Effect of enzymatic cross-linking of milk proteins on functional properties of set-style yoghurt

This paper presents research on the effect of enzymatic cross-linking of milk proteins on the properties of yoghurt. Whole milk was incubated with transglutaminase (TG) prior to fermentation (2 h, 40°C, E/S ratio 1/2000). Enzyme action was stopped by heating (1 min, 80°C). Skim-milk was treated by simultaneous use of TG and thermophilic yoghurt starter culture without inactivation of the enzyme. A TG treatment of milk prior to fermentation led to prolonged fermentation, while the concomitant use of TG and culture had no influence on fermentation time. Post acidification of yoghurt during storage was lower for products from enzyme-treated milk. This applies both for products cross-linked prior to fermentation with enzyme inactivation, and for simultaneous use of culture and TG without inactivation of the enzyme. Scanning electron microscopic studies revealed that a TG treatment of milk led to reduced mesh sizes of the protein network, and a more regular distribution of the proteins in the yoghurt gel. As a result, yoghurt products from enzyme-treated milk showed increased gel strength and less syneresis, especially when the enzyme was not inactivated. Sensory studies revealed that odour and consistency properties of products from TG-treated milk were assessed as less 'yoghurt specific'. On the other hand, products from enzyme-treated milk were described as being more creamy, indicating that a TG treatment may simulate fat in fermented milk products.     

转谷氨酰胺酶对蛋白质凝胶性能的影响

本文研究了转谷氨酰胺酶浓度、反应温度、pH、时间对非肉蛋白质凝胶硬度的影响。结果表明转谷氨酰胺酶添加浓度在10-40U／g蛋白质，反应温度4-60℃，pH6-7，反应时间50-300min范围内，对于大豆分离蛋白、酪蛋白、乳清浓缩蛋白、卵清蛋白、明胶蛋白5种非肉蛋白质作用，均可以获得蛋白质凝胶能力以及凝胶性能的提高，表现在形成凝胶蛋白质极限浓度的降低以及凝胶硬度的提高。     

转谷氨酰胺酶对牛乳酪蛋白功能性质的影响

采用微生物转谷氨酰胺酶 (TG)对牛乳中的酪蛋白进行了酶法改性 ,并对改性后牛乳中酪蛋白功能性质如溶解性、起泡性、泡沫稳定性和持水性进行了研究 .结果表明 ,牛乳经TG作用后 ,其酪蛋白功能性质发生了较大变化 ,酪蛋白的起泡性、泡沫稳定性和持水性都有不同程度的增加 ,而溶解性略有下降 .实验中考察了TG作用的时间、温度、剂量等因素对酪蛋白功能性质的影响 ,结果显示 ,要获得综合性能优良的酪蛋白 ,TG最佳的作用条件为 :pH 7.0 ,5 0℃ ,6 0min ,酶的添加量0 .1～ 0 .2U/mL .     

添加谷氨酰胺转氨酶对凝固型酸奶品质的影响

研究添加谷氨酰胺转氨酶（简称TG酶）的凝固型酸奶与普通凝固型酸奶（未添加TG酶）在发酵过程中理化指标与感官指标的差异性。采用酚酞指示剂法、流变仪、质构仪、离心法和平板计数法分别测定酸奶在发酵过程中酸度、粘度、硬度、持水性及活菌数的变化;用电子眼、电子舌、电子鼻测定色泽、滋味、气味的变化,并进行感官评定。结果表明,每克乳蛋白添加0~5 U的TG酶,发酵结束时酸奶的酸度在71.12~73.64 oT范围内,酸度无显著性差异（P>0.05）;在发酵过程中,粘度、硬度均在3 h后呈显著上升（P<0.05）,发酵结束时,添加TG酶会显著增高凝固型酸奶的粘度和硬度（P<0.05）;随着TG酶添加量的增加,持水性显著增强（P<0.05）;添加不同浓度的TG酶凝固型酸奶中的活菌数无显著差异（P>0.05）。TG酶凝固型酸奶与普通凝固型酸奶的主色号均为4095,发酵结束时,色泽无显著性差异（P>0.05）;经主成分分析,发酵结束后添加不同浓度的TG酶不会对酸奶的滋味、气味造成显著性影响（P>0.05）;经感官评定,TG酶在添加量为2 U/（g蛋白）时感官品质最受喜爱。因此,与未加TG酶的普通凝固型酸奶相比,添加TG酶的凝固型酸奶在酸度、活菌数、色泽、滋味、气味方面均无显著性差异（P>0.05）;在粘度、硬度和持水性方面显著提高（P<0.05）,添加适量TG酶有利于凝固型酸奶品质的提高。     

