Gel‐enhancing effect and protein cross‐ linking ability of tilapia sarcoplasmic proteins

BACKGROUND: Tilapia (Oreochromis niloticus) sarcoplasmic proteins contain substantial transglutaminase (TGase) activity. The enzyme catalyzes the protein cross‐linking reaction, resulting in a more elastic gel. The objective was to investigate the gel‐enhancing effect of sarcoplasmic proteins from tilapia as related to TGase activity. RESULTS: Total TGase activity of sarcoplasmic proteins concentrate (SpC) increased about 3.6‐fold after ultrafiltration using 30 kDa membrane, but specific activity remained unchanged, indicating minimal TGase purification by ultrafiltration. Addition of 1 mg mL^?1 SpC containing 40 units TGase activity induced cross‐linking of tilapia actomyosin, and the extent of cross‐linking increased with added level of SpC. Myosin heavy chain (MHC) and troponin were preferably cross‐linked by tilapia SpC, while actin and tropomyosin were not affected. Higher retention of MHC was observed concomitantly with greater content of cross‐linked protein when SpC was added to lizardfish surimi. Lizardfish surimi with 10 g kg^?1 SpC added and pre‐incubated at 37 °C for 1 h exhibited 91.6% and 26.7% increase in breaking force and deformation, respectively, when compared to the control. CONCLUSIONS: Residual TGase activity in SpC played an important role in catalyzing the protein cross‐linking and enhancing actomyosin gelation. SpC could be a potential ingredient for improving textural properties of fish protein gel. Copyright © 2007 Society of Chemical Industry     

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.     

The effect of transglutaminase on some physicochemical and sensory properties of the Turkish drinking yoghurt Ayran

This article presents research on the effect of microbial transglutaminase on the properties of Turkish drinking yoghurt Ayran. Transglutaminase (TGase) enzyme was added to Ayran milk at different production steps, after pasteurisation and together with starter culture addition, and two different incubation times (10 min and 1 h) were used. Three TGase‐treated Ayran samples and control sample were analysed on 1st, 10th and 20th days of storage. TGase addition did not cause significant changes on chemical properties of Ayran (P > 0.05). However, physical properties of Ayran were improved by TGase throughout the storage period. Compared to the untreated control sample, pretreatment of milk with enzyme increased the viscosity dramatically (P < 0.05) and prevented serum separation. Scanning electron microscopic studies revealed that transglutaminase treatment of milk led to more regular distribution of proteins in Ayran gel and decreased gel permeability. TGase treatment did not show any unfavourable effect on the sensory properties of the final product.     

Modification of pork-skin jelly by enzymatic cross-linking: melting resistance and quality improvement

Improvements of melting resistance and quality by modification of pork-skin jelly through enzymatic cross-linking were studied, and the mechanism of quality improvement was discussed in this work. Gel strength, springiness and chewiness of modified gel increased significantly (P < 0.05). Transglutaminase (TGase) also improved the viscoelasticity, stability and melting resistance of gel system, as proved by rheological analysis. Sensory evaluation showed that increase in texture need to be moderate and the utilisation of 0.6% TGase was the most appropriate for pork-skin jelly. Significant effects of TGase on inducing protein cross-link and aggregation were confirmed by determining rheology during enzyme treatment and cross-linking extent of pork-skin soup. Correlation analysis showed TGase could improve melting temperature and texture by facilitating cross-linking. Covalent interaction based on ε-(γ-glutamyl)-lysine induced by TGase could play the main role in these improvements. This study suggested that TGase could be applied to design gelatin-based food for tailored quality properties through enzymatic cross-linking.     

Effect of Transglutaminase on Physicochemical Properties of Set-style Yogurt

In this study, the effect of some ingredients such as skimmed milk powder, whey, sodium caseinate, calcium caseinate, whey protein concentrate (35, 60 kg/100 kg dry solids), whole milk powder, condensed milk and transglutaminase (TGase) on the properties of set-style yogurt was investigated. These protein and dry matter sources (2%) and TGase (1 U/g milk protein) were added into pasteurized milk and incubated prior to fermentation for 2 h at 40°C. After fermentation, enzyme action was stopped by heating for 1 min at 80°C. The control groups were conducted with addition of these materials into milk without TGase. All of the milk samples were inoculated with yogurt cultures at 45°C, until the pH was dropped to 4.4. Syneresis, gel-strength, acetaldehyde amounts, and the degree of TGase reaction were determined. As a result, yogurt products made from enzyme-treated milk showed increased gel strength and less syneresis. SDS-PAGE results showed that the enzyme TGase produced crosslink formation between different protein fractions of milk. In addition, it was also determined that TGase application caused a decrease in acetaldehyde amounts.     

