Gel Strength Enhancement by Addition of Microbial Transglutaminase during Onshore Surimi Manufacture

Surimi from Alaska pollock flesh was manufactured onshore with Microbial transglutaminase (MTGase). Effect of MTGase was investigated by evaluating breaking strength and deformation of gels from MTGase-treated surimi with and without setting at 30°C. Quantitative analysis of ε-(γ-glutamyl)lysine (GL) crosslink was also carried out to monitor the MTGase reaction. In set gels, breaking strength and GL crosslink increased, and myosin heavy chain decreased correspondingly with MTGase concentration. These changes were smaller in gels prepared without setting. Results suggest that surimi gel could be improved through the formation of GL crosslinks by added MTGase in surimi.     

Transglutaminase Effects on Low Temperature Gelation of Fish Protein Sols

Myosin polymerization and formation of ?-(γ-glutamyl)lysine linkages were quantified in Alaska pollock surimi gels which contained no additive (control), or a commercial microbial transglutaminase (MTGase). As preincubation (“setting”) time at 25°C was increased, the gel strength of control and 0.2% MTGase-added samples increased, with greater increases at higher MTGase levels. SDS-PAGE and HPLC analyses showed increasing nondisulfide polymerization and ?-(γ-glutamyl)lysine dipeptide content, with increasing setting time and/or added MTGase. Content of ?-(γ-glutamyl)lysine dipeptide correlated with gel strength (shear stress) and shear modulus at failure (G_f) for these gels. Higher stresses were measured in samples containing 0.2% MTGase than in controls at corresponding levels of ?-(γ-glutamyl)lysine dipeptide, indicating that rate of myosin polymerization may affect ultimate gel strength.     

Suppression of Surimi Gel Setting by Transglutaminase Inhibitors

Three types of salted meat paste (3% NaCl, 3% NaCl plus 0.66% NH₄Cl or 3% NaCl plus 0.2% EDTA) were prepared from high and second grade surimi, set at 30°C up to 4 br, and subsequently heated at 85°C for 30 min. The gel strength, crosslinking of myosin heavy chain (MHC) and ?-(γ-glutamyl)lysine (?-(γ-Glu)Lys) content were determined. With extended setting time, gel strength, crosslinking of MHC and the content of a crosslinked product, ?-(γ-Glu)Lys, increased markedly in the gel from the high grade surimi. Such changes were suppressed considerably in the presence of NH₄Cl and EDTA and were not observed in the gel prepared from second grade surimi. These results indicated an active participation of intrinsic transglutaminase in the setting process.     

NONDISULFIDE COVALENT CROSS-LINKING OF MYOSIN HEAVY CHAIN IN “SETTING” OF ALASKA POLLOCK AND ATLANTIC CROAKER SURIMI1

ABSTRACT Myosin heavy chain (MHC) content of cooked gels of pollock and croaker surimi decreased during preincubation (“setting”) at temperatures ranging from 4–50C. Decreases in MHC content were attributed to either nondisulfide covalent cross-linking or proteolysis. Depending upon which process dominated at a given temperature, formation of stronger or weaker gels occurred, respectively. Maximum production of cross-linked polymers occurred at the optimum setting temperatures, i.e., at 25C for pollock surimi and 40C for croaker surimi. Subsequent cooking of these set gels at 90C decreased the amount of cross-linked polymers formed at the optimum setting temperature. Addition of free lysine-HCl inhibited formation of cross-linked polymers of MHC during setting and the increase in cooked gel strength for both species. This supports published evidence that cross-linking of MHC during setting may be of the ε-amino-(γ-glutamyl) lysine I type, mediated by a transglutaminase enzyme.     