Effect of Tetrasodium Pyrophosphate on the Physicochemical Properties of Yogurt Gels

The effect of tetrasodium pyrophosphate (TSPP) on the properties of yogurt gels was investigated. Various concentrations (0.05 to 0.2%) of TSPP were added to preheated (85°C for 30 min) reconstituted skim milk, which was readjusted to pH 6.50. Milk was inoculated with 2% starter culture and incubated at 42°C until the pH reached 4.6. Acid-base buffering profiles of milk and total and soluble calcium levels were measured. Turbidity measurements were used to indicate changes in casein dispersion. Storage modulus (G′) and loss tangent (LT) values of yogurts were monitored during fermentation using dynamic oscillatory rheology. Large deformation properties of gels were also measured. Microstructural properties of yogurt were observed using fluorescence microscopy. The addition of TSPP resulted in the disappearance of the buffering peak during acid titration at pH ∼5.1 that is due to the solubilization of colloidal calcium phosphate (CCP), and a new peak was observed at lower pH values (pH 4.0-4.5). The buffering peak at pH 6.0 during base titration virtually disappeared with addition of TSPP and a new peak appeared at pH ∼4.8. The addition of TSPP reduced the soluble Ca content of milk and increased casein-bound Ca values. The addition of up to 0.125% TSPP resulted in a reduction in turbidity because of micelle dispersion but at 0.15%, turbidity increased and these samples exhibited a time-dependent increase in turbidity because of aggregation of casein particles. Gels made with 0.20% TSPP were very weak and had a very high gelation pH (6.35), probably due to complete dispersion of the micelle structure in this sample. The LT value of gels at pH 5.1 decreased with an increase in TSPP concentration, probably due to the loss of CCP with the addition of TSPP. The G′ values at pH 4.6 of gels made with ≤0.10% TSPP were not significantly different but the addition of ≥0.125% TSPP significantly decreased G′ values. The addition of 0.05 to 0.125% TSPP to milk resulted in a reduction in the yield stress values of yogurt compared with yogurt made without TSPP. Greater TSPP levels (>0.125%) markedly reduced the yield stress values of yogurt. Lowest whey separation levels were observed in yogurts made with 0.10% TSPP. High TSPP levels (>0.10%) greatly increased the apparent pore size of gels. Addition of very low levels of TSPP to milk for yogurt manufacture may be useful in reducing the whey separation defect, but at TSPP concentrations ≥0.125% very weak gels were formed.     

培养基组成对轮枝链霉菌合成谷氨酰胺转胺酶的影响

对产谷氨酰胺转胺酶菌种轮枝链霉菌（Streptoverticillium）SK-1的摇瓶条件进行了研究,结果表明当起始pH值为6.5～8.0,氮源、碳源为蛋白胨与甘油,培养时间为110h时,酶活最高可达1.8μmol/(min     

微生物转谷氨酰胺酶催化酪蛋白酸钠聚合物的功能特性研究

研究了微生物转谷氨酰胺酶(TGase)催化酪蛋白酸钠聚合对其功能特性的影响。结果显示,随着催化时间的增加,其各组份含量不断减少,同时生成的聚合物含量不断增加,比较了TGase催化不同酪蛋白的聚合速率,结果显示转谷氨酰胺酶较易催化α-和β-酪蛋白,而不易催化κ-酪蛋白TGase催化聚合酪蛋白酸钠能明显改善其乳化性能,聚合时间越长,乳化指数越高,而乳化稳定性随聚合时间先升后降,4h时最高TGase催化酪蛋白酸钠还可提高其热稳定性,然而对其起泡性影响不大     

谷氨酰胺转氨酶对竹荚鱼鱼糜蛋白凝胶特性的影响

以质构、白度和持水力为指标研究谷氨酰胺转氨酶(TGase)添加量、凝胶化时间和凝胶化温度对竹荚鱼鱼糜凝胶特性的影响。结果表明：当TGase 添加量为80U/100g 鱼糜、凝胶化时间为5h、凝胶化温度为37.5℃时,竹荚鱼鱼糜的凝胶特性达到最佳,能够形成高度致密、均匀的凝胶网络结构。采用二段加热法制备的竹荚鱼鱼糜凝胶的特性好于一段加热法。     