Effect of using transglutaminase on physical,chemical and sensory properties of set-type yoghurt

This study was conducted in order to evaluate the effects of transglutaminase (TGase) addition on some properties of set-type yoghurts such as titratable acidity, lactic acid, tyrosine, viscosity, gel firmness, syneresis, aroma compounds, sensory analysis, and micro-structural properties. The enzyme was added to yoghurt-milk at different production steps (after homogenization, after pasteurization and together with starter culture addition) and two different incubation times (10 min and 1 h) were used. Five TGase treated yoghurt samples and control sample were analyzed on 1st, 10th, and 20th days of storage. TGase addition did not cause significant changes on chemical properties of yoghurts. However, enzyme addition after pasteurization increased the gel strength and decreased the syneresis. Results of electron microscope showed that enzyme addition led proteins to be distributed more evenly in gel network due to the formation of cross-links between proteins.     

Transglutaminase from Streptoverticillium ladakanum and application to minced fish product

The transglutaminase (TGase) from Streptoverticillium ladakanum was purified to electrophoretic homogeneity after ammonium sulfate fractionation and Blue Sepharose Fast Flow chromatography. The molecular weight of the purified TGase was 30.5 kDa estimated by Superdex 75HR gel filtration, and 37.5 kDa by SDS-PAGE. This enzyme, with optima at pH at 6.0 and 50°C was very stable at pH 5.0–7.0. It was strongly inhibited by PCMB, PMSF, Pb²⁺, Zn²⁺ and Cu²⁺, but not affected by EDTA and Ca²⁺. This suggested that the purified TGase was calcium-independent and its active center contained cysteine. It catalyzed the crosslinking of fish myosin heavy chain and substantially increased the gel strength of mackerel surimi.     

Improvement of gel strength and melting point of fish gelatin by addition of coenhancers using response surface methodology

Abstract: Fish gelatin is a potential alternative to mammalian gelatin. However, poor gel strength and low melting point limit its applications. The study was aimed at improving these properties by adding coenhancers in the range obtained from response surface methodology (RSM) by using Box–Behnken design. Three different coenhancers, MgSO₄, sucrose, and transglutaminase were used as the independent variables for improving the gel strength and melting point of gelatin extracted from Tiger‐toothed croaker (Otolithes ruber). Addition of coenhancers at different combinations resulted gel strength and melting point in the range of 150.5 to 240.5 g and 19.5 to 22.5 °C, respectively. The optimal concentrations of coenhancers for predicted maximum gel strength (242.8 g) obtained by RSM were 0.23 M MgSO₄, 12.60% sucrose (w/v), and 5.92 mg/g transglutaminase and for predicted maximum melting point (22.57 °C), the values were 0.24M MgSO₄, 10.44% sucrose (w/v), and 5.72 mg/g transglutaminase. By addition of coenhancers at these optimal concentrations in verification experiments, the gel strength and melting point were improved from 170 to 240.89 g and 20.3 to 22.7 °C, respectively. These experimental values agreed well with the predicted values demonstrating the fitness of the models. Results from the present study clearly revealed that the addition of coenhancers at a particular combination can improve the gel strength and melting point of fish gelatin to enhance its range of applications. Practical Application: There is a growing interest in the use of fish gelatin as an alternative to mammalian gelatin. However, poor gel strength and low melting point of fish gelatin have limited its commercial applications. The gel strength and melting point of fish gelatin can be increased by incorporation of coenhancers such as magnesium sulphate, sucrose, and transglutaminase. Results of this work help to produce the fish gelatin suitable for wide range of applications in the food industry.     

