thermostable nature of the enzyme.     

Gel Forming Ability and Other Properties of Eleven Underutilized Tropical Marine Fish Species

Abstract

The gel forming ability and other characteristics of the mince of 11 underutilized marine fish were studied. They were Bombay duck, silverbelly, sea catfish, silver jewfish, jewelled shad, queenfish, Spanish mackerel, hardtail, Indian tuna, tripletail and false conger eel. Mince was prepared from fillet and a portion of the mince was washed two times with cold water (5°C) containing 0.1% NaCl. Both washed and unwashed mince were ground with 3% NaCl. Ground paste was then stuffed into plastic tube and heated for one- and two-step heating. In the one-step heating, the tubes were subjected to 25°, 30°, 35°, 40°, 50°, 60°, 70° and 80°C for 60, 120 and 180 min. In the two-step heating, the tubes were pre-heated at 25°, 30°, 35°, 40°, 50°, 60°, 70° and 80°C for 60, 120 and 180 min. After the pre-heating, the tubes were immediately subjected to 85°C for 30 min. The gel was subjected to puncture, folding, expressible moisture and sensory tests.

Two-step heating distinctly improved the gel strength compared to the one-step heating. The improvement due to two-step heating was more at low preheating temperatures from 25-35°C. Washing improved the texture and color of all of the gels except Bombay duck and decreased the extent of gel-disintegration in silverbelly, queenfish, sea catfish and hardtail. The gels were set optimally at 35°-40°C for most species. Species variation in the disintegration of the gels was observed. Bombay duck mince produced very weak gel. Neither two-step heating nor washing could improve the gel quality of Bombay duck mince. Our data suggested that jewelled shad, queenfish, silver jewfish, sea catfish, tripletail and false conger eel could be suitable as the material for surimi.     

添加圆苞车前子壳粉的鲢鱼糜凝胶工艺优化及其凝胶特性

圆苞车前子壳粉(psyllium husk powder, PHP)是一种富含膳食纤维的食品亲水胶体。为了解其在鱼糜制品中的作用,本实验以冷冻鲢鱼糜为研究对象,以凝胶强度和持水性(water holding capacity, WHC)为考察指标,研究了PHP的添加量、凝胶化温度和凝胶化时间3个因素对鱼糜凝胶特性的影响。在单因素试验的基础上,进行了三因素三水平的正交试验和验证试验。正交试验结果得到最佳工艺条件:PHP添加量0.1%,凝胶化温度45°C,凝胶化时间2 h。单因素试验结果表明,添加适量PHP (0.1%～0.3%)能够增加鱼糜凝胶的硬度和WHC,但对破断距离有不利影响;根据验证试验中蒸煮损失率、SDS-聚丙烯酰胺凝胶电泳(SDS-polyacrylamide gel electrophoresis, SDS-PAGE)和扫描电镜(scanning electron microscopy, SEM)分析,PHP的添加降低了较高凝胶化温度时凝胶的蒸煮损失,PHP或许可以促进肌球蛋白重链(myosin heavy chain, MHC)分子间交联,减缓蛋白质降解,形成更加致密的凝胶结构。本研究对PHP作为一种新型食品原料应用于开发优质健康鱼糜制品进行了初步的探究,以期为丰富亲水胶在影响鱼糜凝胶特性中的应用研究提供一定的参考。     

Effect of Squid Melanin-Free Ink and Pre-Emulsification on Properties and Stability of Surimi Gel Fortified with Seabass Oil during Refrigerated Storage

ABSTRACT

The effects of melanin-free ink (MFI) and pre-emulsification on gel properties and stability of bigeye snapper surimi gel fortified with seabass oil during refrigerated storage of 10 days were studied. Lipid oxidation as determined by peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) of surimi gel increased as the level of seabass oil increased (P < 0.05). When MFI was incorporated into surimi gel, lower PV was obtained throughout the storage (P < 0.05). Addition of seabass oil pre-emulsified with soy protein isolate (SPI) in the presence of MFI yielded surimi gel with the highest breaking force and could improve oxidative stability during refrigerated storage (P < 0.05). Slight decrease in whiteness was found in surimi gel added with MFI, while those added with MFI along with pre-emulsified seabass oil showed increased whiteness (P < 0.05). Addition of MFI did not affect total viable count and psychrophilic bacterial count in surimi gels. Thus, the incorporation of pre-emulsified seabass oil prepared using SPI in conjunction with MFI could improve quality and oxidative stability of gel from bigeye snapper surimi.     

Identification of the glutamine residue that may be involved in the transglutaminase-mediated intramolecular crosslinking of carp and walleye pollack myosin

In order to elucidate the molecular mechanism of transglutaminase-mediated myosin crosslinking, a fluorescent monodansylcadaverine (MDC) was incorporated into carp Cyprinus carpio myosin and the reactive Gln residues were analyzed by cyanogen bromide cleavage. The fluorescence was predominantly detected in a 10.5 kDa BrCN fragment, assumed to be located in subfragment 2 of the myosin heavy chain. Furthermore, lysyl endopeptidase digestion of the 10.5 kDa fragment revealed that MDC was specifically incorporated into the 520th Gln residue of the subfragment 2 domain. When meat paste prepared from frozen walleye pollack (Theragra chalcogramma) surimi was incubated with MDC, the fluorescence was mostly observed in a 16 kDa BrCN fragment and also slightly detected in other three bands. By digesting the 16 kDa fragment with lysyl endopeptidase, it was elucidated that MDC was incorporated specifically into Gln-520 of myosin subfragment 2, as also detected in carp. This domain around Gln-520 is likely to be a critical region for the formation of myosin heavy chain dimers that both fish species have in common. In walleye pollack, other reactive Gln residues are presumed to exist at the C-terminus of the light meromyosin. This slight difference may have a significant effect on the capacity of myosin to form tetramers or even larger multimers.     

