谷氨酰胺转胺酶的功能性质及在乳制品中的应用

介绍谷氨酰胺转胺酶的国内外研究现状,阐述其理化性质、来源及其在乳制品中的应用.特别是谷氨酰胺转胺酶在液态奶中对蛋白质、乳清蛋白及酪蛋白的影响和对酸奶品质的改良作用.     

谷氨酰胺转胺酶的特性及其应用研究进展

谷氨酰胺转胺酶是一种催化酰基转移反应的转移酶,它可改变蛋白的功能特性,使酪蛋白、肌球蛋白、谷蛋白等分子间和分子内产生共价交联。介绍了谷氨酰胺转胺酶的来源、性质、作用机理和在食品、纺织、医药及皮革中的应用前景。     

婴儿配方乳蛋白水解物在酶-膜反应器中连续化生产试验研究

本试验首先通过调整牛乳中酪蛋白与乳清蛋白的比例为40:60,然后使其在连续搅拌式酶-膜反应器(continuousstirredtankmembranereactor,CSTMR)中水解以生产婴儿配方乳的基料。通过对三种蛋白水解酶的比较筛选最终确定以胰蛋白酶为水解用酶;单因素试验确定模拟母乳在CSTMR中水解的最适温度40℃、最适pH8.0;通过正交试验得出乳蛋白在CSTMR中水解的最适酶底比(E/S,W/W)1.84%、最适底物浓度(S,W/V)4%、最适压力(P,MPa)为0.05MPa;对乳蛋白水解物做SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测,发现乳蛋白水解液中酪蛋白的谱带较水解之前明显减轻,电泳凝胶扫描分析结果显示,酪蛋白中的αs-酪蛋白几乎被水解掉90%,乳清中的β-乳球蛋白也有50%以上的降解。     

枯草芽孢杆菌谷氨酰胺转胺酶基因的克隆及在大肠杆菌中的表达

以枯草芽孢杆菌DB104染色体DNA为模板,利用PCR技术扩增得到谷氨酰胺转胺酶基因tg,将其克隆至大肠杆菌质粒pET-22b( )中,构建表达栽体pET-tg,并转化大肠杆菌BL21(DE3).重组菌经IPTG(异丙基β-D-硫代半乳糖苷)诱导,SDS-PAGE电泳分析,结果表明谷氨酰胺转胺酶得到了表达,表达量占菌体总蛋白的12%.诱导菌体经裂解后的上清与酪蛋白反应,显示其具有交联蛋白质的活性.经荧光法测定,诱导5 h的菌体酶活为1 590 U/g(DCW,cell weight,细胞干重),是出发菌株30 U/g(DCW)的52倍.     

重组牛肉生产牛肉干的研究

谷氨酰胺转胺酶是一种催化酰基转移反应的转移酶,它可使用酪蛋白、肌球蛋白、谷蛋白和乳球蛋白等分子之间产生交联,改变蛋白质的功能性质。采用碎牛肉为主要原料,配以各种辅料,添加谷氨酰胺转胺酶,经过多次试验,优化工艺配方,研制出一种色泽、口感、风味均被人们接受的重组牛肉干。     

微生物谷氨酰胺转胺酶的应用进展

谷氨酰胺转胺酶是一种催化酰基转移反应的转移酶,它可使酪蛋白、肌球蛋白、谷蛋白和乳球蛋白等分子之间产生交联,改变蛋白质的功能性质,简述了谷氨酰胺转胺酶在食品应用的一些最新进展。     

谷氨酰胺转胺酶的发酵生产和应用研究

谷氨酰胺转胺酶催化体外大多数食品蛋白质的交联反应,如：酪蛋白,大豆蛋白,肌球蛋白,肌动蛋白,谷蛋白,禽蛋蛋白等等。通过催化肽键谷酰胺基残基的酰基转移反应,在各种蛋白质分子之间或之内形成ε-（γ－谷胺酰）赖氨酸键,从而改善各种蛋白质的功能性质。本文就谷氨酰胺转胺酶的国内外研究现状作一综述,主要包括理化性质,生产工艺,提取技术及其在食品工业中的应用。另外对本研究室谷氨酰胺转胺酶发酵法生产的研究情况也作了报道。     