Strength of Protein Gels Prepared with Microbial Transglutaminase as Related to Reaction Conditions

Influence of gelling reaction conditions on the strength of several protein gels prepared with microbial transglutaminase (TGase) was investigated. A method was developed to gel proteins and measure gel breaking strength in a micro well plate. Enzyme concentration range for maximum gel breaking strength varied from 10 to 40 units/g protein. Maxima gel breaking strengths were achieved at 50°C for SPI, caseinate and gelatin and 65°C for egg yolk and egg white proteins. Optimum pH resulting in strong gels was pH 9 for SPI, caseinate, and egg yolk, and pH 6 for gelatin and egg white. Adjusting pH was promoted in egg white the formation of ?-(γ-glutamyl)lysine crosslinks and increased its gel breaking strength.     

ε-(γ-Glutamyl)lysine Crosslink Distribution in Foods as Determined by Improved Method

Quantitative analysis of ε-(γ-glutamyl)lysine crosslink in 127 foods was achieved with a, preliminary separation by reverse phase-HPLC before o-phthalaldehyde derivatization to remove interfering peaks. ε-(γ-Glutamyl)lysine was detected in 96 foods and its contents ranged from 0.2 to 135 μmol/lOOg protein. High levels were found in fish paste products, processed fish, shellfish, meats and soybeans, and raw poultry organs. For fish and meats, the level of ε-(γ-glutamyl)lysine in processed foods and fish paste was relatively higher than that in raw materials. The improved procedure could be applied for screening materials with transglutaminase activities.     

Formation of ε-(γ-Glutamyl) Lysine Cross-Link in Cured Horse Mackerel Meat Induced by Drying

Block meat cut from horse mackerel was cured in 3M NaCl (pH 7.5) in the presence and absence of 5 mM EDTA for 3 hr at 4°C. During drying at 30°C, changes in the myosin heavy chain and ε-(γ-glutamyl)lysine isopeptide bonds in the meat were observed. After 20 hr drying, cross-linking of the myosin heavy chain was observed and ε-(γ-glutamyl)lysine bonds increased eight fold. These changes were inhibited in the presence of EDTA. These results suggested that trans-glutaminase was probably involved in the cross-linking reaction of the myosin heavy chain in the manufacture of dried fish (“Himono” in Japan).     

CHITOSAN AFFECTS TRANSGLUTAMINASE-INDUCED SURIMI GELATION

Effect of chitosan on barred garfish (Hemiramphus far) surimi gel was studied in the presence of EDTA and microbial transglutaminase (MTGase). An increase in breaking force of surimi gels added with 1.0% prawn shell chitosan indicated the gel enhancing effect of chitosan on the heat‐induced gelation of fish myofibrillar proteins. However, gel‐forming ability of surimi containing chitosan was inhibited in the presence of EDTA, especially at higher concentration. Therefore, the enhancing effect of chitosan was possibly mediated through the action of endogenous transglutaminase (TGase) during setting, resulting in the formation of protein‐protein and protein‐chitosan conjugates. In general, addition of MTGase remarkably increased both breaking force and deformation of surimi gel (P<0.05). However, enhancing effect of MTGase was retarded in the presence of chitosan, resulting in lower magnitude of breaking force and deformation (P<0.05). Scanning electron microscopy showed that chitosan particles were uniformly dispersed in the gel matrix. A tightly associated gel network was formed in surimi containing MTGase, whereas a large number of voids were noted in gels with EDTA. These results suggest that chitosan acted as a surimi gel enhancer in combination with endogenous TGase in fish muscle, but hindered gel formation in the presence of MTGase.     