Comparative Properties of Bluefish (Pomatomus saltatrix) Gels Formulated by High Hydrostatic Pressure and Heat

High pressure was applied at levels of 300 to 3,742 atm for 30 min to formulate gels from bluefish meat paste, and the properties of the resulting gels were compared with those of heat-induced gels formulated at 90°C for 20 min, or at 60°C for 60 min. The moisture content of the pressure-induced gels was similar to that of the heat-induced gels, while protein contents and pH values of pressurized gels were slightly lower than those formulated by heat. Gels formed by pressure were more translucent as compared with those formulated by heat. Texture measurement indicated that there were no significant differences between the elasticities of the gels obtained under various pressures, although gel firmness increased with pressure. Overall, the heat-induced gels formulated at 90°C for 20 min were firmer but had similar elasticities to pressure-induced gels, while gels formulated at 60°C for 60 min were comparatively softer and had lower elasticities. The salt-extractable protein and protein digestibility studies indicated that pressure treatment formed gels with less protein denaturation and which were more digestible than the fish gels formulated by heat. The results from proteolytic activity studies showed that the pressure range used in this study was less effective in inactivating the endogenous proteases in the fish flesh than heat.     

Effect of setting conditions on mechanical properties of acid-induced Kamaboko gel from squid <Emphasis Type="Italic">Todarodes pacificus</Emphasis> mantle muscle meat

Squid Todarodes pacificus suwari gels, set at various temperatures and times, and acid-induced kamaboko gel, which was prepared by soaking suwari gel in 5% acetic acid for 20 h, were studied to evaluate the mechanical properties that are affected by setting conditions. Unset squid meat paste did not form a gel when soaked in acetic acid. The breaking strength of both suwari gel and acid-induced kamaboko gel showed a tendency to increase with setting temperature and time. SDS-PAGE analysis of suwari gel and acid-induced kamaboko gel, which were set at various temperatures and times, showed that myosin heavy chain (MHC) was observed at 30°C only for the first hour. The intensity of the MHC band at 30°C gradually decreased with setting time, while the intensity of the polymer band gradually increased with setting time. These results suggest that the protein-protein bonds in suwari gel affect the final texture of acid-induced kamaboko gel. Based on the analysis of the mechanical properties, and in consideration of the fact that the purpose of this experiment was to reduce energy usage, the best setting condition was determined to be 40°C for 3 h.     

Gelation Characteristics of Mince and Washed Mince From Small-Scale Mud Carp and Common Carp

Small-scale mud carp (SC, Cirrhina microlepis) and common carp (CC, Cyprinus carpio) are underutilized freshwater fish. The objective of this study was to elucidate the gel-forming ability of mince and washed mince from these species in relation to washing cycles and CaCl₂ addition. Protein loss of up to 67% was observed after three-washing cycles of both species, rendering relatively low yield. About 85–90% of total fat was removed from SC flesh after three-washing cycles, but only two-washing cycles were sufficient for fat removal in CC mince. Mince and washed mince of both species did not exhibit severe proteolysis with low autolytic activity at 65°C. SC exhibited superior gel-forming ability to CC. Two- and three-washing cycles in conjunction with a 40°C-preincubation resulted in the highest textural properties of SC and CC, respectively. CaCl₂ increased the breaking force of CC and SC mince gels up to 65 and 95% at 0.3 and 0.5% CaCl₂, respectively, as compared to without CaCl₂. However, it showed negative effects on surimi gels of both species. Both SC and CC are potential freshwater fish resources for mince and washed mince production with reasonable good gel-forming ability.     

Antioxidant Properties of Scomber japonicus Hydrolysates Prepared by Enzymatic Hydrolysis

The antioxidant properties of the Pacific chub mackerel (Scomber japonicus) muscle protein hydrolysates prepared by enzymatic hydrolysis were investigated. After enzyme hydrolysis at 50°C for 60 min, more than 80% of the S. japonicus muscle protein was hydrolyzed. The highest 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity (71.69%) occurred in whole muscle protein hydrolysates treated at 50°C for 30 min with Protamex, and the highest 2,2?-azino-bis(3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) radical-scavenging activity (95.39%) was observed in white muscle protein hydrolysates treated at 50°C for 30 min with Neutrase. The highest superoxide dismutase (SOD)-like activity (32.84%) was recorded in white muscle protein hydrolysates treated at 50°C for 120 min with Protamex. Changes in the molecular weight distribution of S. japonicus muscle proteins after enzymatic hydrolysis were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A robust and a convenient enzyme hydrolysis technique for obtaining S. japonicus muscle protein hydrolysates with useful biological activities, within a short time (<2 h) is proposed.