Enzymatic modification of selected functional properties of myofibril preparation obtained from mechanically recovered poultry meat

The effect of addition of 3 g/L of commercially available transglutaminase preparation to protein extracts obtained from mechanically recovered poultry meat was studied. The content of free thiol groups (–SH), thermal drip and gel texture were determined. After pre-incubation at 7–8 °C for 1, 3, 5 and 24 h, the samples were subjected to one-step heating at 50, 60, 70 and 80 °C and two-step heating at 50/80, 55/80 and 60/80 °C. The addition of preparation and the extension of pre-incubation time led to decrease of free -SH groups content. After heating, the number of thiol groups decreased, the texture was improved, but thermal drip from gels increased. The amount of –SH groups in gel extracts subjected to one-step heating decreased with simultaneous increase of mechanical strength of gels. Protein gels subjected to two-step heating exhibited higher firmness than gels subjected to one-step heating. Thus, the 3 g/L addition of transglutaminase preparation in combination with one-step thermal processing at 70 °C and pre-incubation for 3 h contributed to improvement of texture properties of model gels and low thermal drip.     

Analysis of texture and dynamics of water binding in enzymatically modified myofibrillar preparation obtained from washed mechanically recovered poultry meat

Experiments were conducted on a myofibrillar preparation, obtained from washed mechanically recovered poultry meat. An enzymatic preparation containing microbial transglutaminase (MTG) was added to samples of the myofibril isolate. The binding of water contained in the protein preparation with added MTG was assessed using low field nuclear magnetic resonance (NMR). Moreover, texture was analyzed in myofibril samples with the addition of transglutaminase pre-incubated for 0.5, 1.5, 3, 4.5 and 6 h. All measurements were taken at 7 ± 0.2 °C. Samples with added transglutaminase exhibited improved mechanical failure strength and better water binding capacity. The most dynamic increase of texture parameter values was observed in the interval from 1.5 to 3 h pre-incubation of the preparation with the added enzyme. Based on NMR (T ₁) testing it was established that the highest amount of water was bound by protein in the period from approximately 1 to 1.5 h pre-incubation. After that time free water content in the sample was again found to increase. This means that water was displaced from the system as a result of protein–protein interactions dominating over protein–water interactions. The above suggests that the enzymatic modification of the protein preparation contributed to the intensification of cross-linking between proteins in the preparation.     

焦磷酸钠对氧化猪肉肌原纤维蛋白的谷氨酰胺转移酶交联反应及凝胶性能的影响

以猪肉肌原纤维蛋白为研究对象,考察焦磷酸钠(sodium pyrophosphate,TSPP)对微生物谷氨酰胺转移酶(microbial transglutaminase,MTG)催化天然和氧化肌原纤维蛋白交联的程度及后续蛋白凝胶特性的影响。结果表明,肌原纤维蛋白的氧化程度随着H2O2溶液浓度的升高而增加,但TSPP在一定程度上可抑制羟自由基对蛋白的攻击,且TSPP处理(+TSPP)样品的MTG交联程度明显高于未经TSPP处理(-TSPP)样品。无论是否添加TSPP,随着H2O2溶液浓度的升高,蛋白凝胶的储能模量(G’)和凝胶强度均逐渐降低,加入MTG后均有所回升,且增幅随着H2O2溶液浓度升高逐渐降低。相对于未经TSPP处理样品,TSPP处理条件下未氧化样品的G’和凝胶强度增幅更高,但氧化时增幅降低速度更快。说明虽然TSPP处理有利于MTG交联引起的凝胶性能提高,但当受到氧化胁迫时,TSPP处理反而对其不利。     

Differentiation in improvements of gel strength in chicken and beef sausages induced by transglutaminase

This research investigated the improvement in the texture of chicken and beef sausages induced by using microbial transglutaminase (MTG). The ε-(γ-glutamyl)lysine (G-L) content and the extractability of myofibrillar proteins from these sausages were also investigated. Treatment with MTG significantly affected the breaking strength score in both meat types, especially for beef cooked at 80°C (p<0.001). The protein concentration of both meat types treated with MTG and extracted in water-soluble protein solution (WSP) was slightly decreased; compared with a significant decrease (p<0.003) in samples extracted in Guba-Straub-ATP solution (GS-ATP). The variation in protein extractability of both meat types could lead to some considerations of the mechanisms and the high affinity reaction between MTG and myosin heavy chain (MHC). SDS-PAGE analysis revealed significant changes in the density of the bands after adding MTG, especially for the beef samples. The G-L content in the presence of MTG was double that in control samples of both meat types. The amount of crosslinking in chicken and beef meat was different and found to be reasonable. Collectively, this suggests that the binding ability of myofibrillar proteins with MTG is strong and dominated by MHC. There was a unique reaction among MHC proteins with MTG molecules considered as a very advantageous reaction. This leads us to suggest that the functional properties of MTG make it a beneficial protein-binding agent, positively helping the functionality of proteins to improve the texture and gelation of meat products that are treated mechanically, such as sausages. Some variation in gel improvement level between chicken and beef sausages was observed; this resulted from the variation in meat proteins in response to MTG, as well as to the original glutamyl and lysine contents.     