Emulsifying properties of the transglutaminase-treated crosslinked product between peanut protein and fish (Decapterus maruadsi) protein hydrolysates

BACKGROUND: Defatted peanut meal, a protein‐rich by‐product from the oil extraction industry, is underutilised owing to its inferior functional properties. In this study, transglutaminase (TGase) crosslinking and proteolysis were used to improve the emulsifying properties of peanut protein isolate (PPI) extracted from the meal. PPI and PPI hydrolysate (PPIH) were conjugated separately with fish (Decapterus maruadsi) protein hydrolysate (DPH), catalysed by TGase to obtain improvements in the emulsifying properties. RESULTS: Analyses by electrophoresis and high‐performance liquid chromatography indicated that polymers were formed in all TGase‐treated samples. In emulsions of PPIH, PPI‐DPH and PPIH‐DPH the volume/surface average particle diameter (d₃₂), creaming and instability phenomenon were decreased and the zeta‐potential was increased after TGase treatment, showing improved emulsifying activity and emulsion stability. In the case of PPI, TGase treatment had no effect on the emulsifying activity, but the emulsion stability of TGase‐treated PPI was improved. CONCLUSION: The study showed that TGase crosslinking and proteolysis could improve the emulsifying properties of PPI, while proteolysis followed by TGase crosslinking proved more efficient. The emulsifying properties of the heterologous protein systems of PPI‐DPH and PPIH‐DPH were also improved by TGase treatment. Copyright © 2010 Society of Chemical Industry     

谷氨酰胺转氨酶预交联改善CaSO4诱导大豆分离蛋白凝胶性的研究

酶法改性能够有效提升大豆蛋白的凝胶性。为了探讨谷氨酰胺转氨酶(transglutaminase, TGase)预交联对盐诱导大豆分离蛋白凝胶性的影响,通过控制酶浓度、预交联时间制备不同预交联程度的大豆分离蛋白(soy protein isolate,SPI)溶液,并研究其在CaSO₄作用下的成胶性能。结果显示,与未经TGase处理的SPI相比,TGase适度预交联能够显著提升SPI的凝胶品质。经3～5 U/g TGase预交联20 min或3 U/g TGase预交联20～30 min后,SPI凝胶性得到了不同程度的提升,其中弹性模量、屈服应力、屈服应变、持水率最大分别提高了124.5%、269.0%、135.0%及53.0%。然而,过度预交联产生过大的蛋白聚集体,导致最终形成的凝胶结构粗糙、多孔,凝胶强度、持水力等均显著下降(P<0.05)。由此可见,合理利用TGase对蛋白进行预交联处理能够改善SPI凝胶制品品质,对于TGase在食品工业中的应用及传统豆制品质构改良具有重要的指导意义。     

Improving the physical properties of fish gelatin by high hydrostatic pressure (HHP) and ultrasonication (US)

In this study, it was aimed to improve the physical properties of fish gelatin by using high hydrostatic pressure (HHP) and ultrasonication (US). Gelatin solutions were exposed to different pressures and ultrasonication separately and gelled afterwards. The physicochemical measurements based on gel strength, turbidity and rheology experiments showed that HHP treatment on fish and bovine gelatin stabilized the gelatin network by organising the structure and reducing the free volume. Both processing methods (HHP and US) increased the gel strength significantly (P < 0.05) compared with non-treated samples. Fourier-transform infrared spectroscopy (FTIR) results indicated that conformations of amino acids changed after the treatments. Furthermore, US treatment was shown to destroy the gelatin network, change the gelation mechanism and decreased the degree of aggregation. Both treatments improved the gel characteristics as gel strength, gelling and melting temperatures of the fish gelatin. At the end, the best combination for fish gelatin among HHP and US treatments was found as 400 MPa–10 °C–15 min pressurisation.     

