谷氨酰胺转胺酶在搅拌型酸奶加工中的应用

主要研究了谷氨酰胺转胺酶在搅拌型酸奶加工中的应用,研究结果表明:该酶的最适添加顺序为与发酵剂同时加入到灭菌的乳中;最适添加种类为TG-K;最适添加量为3×10-4g/mL;而且还研究了在贮藏期间,加酶酸奶的质构变化情况。     

Mozzarella干酪内聚性的优化

研究TGase、凝乳酶、CaCl2对Mozzarella干酪内聚性的影响,以优化工艺参数。采用三因素二次正交回归设计确定TGase、凝乳酶、CaCl2的添加量水平,通过SAS软件RSREG过程,分析三因素主效应及其交互效应对成熟10 d的干酪内聚性的影响。结果表明,三因素对内聚性的影响符合Y=B0+∑BjXj+∑BijXiXj+∑BjjX2的三元二次回归模型,其关键参数主效应为TGase(P<0.05)>凝乳酶(P<0.05)>CaCl(2P<0.05);TGase、凝乳酶和CaCl2对干酪内聚性有明显的交互作用。最优工艺参数为:TGase 1.046%,凝乳酶0.817%,CaCl20.075 g/L,内聚性值最大0.628。     

赖氨酸与谷氨酰胺在谷氨酰胺转胺酶作用下的交联反应

本文研究了赖氨酸与谷氨酰胺在谷氨酰胺转胺酶催化下的交联反应。探讨了底物比、加酶量、pH值、反应温度和反应时间对聚合度的影响。利用Design-Expert6.0.3软件优化得出交联反应的最佳工艺条件为底物比2.15、加酶量14.39U/g、pH值8.0、温度42℃和反应时间94min。同时,分析了各因素之间的交互效应。     

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.     

Comparison of Atlantic menhaden gels from surimi processed by acid or alkaline solubilization

Heat-induced gelling abilities of surimis prepared by pH shifting (isoelectric precipitation following acid (AC) or alkaline (AL) solubilization) were compared to that of conventionally washed (CW) surimi. Greater endogenous transglutaminase activity (evidenced as enhanced strength of cooked gels subjected to 30–40 °C preincubation) was measured for CW and AL surimi than for AC surimi (all at pH 7). Upon addition of microbial transglutaminase (MTGase), increased crosslinking of myosin heavy chain and gel strengthening during 30–40 °C preincubation were apparent for all three types of surimi, most markedly in CW and AL surimi. Salt addition improved CW gels most, but seemed to adversely affect MTGase activity in AC and AL surimi. AC and AL surimi gels were of lower whiteness than were CW surimi gels.     

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。     

微生物转谷氨酰胺酶(MTGase)的蛋白质底物催化特性及其催化机理研究 (I)MTGase催化单底物蛋白质的聚合特性

采用SDS-PAGE结合凝胶成像技术比较了MTGase在还原状态下对酪蛋白酸钠(SC)、牛血清蛋白(BSA)、大豆球蛋白(glycinin)和β-伴豆球蛋白(β-conglycinin)、β-乳球蛋白(β-LG)和α-乳白蛋白(α-LA)等单底物蛋白的聚合效率。结果表明MTGase较易催化SC和BSA聚合,其次为大豆球蛋白,而β-伴豆球蛋白、β-LG和α-LA最不易。根据MTGase催化不同单底物蛋白质的聚合速率的差异,对MTGase催化单底物蛋白质的聚合特性进行了探讨,指出:①底物蛋白的分子结构对MTGase催化活性的重要性;②蛋白质表面疏水度对MTGase催化活性的重要性;③对蛋白质进行一定的预处理可增强MTGase对蛋白质(特别是球蛋白)的催化活性。     

Effect of reducing agents on physicochemical properties and gel-forming ability of surimi produced from frozen fish

Effects of different reducing agents (cysteine, ascorbic acid and sodium bisulfite) at various levels on physicochemical properties of protein, transglutaminase activity and gel properties of surimi produced from frozen croaker, lizardfish, threadfin bream and bigeye snapper were studied. Addition of cysteine resulted in the highest increase in the breaking force and the deformation of surimi gels, compared with other reducing agents. The optimum levels of cysteine were 0.05, 0.1, 0.05 and 0.1% (w/w) for surimi from frozen croaker, lizardfish, threadfin bream and bigeye snapper, respectively. Surimi from frozen croaker with cysteine added showed a similar breaking force and deformation to that produced from fresh fish. With addition of cysteine, an increase in sulfhydryl content with a concomitant decrease in disulfide bond content was generally observed. Ca²⁺ ATPase activity also increased, indicating the renaturation of the myosin molecule. T_max of peak 1 (myosin peak) of all surimi sols in the presence of cysteine was shifted to higher temperature. The increased transglutaminase activity was observed with addition of cysteine. Therefore, reducing agents, especially cysteine, recovered the denatured muscle proteins and activated the transglutaminase in the muscle, leading to the increased gel-forming ability of surimi produced from frozen fish.     

Physical properties of yoghurt manufactured with milk whey and transglutaminase

The effect of milk protein polymerization prior to the yoghurt fermentation process was evaluated by enzymatic reaction with microbial transglutaminase (Streptoverticillium mobaraense). Yoghurt samples were manufactured with 100% milk or by substituting milk with 20 or 30% of liquid milk whey, aimed at determining the use of natural milk whey in dairy products. Transglutaminase was added at a protein ratio of 0.5 U g⁻¹ to all samples and evaluated regarding rheological behavior, syneresis index and texture profile. The addition of enzyme transglutaminase contributed to syneresis prevention and increased the consistency index in yoghurt samples manufactured with milk whey. Yoghurt manufactured from 70% milk plus 20% milk whey, followed by enzymatic treatment, presented similar characteristics to traditionally manufactured yoghurt (C 100), with no alteration in the syneresis of the samples (p > 0.05) and presented texture parameters similar to the control yoghurt (C 100).     

Effect of chymotrypsin digestion followed by polysaccharide conjugation or transglutaminase treatment on functional properties of millet proteins

Millet protein was solubilized by chymotrypsin; the soluble protein was conjugated to galactomannan under controlled conditions (60 °C, 76% RH) or polymerised by transglutaminase (TGase). SDS–PAGE patterns showed that the conjugated and polymerised proteins had higher molecular mass bands above the stacking gel. SDS–PAGE patterns also indicated that the digest was conjugated to galactomannan and polymerised by TGase. The free amino groups (OD₃₄₀) of the conjugated and polymerised digest were greatly reduced. Although the chymotrypsin digest was considerably insoluble between pH 2.0 and 5.0, galactomannan conjugate was completely soluble at all levels of pH. TGase polymer was slightly insoluble at pH 4.0. Galactomannan conjugate resisted heat-induced aggregation, even after heating at 90 °C for 20 min, while TGase polymer resisted heat-induced aggregation up to 70 °C, after which its solubility started to decline. The emulsifying properties of the conjugate and the polymerized proteins were greatly improved, compared to the native and chymotrypsin digests.