Effect of non-meat proteins on hydration and textural properties of pork meat gels enhanced with microbial transglutaminase

The combined effect of incorporation of four non-muscle proteins, NMP (blood plasma, BP; sodium caseinate, SC; soy protein isolate, SPI; gelatin, G) at 2 g/100 g levels on hydration and textural characteristics of pork gels processed without or with 0.6 g/100 g microbial transglutaminase preparation (MTG) was investigated. Addition of SC and BP most favourably affected hydration properties and thermal stability, yielding lower cooking loss and expressible moisture for pork gels. Interactions between NMP and MTG were observed. Improvement of gel strength by addition of transglutaminase was observed for treatments containing SC and BP but not G nor soy isolate. Of the four proteins tested SC was found to be a superior substrate for MTG in enhancing textural properties of a gelled meat system. None of the tested ingredients was able to yield gel cohesiveness equivalent to the control containing 8% muscle proteins. Results of this study indicate a potential for using MTG to improve or modify the functional and textural properties of investigated food proteins (SC and BP in particular) in comminuted meat products.     

Effect of microbial transglutaminase on NMR relaxometry and microstructure of pork myofibrillar protein gel

The effect of microbial transglutaminase (MTG) on nuclear magnetic resonance relaxation (NMR) behaviour, water holding capacity (WHC) and microstructure of pork myofibrillar protein (PMP) gel was studied. The enzymatic protein preparation had significantly lower values of spin–spin relaxation time (T ₂), higher WHC and more porous microstructure in comparison with the control system. T ₂ was reduced from 226 ms (peak value) of the PMP gel containing no MTG to 188 ms of the PMP gel containing 2 U/g protein. However, no further decrease was shown when the concentration of MTG increased. The reduction was attributed to reduced mobility of water protons in the system. Scanning electron micrographs (SEM) showed the mobility of water in the proteins gel network was related to gel microstructure.     

Raman spectroscopic study of the effects of microbial transglutaminase on heat-induced gelation of pork myofibrillar proteins and its relationship with textural characteristics

The effects of microbial transglutaminase (MTG) on heat-induced gelation of pork myofibrillar proteins (PMP) structural changes, textural properties were studied by Raman spectroscopy and texture profile analysis (TPA), respectively. And the relationships between the structural changes and textural characteristics were estimated by principal component analysis (PCA). Changes in the Raman spectra were interpreted as the occurrence of secondary structural changes in myofibrillar proteins with MTG added. Modifications in the amide I (1600-1700 cm^− 1) regions indicated a significant (p < 0.05) decrease in ??-helix content, accompanied by a significant (p < 0.05) increase in ??-sheets, ??-turns and random coil content due to the addition of the enzyme. Obvious texture property changes were also determined by TPA. All these changes showed a strong, irreversible heat-induced gel formed due to the addition of MTG. The application of a dimensionality reducing technique such as PCA proved to be useful to determine the most influential properties of heat-induced gel. Significant (p < 0.05) correlations were found between these structural changes and the textural characteristics (hardness) in PMP system with the addition of MTG by PCA. The hardness was related positively to fraction of ??-sheet, ??-turns and random coil, and negatively to normalized intensity of 760 cm^− 1 and fraction of ??-helix. The samples are closely grouped in a cluster defined by level of MTG.     

Evaluation of red bean protein [Vigna angularis] isolate on rheological properties of pork myofibrillar protein gels induced by microbial transglutaminase

The effects of 1% red bean protein isolate (RBPI) on the gel properties of myofibrillar protein (MP) in various levels of microbial transglutaminase (MTG: 0%, 0.1%, 0.5%, & 1%) were evaluated. The cooking yield (CY) of the MP gels decreased with increasing MTG level, while the addition of RBPI improved the CY of the MP gels. Gel strength (GS) was also improved when RBPI was incorporated into the MP gels containing higher than 0.5% of MTG. The addition of MTG and RBPI was slightly changed the endothermic peak temperatures. Scanning electron microscopy (SEM) showed that the three‐dimensional structure of MP with RBPI alone or in combined with MTG was compacted as compared to the control. Based on the results, RBPI could be functioned as a substrate for MTG (0.5–1.0%) and a water binder of meat protein gel mediated by MTG.     