Starch Phosphates Produced by Extrusion: Physical Properties and Influence on Yogurt Stability

Starch phosphorylation with sodium tripolyphosphate (STP) was performed using a single‐screw extrusion process. The extrusion variables studied were temperature (from 99.5°C to 200.4°C), water content (16.3% to 19.7%, w/w), and STP content (0.82% to 4.18%, w/w). The resulting phosphorylated starch was evaluated with respect to expansion, water solubility index (WSI), water absorption index (WAI), degree of substitution (DS), and viscosity. The phosphorylated starch was tested as a stabilizing additive in yogurt formulations, with respect to the syneresis index (SI) and the syneresis susceptibility coefficient (SSC). The results show that the expansion increases with increasing STP content, and decreases with increasing temperature and water content employed in the process. Increasing extrusion temperatures and decreasing water content resulted in increasing WSI and decreasing WAI. The DS was dependent on STP content, increasing as STP content increased, but appeared to be insensitive to changes in water content (between 16% and 20%, w/w). The highest level of yogurt stability, as measured by the syneresis index (SI) and the syneresis susceptibility coefficient (SSC), were obtained when the yogurt was formulated with the starch of the highest DS (0.018), which had been obtained at 150°C, 18% water content, and 4.12% (w/w) of STP.     

Effect of Epigallocatechin Gallate on the Properties of Gelatin

Epigallocatechin gallate was added to gelatin, and the changes in the gelatin were characterized to determine the effects of epigallocatechin gallate modification. The microstructural changes in the samples were analyzed using Fourier transform infrared spectroscopy, X-ray diffraction, differential scanning calorimetry, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and scanning electron microscopy. The results indicated that the gel strength and thermal stability of gelatin can be improved by appropriate epigallocatechin gallate addition. The optimal final concentration of epigallocatechin gallate was 1.0 g l^?1 in a gelatin solution (66.7 g l^?1). The concentration was also verified using Fourier transform infrared spectroscopy and X-ray diffraction. Covalent bonds were not observed in the epigallocatechin gallate-gelatin samples. Hydrogen bonds were the main molecular interactions observed in the epigallocatechin gallate-gelatin samples. The color of the epigallocatechin gallate-gelatin hydrogels or xerogels was darker because of the epigallocatechin gallate oxidation.     

转谷氨酰胺酶制剂对带鱼鱼糜制品质构特性的影响

研究了转谷氨酰胺酶制剂改善低值带鱼鱼糜制品质构特性的可行性。用氧肟酸-FeCl_3-三氯乙酸比色法测定了带鱼鱼糜原料和转谷氨酰胺酶制剂的酶活性。将不同百分比含量(0、0.3%、0.6%、0.9%)的转谷氨酰胺酶制荆分组添加到加水量分别为30%和60%的带鱼(Trichiunls haumela)鱼糜中,成型,反应一定时间(0℃,16h;40℃,1h),加热(85℃,40min),冷却至0～1℃,分别测定各组的质构特性(TPA)。结果表明:带鱼鱼糜原料具有一定的转谷氨酰胺酶活性0.110U/g,但活性较低。随着酶制剂添加量的增大,对照组与各处理组之间制品质构特性包括硬度、弹性均显著增加(p<0.05),而脆性变化不显著(P>0.05)。转谷氨酰胺酶制剂可有效改善低值鱼鱼糜制品质构特性。     

普通凝固型酸奶的工艺及品质控制

介绍了普通凝固型酸奶的制作工艺及质量控制的几个方面，通过正交试验，得到最佳工艺条件为：蔗糖8%，培养时间3h，培养温度44℃，接种量6mL。同时对产品的感官、理化及微生物指标进行了检验，评价了产品的品质情况，为普通凝固型酸奶的生产提供了工艺及质量控制的理论依据。     

Structural mechanisms leading to improved water retention in acid milk gels by use of transglutaminase