Microencapsulation of Bifidobacterium bifidum F‐35 in Whey Protein‐Based Microcapsules by Transglutaminase‐Induced Gelation

Abstract: Bifidobacterium bifidum F‐35 was microencapsulated into whey protein microcapsules (WPMs) by a transglutaminase (TGase)‐induced method after optimization of gelation conditions. The performance of these WPMs was compared with that produced by a spray drying method (WPMs‐A). WPMs produced by the TGase‐induced gelation method (WPMs‐B) had larger and denser structures in morphological examinations. Native gel and SDS‐PAGE analyses showed that most of the polymerization observed in WPMs‐B was due to stable covalent crosslinks catalyzed by TGase. The degradation properties of these WPMs were investigated in simulated gastric juice (SGJ) with or without pepsin. In the presence of pepsin, WPMs‐A degraded more quickly than did WPMs‐B. Finally, survival rates of the microencapsulated cells in both WPMs were significantly better than that of free cells and varied with the microencapsulation method. However, WPMs‐B produced by TGase‐induced gelation could provide better protection for microencapsulated cells in low pH conditions and during 1 mo of storage at 4 °C or at ambient temperature.     

TG酶对高温杀菌鱼糜凝胶特性影响

高温杀菌处理会造成淡水鱼糜凝胶品质劣化。以白鲢鱼糜为原料,通过测定质构特性、凝胶强度、持水性、白度和流变特性,研究谷氨酰胺转氨酶(TG酶)对高温杀菌鱼糜凝胶特性的影响。结果表明,添加TG酶所制得鱼糜凝胶经121℃,10 min高温杀菌后,硬度、弹性、持水性、白度及流变特性均有较好的改善。与对照组相比,添加0.5%TG酶的鱼糜凝胶强度增加48.7%。红外光谱测定结果表明,TG酶的加入有利于蛋白质α-螺旋结构的维持,减少蛋白主要二级结构破坏,使鱼糜制品维持良好凝胶结构。     

High Pressure Effects on Gelation of Surimi and Turkey Breast Muscle Enhanced by Microbial Transglutaminase

High pressure effects on the strength (stress) and elasticity/deformability (strain) of surimi and turkey breast meat gels containing microbial transglutaminase (TGase) were evaluated. Pressurization of muscle proteins at 4°C prior to incubation at 25°C or 40°C (setting) increased gel strength 2–3 fold in uncooked surimi gels, but not in uncooked turkey gels. However, pressurization at 40°C or 50°C prior to setting increased the strength of turkey gels. Similar effects of prior pressurization, but of lesser magnitude, occurred in gels formed by directly or subsequently (following setting) cooking at 90°C. SDS-PAGE confirmed that myosin crosslinking occurred due to TGase activity during the setting treatment, which had survived prior pressure treatment. High pressure rendered protein substrates more accessible to TGase thereby enhancing intermolecular cross-link formation and gel strength.     

谷氨酰胺转移酶对明胶-CaCO3矿物质膜成膜性的影响

本实验通过向明胶-碳酸钙矿物质膜中添加谷氨酰胺转移酶（TGase）,研究了谷氨酰胺转移酶对明胶-碳酸钙矿物质膜特性的影响,对谷氨酰胺转移酶处理前后样品进行厚度、质构、水溶性、水蒸气透过系数、扫描电镜（SEM）、流变性、差示热量扫描（DSC）等方法表征。研究结果表明：在成膜溶液中加入谷氨酰胺转移酶（6 U/g）可以使矿物质膜的厚度增加19.69%、成膜液凝胶强度增加17.24%、膜的抗拉强度增加28.05%、断裂伸长率增加21.27%,而水溶性和水蒸气透过率没有显著改变;扫描电镜表明,谷氨酰胺转移酶交联的矿物质膜表面和断面与不加谷氨酰胺转移酶的矿物质膜相比更加粗糙;流变性结果表明,谷氨酰胺转移酶加入后成膜溶液的粘度显著增加;差示热量扫描表明,谷氨酰胺转移酶催化明胶-碳酸钙矿物膜产生了交联。     

Properties of Cassava Starch Modified by Amylomaltase from Corynebacterium glutamicum

Amylomaltase (α‐1,4‐glucanotransferase, AM; EC 2.4.1.25) from Corynebacterium glutamicum expressed in Escherichia coli was used to prepare the enzyme‐modified cassava starch for food application. About 5% to 15% (w/v) of cassava starch slurries were incubated with 1, 3, or 5 units of amylomaltase/g starch. Apparent amylose, amylopectin chain length distribution, thermal properties, freeze–thaw stability, thermo‐reversibility, and gel strength of the obtained modified starches were measured. The apparent amylose content and retrogradation enthalpy were lower, whereas the retrogradation temperatures, freeze–thaw stability, and thermo‐reversibility were higher than those of the native cassava starch. However, when amylomaltase content was increased to 20 units of amylomaltase/g starch and for 24 h, the modified starch showed an improvement in the thermo‐reversibility property. When used in panna cotta, the gel strength of the sample using the 20 units/24 h modified cassava starch was similar to that of using gelatin.     