Gelation properties of myofibrillar/pea protein mixtures induced by transglutaminase crosslinking

Gelation properties of mixtures of myofibrillar protein isolate (MPI)/pea protein isolate (PPI) were studied using a dynamic oscillatory rheometer and a texture analyzer to evaluate PPI as a possible meat product additive. The inclusion of microbial transglutaminase (MTG) increased the gel strength of MPI/PPI mixture (3% + 1%) more than it did for MPI (3%), but less than a 3% MPI, 1% soy protein isolate combination. The direct evidence of interaction between muscle and pea proteins in the form of new sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) bands was not found; however, the improvement in gel strength or gel peak force for the MPI/PPI mixture (3% + 1%) with inclusion of MTG suggested that some ? (γ-glutamyl) lysine (G-L) crosslinking occurred between muscle and pea proteins. It likely that pea protein acted as a non-gelling component and interspersed throughout the primary MPI gel network and the addition of MTG promoted partial crosslinking of MPI. Consequently, MTG is useful in improving gelation properties of heat-induced MPI/PPI gel.     

Gelation properties of chicken myofibrillar protein induced by transglutaminase crosslinking

Gelation properties of chicken myofibrillar protein isolate (MPI) and the effect of microbial transglutaminase (MTG) were studied using a dynamic oscillatory rheometer and a texture analyzer. Final heating temperature had a great impact on gel stiffness and the maximum gel stiffness was obtained at 95 °C. pH and ionic strength also influenced gel stiffness and the maximum gel stiffness was achieved at pH 6, 0.9 M NaCl; however, less stiff gels were formed in 0.6 and 1.2 M NaCl. In the MPI concentration range of ∼0.5-5%, a positive correlation was observed between gel stiffness or gel peak force and MPI concentration. When MTG was included at levels of ∼0 to 12-15 U, positive linear relations were found between gel stiffness or peak force and MTG levels. However, negative correlations for these parameters were observed at higher MTG concentrations. When MTG level was greater than 15 U, gel stiffness or peak force tended to decrease. The improvement in gel strength or gel peak force for the MPI with inclusion of MTG suggested that some ε (γ-glutamyl) lysine (G-L) crosslinking occurred among myofibrillar molecules. Thus, MTG is useful in improving gelation properties of heat-induced MPI gel and provides new opportunities to expand the utilization of low value meat in muscle foods.     

Effect of selected commercial substances with cryoprotective activity on the quality of mechanically recovered,washed and frozen stored poultry meat

Investigations were conducted on mechanically recovered poultry meat (MRPM) and on protein preparation obtained from MRPM by washing it first with 1% water solution of sodium chloride and with water afterwards. The raw materials were frozen at the temperature of –23°C. The effect of added stabilizers on the quality of gels produced from fresh raw materials, and after freezing and frozen storage was assessed. The following additives were used: 1% pork hydrolizate (Pork Stock( 0.5% Cremodan containing carrageens, and 1.5% bovine blood plasma (AMP 600N). Freezing and frozen storage caused a significant reduction of functional properties of MRPM and its protein preparation. None of the examined additives protected simultaneously all the investigated functional properties of the frozen samples. The amount of thermal drip, the gel texture and the amount of protein transition heat were determined by scanning differential calorimetry. The lowest thermal drip in gels obtained from frozen‐stored samples was observed when bovine blood plasma was used as a stabilizer. On the other hand, the most advantageous protective effect on the proteins of the frozen MRPM and on the preparation, determined by mechanical strain resistance of the gels, was found with 1% pork hydrolizate added. The results of thermodynamic investigations of proteins revealed that the best protective effect on the frozen preparation was observed with 1.5% blood plasma added. No protective activity of added Cremodan on proteins of the frozen protein preparation was noted.     

Effect of microbial lipase and transglutaminase on the textural,physicochemical, and microbial parameters of fresh quark cheese

In this study, the addition of microbial transglutaminase (MTG) and lipase in quark cheese samples was studied during storage (21 d). Four types of cheese were made using 3 different levels of MTG (T1, 0.1 g/L; T2, 0.2 g/L; T3, 0.3 g/L) and lipase (T1, 0.02 g/L; T2, 0.04 g/L; T3, 0.06 g/L), and one cheese was made without any treatment as a control sample. The physicochemical, textural, microbial, and sensory properties of cheese samples were monitored at 1, 7, 14, and 21 d of storage period. The results showed that the treated samples had higher proteolysis and lipolysis activities during storage than the control sample. The textural analysis indicated an insignificant increase in the hardness value of the enzyme-treated sample. Also, the sensory analysis exhibited that the treated samples had higher texture acceptability. The higher concentration of enzymes resulted in lower color, odor, taste, and overall acceptability, and higher microbial population. Finally, the addition of microbial MTG and lipase in preparation of quark cheese samples could be recommended for a short storage time.     