微生物转谷氨酰胺酶诱导下鲢鱼糜凝胶的结构演化规律

以鲢鱼糜为对象,通过检测微生物转谷氨酰胺酶（microbial transglutaminase,MTGase）诱导下不同凝胶化时间形成的鱼糜凝胶的质地特性、交联程度及网络微观结构,探讨鲢鱼糜凝胶的结构演化规律。力学特性结果表明,未添加MTGase（对照组）的鱼糜凝胶的破断力、凹陷深度、硬度、咀嚼性等随凝胶化时间延长显著增大（P＜0.05）,破断力在3～6 h达到平衡（约1 100 g）,凹陷深度在1～2 h达到最大值（约17 mm）,随着凝胶时间延长呈下降趋势;添加MTGase的鱼糜凝胶破断力在3 h时就达到最大值（P＜0.05）,硬度和咀嚼性随时间增加到3 h后趋于平衡,凹陷深度随时间延长逐渐降低（P＜0.05）。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodiumdodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE）结果显示,鲢鱼糜凝胶中肌球蛋白重链含量随凝胶化时间延长显著下降,添加MTGase组肌球蛋白重链含量明显低于对照组。两组鱼糜凝胶的网络孔隙当量直径均随凝胶化时间延长先减小后增大,在3 h时分别达到最致密的网络结构（P＜0.05）。对照组和添加MTGase的鲢鱼糜凝胶分别在40 ℃凝胶化3～6 h或2～4 h时凝胶特性较好,通过凝胶化时间的调节可控制鱼糜凝胶的结构演化方向,获得高品质鱼糜制品。     

Effects of Bambara Groundnut Protein Isolates and Microbial Transglutaminase on Textural and Sensorial Properties of Surmi Gel from Sardine (Sardinella albella)

Effects of different bambara groundnut protein isolates (BGPIs) at a level of 6 % (w/w) in combination with microbial transglutaminase (MTGase) at a concentration of 0.6 U g^?1 surimi on gel properties of sardine (Sardinella albella) surimi were investigated. In the absence of MTGase, all BGPIs showed the adverse effect on gel-forming properties of surimi, as evidenced by the decreases in breaking force and deformation (P?<?0.05). When MTGase was incorporated, the increases in breaking force and deformation were found for all BGPIs used. Water-holding capacity of all gels was improved when BGPIs were added in combination with MTGase (P?<?0.05). Whiteness of gels slightly decreased with the addition of BGPIs; however, MTGase had no impact on whiteness (P?>?0.05). Surimi gel added with BGPI prepared from defatted flour with heat treatment in the presence of ethylenediaminetetraacetic acid (DF-BGPI-HE) and MTGase showed well-ordered network and exhibited the lowest peroxide value and thiobarbituric acid-reactive substances than those containing other BGPIs. Gel containing DF-BGPI-HE had negligible beany flavour. Additionally, DF-BGPI-HE had the lower amount of volatile compounds after storage of 30 days at room temperature than other BGPIs. Thus, the addition of DF-BGPI-HE and MTGase was an effective means to render sardine surimi gel with improved gel property and caused no beany flavour in resulting gel.     

Gel‐forming ability of surimi from grass carp (Ctenopharyngodon idellus): influence of heat treatment and soy protein isolate

The effects of setting conditions and soy protein isolate (SPI) on textural properties of surimi produced from grass carp were investigated. Effects of setting temperature, setting time and protein concentration on the breaking force and distance were evaluated and compared utilizing response surface methodology. Models for breaking force and breaking distance of grass carp surimi were established. Protein concentration was the major factor affecting the gel strength of grass carp surimi. Breaking force and distance of grass carp surimi gels decreased with increase of protein ratio from SPI at 30 °C and 40 °C for 60 min setting and heating at 85 °C for 30 min, but the breaking force obtained for addition of 100 g kg^?1 SPI protein to grass carp surimi was higher than that for surimi alone at 60 °C for 60 min incubation and heating at 85 °C for 30 min. Copyright © 2005 Society of Chemical Industry     

Effect of formaldehyde on protein cross-linking and gel forming ability of surimi from lizardfish induced by microbial transglutaminase