Water retention in transglutaminase (TG)-treated acid milk gels was studied and linked with the gel formation dynamics. Heat-treated skim milk with and without pre-treatment by TG was acidified at 20 °C, 30 °C and 40 °C at constant glucono-δ-lactone (GDL) level to obtain different acidification rates. Formation dynamics and structural properties of acid-induced gels were followed by rheological and near-infrared light backscattering measurements as well as microscopy. TG-treated gels showed decreased tan δ values all through the acidification, which was pronounced around the gelation point. Backscattered light intensity was lowered in TG-treated gels compared to the controls indicating that TG-treated gels were comprised of smaller aggregates. Water holding capacity (WHC) was measured by using centrifugation at selected pH points (pH 5.2, 5.0, 4.8 and 4.6) during acidification. Both acidification temperature and TG treatment had significant effects on the water retention properties of the gels. Spontaneous syneresis observed at high acidification temperatures (≥30 °C) was prevented upon TG-treatment. WHC of TG-treated gels was significantly higher compared to the control gels at all pH points. TG-treated milk gels showed a homogeneous network formed of smaller aggregate and pore sizes at the gelation point and did not show any large-scale re-organisation thereafter. Transglutaminase is likely to act as a fixative of the protein network at an early stage of gelation and thereby limiting network rearrangements that take place in acid milk gels formed at high acidification temperatures leading to contraction and subsequent wheying off.     

利用谷氨酰胺转胺酶生产大豆蛋白食用保鲜膜的研究

初步研究了大豆蛋白食用保鲜膜的成膜条件以及膜的透水性、透油性、水溶性等性能。试验结果表明,利用谷氨酰胺转胺酶生产的大豆蛋白食用保鲜膜,有较好的水蒸汽阻隔性能和隔油性,能达到食品保鲜的要求。     

谷氨酰胺转氨酶对大豆分离蛋白塑料拉伸性能的影响

研究了微生物谷氨酰氨转胺酶对大豆分离蛋白塑料拉伸性能的影响,发现在50℃时转氨酶对大豆分离蛋白催化聚合反应60 min,形成的高分子聚合物使大豆分离蛋白的抗拉强度和杨氏模量分别由未经处理时的9.2 MPa和265.4 MPa增加到14.7 MPa和390.1 MPa,同时增加了蛋白质的变性温度和热焓值。在酶处理90 min时,由于转氨酶变性失去活性及大豆分离蛋白的部分变性,蛋白质塑料的拉伸性能反而下降。对大豆分离蛋白的预热处理降低了转氨酶的催化聚合反应,且变性后的大豆分离蛋白制备的塑料拉伸性能大大降低。     

刺梨发酵乳的制备及其理化性质比较

刺梨含有丰富的多糖和维生素C 等营养成分以及超氧化物歧化酶（SOD）,具有降低慢性疾病风险及发病率的潜力。该研究选用保加利亚乳杆菌德氏乳杆菌DMLD-H1（Lactobacillus delbrueckii DMLD-H1,简称H1）和嗜热链球菌DMST-H2（Streptococcus thermophilus DMST-H2,简称H2）为发酵剂菌株,通过优化菌种比例、接种量和刺梨汁添加量,确定刺梨发酵乳最佳发酵工艺,并与商业菌种发酵的发酵乳的各项理化性质进行比较。实验表明,最佳的刺梨发酵乳发酵条件为：菌种比例为H1:H2=1:2,接种量为1.0×107 CFU/mL,刺梨添加量为0.06 g/mL。在此条件下发酵所得的刺梨发酵乳pH值为4.47,酸度为76.78 °T,持水力为32.94%,其中分离出11种挥发性物质,蛋白质相对分子质量主要分布在22~38 ku,后酸化4 周后发酵乳的pH值降低至3.89,活菌数含量为1.1×108 CFU/mL。对比商业发酵乳发现,刺梨发酵乳能够抑制发酵乳的后酸化作用,提高持水力,以及提升发酵乳中的酪蛋白含量;然而,刺梨可能会影响发酵乳中原本的风味物质。该研究为一种新型的刺梨发酵乳的开发提供理论依据,为新型功能性发酵乳的研发提供参考。     

发酵乳抗高血压特性的研究

本研究通过抑制ACE活性实验,筛选出能发酵脱脂牛乳产生较强抗ACE活性的LacobacillushelveticusATCC15019、15009和LactobacillusthermophilusATCC14485三种乳酸菌菌株,用这三种菌株混合制作的发酵乳对ACE的活性具有较强的抑制作用。分别给原发性高血压大鼠连续三天灌胃此发酵乳和其乳清,结果发现它们都具有较强的降高血压作用,最大降压值分别为24.10±2.69mmHg和29.14±3.07mmHg,与对照组相比,呈极显著性差异(p<0.01)。     