Effect of selected commercial substances with cryoprotective activity on the quality of mechanically recovered,washed and frozen stored poultry meat

Investigations were conducted on mechanically recovered poultry meat (MRPM) and on protein preparation obtained from MRPM by washing it first with 1% water solution of sodium chloride and with water afterwards. The raw materials were frozen at the temperature of -23 C. The effect of added stabilizers on the quality of gels produced from fresh raw materials, and after freezing and frozen storage was assessed. The following additives were used: 1% pork hydrolizate (Pork Stock), 0.5% Cremodan containing carrageens, and 1.5% bovine blood plasma (AMP 600N). Freezing and frozen storage caused a significant reduction of functional properties of MRPM and its protein preparation. None of the examined additives protected simultaneously all the investigated functional properties of the frozen samples. The amount of thermal drip, the gel texture and the amount of protein transition heat were determined by scanning differential calorimetry. The lowest thermal drip in gels obtained from frozen-stored samples was observed when bovine blood plasma was used as a stabilizer. On the other hand, the most advantageous protective effect on the proteins of the frozen MRPM and on the preparation, determined by mechanical strain resistance of the gels, was found with 1% pork hydrolizate added. The results of thermodynamic investigations of proteins revealed that the best protective effect on the frozen preparation was observed with 1.5% blood plasma added. No protective activity of added Cremodan on proteins of the frozen protein preparation was noted.     

添加牛血浆蛋白和MTG酶对乳化肠凝胶特性的影响

研究了牛血浆蛋白（BPP）和谷氨酰胺转胺酶（MTG）添加量对乳化肠产率、凝胶破裂强度和硬度、色泽的影响。结果表明,添加牛血浆蛋白和MTG酶对乳化肠产率、总压出汁液、凝胶强度和硬度有显著影响,但牛血浆蛋白处理组与MTG酶处理组有一定差异。添加牛血浆蛋白处理组乳化肠的L＊、a＊和b＊值显著高于MTG处理组。结论指出,牛血浆蛋白作为一种天然蛋白添加剂用来代替MTG,不仅能降低加工成本,还可以改善肉制品的品质。     

磷酸盐对PSE 猪肉肌原纤维蛋白溶胶及凝胶性质的影响

PSE 肉的肌原纤维蛋白质(MP)的溶解度比正常肉(red, firm and non-exudative;RFN)低、凝胶性质比RFN差,本研究探讨磷酸盐对PSE 猪肉凝胶特性的影响。以猪背最长肌PSE 和RFN 肉为材料,采用分光光度法、质构测定法等研究六偏磷酸钠(SHMP)、焦磷酸钠(SPP)、三聚磷酸钠(TPP)对MP 溶解特性、聚集性以及凝胶强度的影响。结果表明,磷酸盐对PSE 和RFN 提取MP 的溶解度、蛋白质聚集性和凝胶强度均有显著的影响,TPP 添加量为0.25% 时PSE 的凝胶强度接近未添加TPP 的RFN 的凝胶强度。磷酸盐可改善体系环境,改善PSE 蛋白质溶胶及凝胶的功能特性。     

Effect of salt addition on the thermal behavior of proteins of bovine meat from Argentina

Research was undertaken to investigate how the addition of sodium chloride (NaCl) and/or sodium tripolyphosphate (TPP) to sous vide cooked meat pieces produces an increase in water holding capacity (WHC). Semitendinosus muscles were injected to obtain tissue final concentrations of 0.70% NaCl, 0.25% TPP, 0.70% NaCl+0.25% TPP, and 1.20% NaCl+0.25% TPP. SDS-PAGE analysis showed increased protein solubilization in those treatments which included NaCl. Thermal analysis of whole muscles and isolated myofibrils showed the destabilizing effect of NaCl and a global stabilizing effect of TPP. Both salts together induced a destabilizing global effect, where TPP assisted NaCl in breaking the meat structure. It is suggested that the WHC increments are related to conformational changes in myofibrillar proteins and to the weakening of myofibrillar structure by the removal of myofibrillar proteins.     