Impact of formaldehyde (FA) at various levels (0–9 μmol/g surimi) on gel properties of surimi from lizardfish added with microbial transglutaminase (MTGase) was studied. During iced storage of 10 days, total and free FA in lizardfish flesh increased continuously (P < 0.05). In the presence of FA, breaking force of gels slightly increased, whilst the deformation decreased (P < 0.05). The addition of MTGase (0.4 units/g surimi) was able to increase gel strength and water holding capacity of resulting gel. Nevertheless, gel strengthening effect of MTGase was lowered when FA at higher level was present. Myosin heavy chain (MHC) dominantly underwent polymerisation to a higher extent when either MTGase or FA was added. The higher reduction in ε-amino group content was observed in natural actomyosin (NAM) when FA at higher levels (0–30 μmol/g protein) was incorporated. Acyl transfer reaction mediated by MTGase was impeded in NAM containing FA, especially at higher levels. Generally, FA had an adverse effect on cross-linking ability towards surimi proteins induced by MTGase. Therefore, cross-linking and gel-forming ability of lizardfish surimi could be maximised by MTGase when surimi contained no FA.     

ɛ-(γ-Glutamyl)lysine Crosslink Formation in Sardine Myofibril Sol during Setting at 25°C

The quantitative change of ?-(y-glutamyl)lysine (EGL) crosslink and relationship between crosslink content and gel-strength were examined on salt-ground myofibril sol from sardine (Sardinops melanostictus) during incubation at 25°C. In the presence of EGTA, no EGL crosslinks were detected in myofibril sol and gelation did not occur. The EGL crosslink content and breaking strength of gels increased in proportion to incubation time. High correlation was observed between the logarithm of breaking strength and logarithm of EGL crosslink content (r=0.987). The EGL crosslinks formed by transglutaminase are important in the setting of sardine meat sol at < 30°C.     

Gel Forming Characteristics of Frozen Surimi from Chum Salmon in the Presence of Protease Inhibitors

When salted surimi paste of chum salmon was incubated at 20–60°C, a marked loss of the breaking strength of heat-induced gel occurred simultaneously with breakdown of myosin heavy chain, but this was effectively suppressed by addition of cysteine protease inhibitors or bovine plasma powder. In the presence of protease inhibitor, the surimi gels were formed at relatively low temperatures showing highest gel strength at incubations of 50 and 60°C. Chum salmon surimi showed no evidence of suwari and no myosin heavy chain cross-linking.     

鸡蛋清蛋白对微生物转谷氨酰胺酶诱导鲢鱼鱼糜凝胶形成的影响

通过对鲢鱼鱼糜凝胶特性和溶解率的测定及SDS-PAGE检测研究鸡蛋清蛋白改善鱼糜凝胶特性的机理及其对微生物转谷氨酰胺酶(MTGase)诱导鱼糜凝胶形成的影响。结果表明：鸡蛋清蛋白(EA)和MTGase均能显著提高鱼糜凝胶特性。EA会降低MTGase诱导鱼糜凝胶的形成,显著降低MTGase诱导鱼糜凝胶的凝胶强度和破断强度,但不阻碍MTGase对肌球蛋白重链(MHC)的交联。     

Microbial Transglutaminase Affects Gel Properties of Golden Threadfin-bream and Pollack Surimi

ABSTRACT: The properties of surimi gels from threadfin-bream and pollack surimi set at 30 °C or 45 °C with microbial transglutaminase (MTGase) from Streptoverticillium ladakanum were determined. The optimal amounts of MTGase and setting conditions were: 0.3 unit/g surimi either at 30 °C for 90 min or at 45 °C for 20 min for threadfin-bream, and 0.2 unit/g surimi at 30 °C for 60 min for pollack. The strength of golden threadfin-bream surimi gels with 0.35 unit MTGase set at 30 °C for 90 min or 45 °C for 20 min was 3400 g cm, almost 3-fold of the control. SDS-PAGE analyses indicated that inter- and/or intramolecular cross-linking formed in the myosin heavy chain of MTGase-containing surimi gels.     