Study on the Functional Properties of Soybean Protein Isolate Cross-Linked with Gelatin by Microbial Transglutaminase

A microbial transglutaminase was used to cross-link soybean protein isolates in the presence of gelatin to prepare a cross-linked composite soybean protein-gelatin. Cross-linking conditions, such as total proteins, the ratio of soybean protein isolates to gelatin, and original pH, were fixed at 5% (w/v), 4:1 (w/w), and 7.5, respectively. The 4-hydroxyproline content in the modified product was measured and used as an index to select suitable transglutaminase addition, reaction temperature, and time by single factor experiments, which were found to be 20 U/g proteins, 40°C, and 2 h, respectively. Under the identified conditions, a modified product with 4-hydroxyproline content of about 36 mg/g proteins was prepared, and its functional properties were evaluated. SDS-PAGE analysis showed that several protein polymers existed in the modified product. Compared to the original soybean protein isolates or a cross-linked soybean protein isolates, the modified product had a better emulsifying stability and water holding capacity but a poor enzymatic digestibility in vitro, emulsifying activity and oil binding capacity; meanwhile, the dispersions formed by the modified product exhibited the highest apparent viscosity, storage modulus, and viscous modulus.     

Characteristics of enzymatically-deesterified pectin gels produced in the presence of monovalent ionic salts

Pectin methylesterases (PMEs) from Valencia orange (p-PME) and Aspergillus aculeatus (f-PME) were used to produce pectin gels in the presence of 0.2 M monovalent ionic salts. At pH 5.0, pectin gels were induced following enzymatic deesterification of high methoxy pectin, with greater deesterification observed using p-PME compared to f-PME. Under these conditions, the deesterification limit of f-PME ended up with a pectin of DE 30.5–31.9 which did not gel at the PME reaction completion, while p-PME reduced the pectin's DE to 16.0–17.2, resulting in gel formation. The pectin gel induced by KCl was significantly stronger than the NaCl-induced gel, but LiCl did not induce pectin gelation. The gel strength was influenced by both DE and species of monovalent cation. The KCl-induced gels released less water than NaCl-induced gels. A synergistic effect on gel strength was observed from the pectin treated with a combination of (p + f)-PMEs, producing even more stable gels. These results indicated that the pectin gelation of our system would be enhanced both by using larger monovalent cation and by lowering the DE value, which would presumably be attributed to the different action patterns recognized for p- and f-PMEs. This pectin gelation system could provide a useful alternative to acid-sugar or calcium cross-linked gels in food and other industrial applications.     

Characterization of fish gelatin from surimi processing wastes: Thermal analysis and effect of transglutaminase on gel properties

Fish gelatin extraction from wastes of fish Herring species (Tenualosa ilisha) was carried out by a series of pretreatment with 0.2 M Ca(OH)₂ followed by 0.1 M citric acid and final water extraction at 50 °C for 3 h. The resulting fish gelatin preparation was evaluated for its dynamic viscoelastic properties, gelling and melting temperatures and gel strength. The gelling and melting temperatures of gelatin samples (at 6.67%, w/v) were obtained from differential scanning calorimetry and rheological studies. The melting temperature of extracted fish gelatin (EFG) obtained ranged from 16.2 to 16.7 °C compared to that of commercial fish gelatin gel (CFG), from 23.7 to 25.6 °C and halal bovine gelatin (HBG), from 26.5 to 28.7 °C. On the other hand, gelling temperatures of EFG, CFG and HBG ranged from 5.1 to 5.2 °C, 11.9 to 17.46 °C, and 12.6 to 19.33 °C, respectively. EFG gave gels with a considerably lower G′ values than CFG and HBG. The bloom strength of EFG gels at 6.67% (w/v) was 69.03 g which was much lower than HBG (336.2 g) and CFG (435.9 g). Enzyme transglutaminase was added in the amounts of 0.5, 1.0, 3.0 and 5.0 mg/g gelatin to modify the gel properties of the extracted fish gelatin. The modified EFG gels obtained had higher gel strengths of 101.1 g and 90.56 g with added transglutaminase of 1.0 and 3.0 mg/g, respectively. However with addition of 5.0 mg/g enzyme the gel strength increased only up to 75.06 g. SDS-PAGE of extracted gelatin gel showed protein band intensities for α1-chains and 53 kDa but in gels added with higher concentration of transglutaminase, these protein band intensities seemed to disappear.