Enzymatic modification of protein preparation obtained from water-washed mechanically recovered poultry meat

Studies were conducted on a protein preparation obtained from washed mechanically recovered poultry meat (MRPM). The effect of addition of 3 g/kg microbial transglutaminase (MTG) to poultry meat protein was evaluated in terms of texture changes by dynamic mechanical analysis (DMA) and nuclear magnetic resonance (NMR) to determine water content in the preparation and its effect on protein. Samples with the addition of MTG were pre-incubated at 5–6 °C for 1.5, 3, 4.5, 6, 7.5, 9 and 24 h. The largest changes for both texture parameters and rheological properties were observed in the interval of approx. 4–7 h incubation. The protein preparation with the enzyme added had significantly higher values of the moduli of elasticity (G₁) and losses (G₂) in comparison to the control system. Samples with the addition of MTG also showed a higher water-binding capacity. From the NMR studies it was found that the greatest amount of water was bound by protein in the period of approx. 2.5–5 h incubation. After that time an increase was found in the amount of free water in the sample, which suggests that it was displaced from the system by stronger protein–protein bonds.     

Dependence of microbial transglutaminase on meat type in myofibrillar proteins cross-linking

The objectives of this study were to determine the factors that cause differences in the improvements of gel strength and ε(γ-glutamyl)lysine (G-L) content in chicken and beef (Japanese black cattle) myofibrillar proteins after adding microbial transglutaminase (MTG). As the amount of MTG added increased, the breaking strength increased progressively (p < 0.01) in chicken and beef samples, with the exception of chicken samples treated at 40 °C. The values of elasticity in the chicken samples were lower than those of the beef samples (p < 0.01). Surprisingly, the elasticity level, ε(γ-glutamyl)lysine contents and myosin heavy chain (MHC) band sizes of chicken and beef at all levels of MTG were significantly different (p < 0.01). The results of this study suggest that MTG activity was affected by MTG inhibitors; that MTG develops the texture of myofibrils differently in different species. However, the activity is limited and inconstant among meat proteins, as suggested by the data collected from the chicken samples. As a result, when the transferable amino acid residues are depleted (cross-linked) by MTG activity, the function of MTG will be insignificant. The correlation between MTG and different sources of meat protein is quite unstable but it is strong, which was observed when chicken and beef responded differently to MTG because their chemical and physiological properties were different. The remarkable rate of formation of cross-linked proteins and the discrepancy between the expected and observed amount of dipeptide raises the possibility that there are enzymes capable of reversing the reaction induced by transglutaminase in chicken and beef myofibrils. In summary, our results suggest that access of MTG to chicken and beef myofibrils is different because it depends on physiological (muscles and their fibre types), biological (substrates) and biochemical (inhibitors and amino acids) variables.     

聚葡萄糖对大米淀粉凝胶老化特性的影响

为探究聚葡萄糖(PD)对大米淀粉(RS)凝胶老化特性的影响,利用快速黏度分析仪(RVA)、差示扫描量热分析(DSC)、X-射线衍射分析(XRD)和扫描电子显微镜(SEM)研究聚葡萄糖对大米淀粉糊化黏度特性、热特性、晶体结构和微观结构的影响。结果表明,随着聚葡萄糖添加量的增大,大米淀粉糊化的峰值黏度、崩解值和回生值均显著下降(p<0.05);但是糊化温度升高,大米淀粉的糊化焓值和老化焓值均显著降低(p<0.05),大米淀粉凝胶的相对结晶度从14.09%降到8.61%。SEM结果表明添加聚葡萄糖后,大米淀粉凝胶孔洞变小,表面更致密、光滑,说明聚葡萄糖可以抑制大米淀粉凝胶的重结晶,延缓大米淀粉凝胶的老化。     

不同质量比鸡胸肉、鸡腿肉混合肌原纤维蛋白的热诱导凝胶特性

以鸡胸肉和鸡腿肉为材料,研究鸡胸、鸡腿肉混合肌原纤维蛋白不同质量比(9:1、3:1、1:1、1:3、1:9)热变性温度、质构特性、流变特性和保水性的差异。结果表明：鸡胸肉肌原纤维蛋白的凝胶特性显著优于鸡腿肉,鸡胸肉与鸡腿肉的混合可以显著提升鸡腿肉蛋白的硬度、保水性;混合蛋白中,m鸡胸肉蛋白:m鸡腿肉蛋白为9:1的蛋白硬度、保水性最大,分别达到167.64g和83.6%,其凝胶特性和纯鸡胸肉差异不大;m鸡胸肉蛋白:m鸡腿肉蛋白为1:9的混合蛋白硬度、保水性和贮能模量终值均为最小,分别为110.82g、48.75%和297Pa,其凝胶特性优于纯鸡腿肉肌原纤维蛋白。