Improvement of Gelling Properties of Lizardfish Mince as Influenced by Microbial Transglutaminase and Fish Freshness

ABSTRACT:  The effects of microbial transglutaminase (MTGase) at different levels (0 to 0.8 units/g sample) on the properties of gels from lizardfish ( Saurida undosquamis ) mince set at 25 °C for 2 h or 40 °C for 30 min prior to heating at 90 °C for 20 min were studied. Breaking force and deformation of gels increased with increasing MTGase amount added ( P < 0.05). At the same MTGase level used, gels with the prior setting at 40 °C for 30 min showed a higher breaking force compared with those subjected to prior setting at 25 °C for 2 h ( P < 0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoretic study revealed that myosin heavy chain (MHC) underwent polymerization to a higher extent in the presence of MTGase. Regardless of setting condition, microstructure of gel added with MTGase was finer with a smaller void compared with that of gel without MTGase. Therefore, setting temperature affected the property of gels added with MTGase. Gel properties of mince obtained from lizardfish stored in ice for different times (0 to 10 d) with and without MTGase at a level 0.6 units/g were determined. Irrespective of MTGase addition, breaking force and deformation of all gels decreased as the storage time of lizardfish increased ( P < 0.05). The addition of MTGase was able to increase both breaking force and deformation of the resulting gel produced from lizardfish kept in ice for all storage times used. Therefore, both freshness and MTGase addition had the direct impact on gel properties of lizardfish mince.     

Effect of microbial transglutaminase on autolysis and gelation of lizardfish surimi

In the absence of microbial transglutaminase (MTGase), the textural properties of lizardfish surimi (Saurida spp) improved when pre‐incubated at 4 and 25 °C for 24 and 4 h, respectively. MTGase optimally catalyzed incorporation of monodansylcadaverine (MDC) into surimi at 40 °C. Addition of MTGase appeared to reduce autolytic activity at 25 and 40 °C, but had no effect on autolytic activity at 65 °C. Breaking force and deformation of lizardfish surimi significantly improved when 0.1 unit MTGase g^?1 surimi (1.8 g kg^?1) was added and pre‐incubated at either 25 or 40 °C. Textural properties improved concomitant with cross‐linked polymers of myosin heavy chain and tropomyosin, but not actin. Addition of MTGase also improved the storage modulus (G′). The gel network of surimi mixed with MTGase and pre‐incubated at 40 °C readily formed during the pre‐incubation period, while formation of the gel network began at 48.1 °C in the absence of MTGase. Copyright © 2005 Society of Chemical Industry     

Original article: Gel properties of red tilapia surimi: effects of setting condition,fish freshness and frozen storage

This study aimed to determine effects of setting condition, fish freshness and storage time of frozen surimi on properties of red tilapia surimi gel. To investigate the effect of setting condition, a combination of eight setting temperatures (35–70 °C) and four setting times (30–120 min) was used. Maximum breaking force, deformation and gel strength were obtained after the gel had been set at 40 °C for 90 or 120 min. Setting at 65 °C resulted in the lowest obtained gel strength, because of proteolytic degradation of myosin heavy chain. Increasing storage time of raw fish material in ice caused a significant decrease in gel strength of the resultant surimi gel (P < 0.05). Gels produced from surimi kept in frozen storage for up to 9 months also exhibited reduced gel strength, with a concomitant increase in the expressible drip, with increasing storage time (P < 0.05).     

Effect of soy protein isolate on gel properties of Alaska pollock and common carp surimi at different setting conditions

The effects of soy protein isolate (SPI) on the gel properties of different grade Alaska pollock and common carp surimi at different setting conditions were evaluated and compared. Breaking force and distance of gels decreased with increasing SPI concentrations in direct cook (85 °C for 30 min) and in cook after setting at 30 °C for 60 min conditions. The effect of SPI on gel strength of common carp surimi was less than in Alaska pollock surimi. The breaking force obtained for addition of 10% SPI to Alaska pollock surimi was higher than for surimi alone when cooked after incubation at 50 °C for 60 min. Addition of SPI decreased the whiteness and increased the yellowness of the gel. The gel structure showed that the addition of SPI modified the microstructure of the fish protein gel, thus resulting in surimi with different gelling properties. Copyright © 2004 Society of Chemical Industry