Physicochemical properties of whole soybean curd prepared by microbial transglutaminase

Whole soybean curd (WSC) was manufactured using micronized full-fat soybean (MFS) powder and microbial transglutaminase (TGase). The WSC prepared with 15% MFS had typical soybean curd texture with a hardness of 513 dyne/cm². It was confirmed that 7S and 11S protein fractions as major soy proteins disappeared in SDS-PAGE. Also, WSC prepared with 15% MFS and 10% TGase had excellent textural properties with a hardness of 645 dyne/cm² and springiness of 0.98. Addition of 0.5% gelatin in WSC prepared with 15% MFS and 5% TGase resulted in higher hardness (708 dyne/cm²) and springiness (0.98), as well as the highest values of G′ and G″. The surface properties of WSC were observed using a SEM, indicating the interrelationship of higher hardness and compact protein network filled with small cells. It was concluded that WSC prepared after heat treatment of 15% MFS at 95°C for 5 min, followed by an enzyme reaction with 10% TGase for 1 h, had more enhanced hardness and springiness than commercial WSC. Despite the addition of 5% TGase, WSC with improved textural properties can be manufactured by the fortification of 0.5% gelatin.     

三种添加剂在鱼肉猪肉复合凝胶中的作用

为了改善鱼肉/猪肉复合肉糜（配比分别为3：7和7：3）的凝胶性能,研究了添加剂（TGase、CaCl2和焦磷酸钠）对鱼肉/猪肉复合凝胶的作用效果。结果表明,两种复合肉糜凝胶的破断强度、凝胶强度和保水性均随着TGase添加量的增加而增大,对鱼肉/猪肉（7：3）复合凝胶的增强作用更显著,而且降低了鱼肉/猪肉（7：3）复合凝胶的白度。CaCl2的添加也可增大复合肉糜的凝胶强度和白度,但高剂量的CaCl2（〉10mmol/kg）会降低其保水性。鱼肉/猪肉为7：3和3：7复合凝胶的凝胶强度分别于焦磷酸钠为0.05%和0.3%时达到最大值,高剂量的焦磷酸钠（〉0.3%）会提高复合凝胶的持水性,而对白度的影响较小。     

转谷氨酰胺酶对鳙鱼鱼糜凝胶特性的影响

研究了不同浓度的转谷氨酰胺酶 (简称TGase)对鳙鱼鱼糜凝胶特性的影响。结果表明 ,在鳙鱼鱼糜中添加不同浓度的TGase ,均可使其凝胶的破断强度、凹陷深度、凝胶强度及持水性增加 ,而对其颜色、白度无影响 ;酶的最佳浓度为 0 5 % ,在此浓度下 ,鱼糜的凝胶强度为 2 13 0 9g·cm ,是对照样的 4倍多 ;通过对SDS 聚丙烯酰胺凝胶电泳及溶解度的分析表明 ,TGase可催化鳙鱼鱼糜中的肌球蛋白重链 (MHC)形成共价交联键。电镜分析则表明 ,TGase的加入可使鳙鱼鱼糜形成致密、均匀的凝胶网络结构     

Gel-Forming Characteristics and Rheological Properties of Kefiran

A water soluble polysaccharide “kefiran” produced by Lactobacillus kefiranofaciens was examined for its gel-forming and rheological properties. Kefiran (3%) formed gel in the presence of ethanol (4–10%). The gel strength in 8% ethanol was comparable to that of 3% gelatin gel in water. Addition of casein (3%) increased gel strength 1.5–2.0 fold. The unique properties of kefiran may make it a useful food additive.