谷氨酰胺转胺酶在畜产加工中的应用

谷氨酰胺转胺酶是一种催化酰基转移反应的酶，能在蛋白质分子间或分子内生成ε-（γ－谷氨酰）赖氨酰共价键，而形成强有力的凝胶，从而改善各种蛋白制品的弹性、粘合性、保水性等品质。谷氨酰胺转胺酶能以各种动物性蛋白作为反应底物（如肌球蛋白，肌动蛋白，酪蛋白等），在畜产加工中有广泛的应用。     

谷氨酰胺转胺酶的性质及在食品中的应用

谷氨酰胺转胺酶是一种催化酰基转移反应的转移酶,它可使酪蛋白、肌球蛋白、谷蛋白和乳球蛋白等蛋白质分子之间产生交联,从而改变食品蛋白质的功能性质。本论述了谷氨酰胺转胺酶的性质及在食品开发中的应用。     

牛乳中αs、β-酪蛋白组分的分离

以新鲜脱脂牛乳为原料,在碱性条件下添加钙盐进行选择性沉淀分离αs-、β- 酪蛋白组分,并用十二烷基磺酸钠- 聚丙烯酰胺凝胶电泳(SDS-PAGE)法测定分离效果。从CaCl2 溶液终浓度、冷却温度和反复溶解﹑沉淀次数三个方面对αs-、β- 酪蛋白的分离效果进行研究。结果表明：CaCl2 溶液终浓度0.065mol/L,冷却温度2℃,经3 次反复溶解﹑沉淀,分离得到的αs- 酪蛋白组分纯度较高,可达83.33%,β- 酪蛋白组分纯度为109.53%。     

Study of the shear and extensional rheology of casein,waxy maize starch and their mixtures

This work investigated the shear and uniaxial extensional flow behaviour of aqueous casein and phosphated waxy maize starch systems as a function of the deformation rate, biopolymer concentration and the temperature. Trouton ratios were calculated to compare different responses of biopolymers to the shear and extensional deformation. It was found that the casein system (20% w/w) had a much higher and linearly increasing Trouton ratio against the log increase of strain rate. The starch system (35% w/w) had a Trouton ratio close to 3 at low strain rates, but increasing linearly once the strain rate exceeded 2 s⁻¹. The apparent shear and extensional viscosity showed an exponential increase with the concentration for both biopolymers, with casein being more concentration dependent. Their Trouton ratios were also very much concentration dependent: remained close to 3 at low biopolymer levels, but increased significantly at higher concentrations. Temperature variation experiments demonstrated that the flow properties of casein and waxy maize starch follow an Arrhenius relationship, with casein showing stronger temperature dependence than starch. While casein systems displayed a decrease in Trouton ratio with temperature increase, waxy maize starch had the opposite behaviour. The mixing of casein–waxy maize starch resulted in higher apparent extensional viscosities and higher Trouton ratios than single biopolymer systems.     

Soybean and milk proteins modified by transglutaminase improves chicken sausage texture even at reduced levels of phosphate

Consumer demands for poultry processed meats have increased due to low fat content. In this experiment, chicken sausages were manufactured with various biopolymers prepared from soybean protein, casein, whey protein isolate (WPI), mixtures of soybean protein and casein, and soybean protein and WPI. The extent of various biopolymer formations was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high performance liquid chromatography. Cross-linking soybean protein and casein or WPI by transglutaminase provided biopolymers with improved heat stability and emulsifying property. Shear force of chicken sausages were measured to evaluate the addition of biopolymer on the hardness in the presence of 0.05 or 0.2% sodium tripolyphosphate (STPP). The texture of chicken sausages was improved by the addition of such biopolymers even in the presence of 0.05% STPP. These results suggested that chicken sausage texture was improved by the formation of network structures that contribute to hardness of sausage gels with the addition of biopolymers. Thus, addition of biopolymers in the manufacture of chicken sausages may permit reduction in phosphate content without loss in texture.     

自组装原花青素纳米复合物的构建及表征

原花青素是一种具有极强抗氧化活性的天然多酚化合物,该文利用酪蛋白和麦芽糊精进行分子自组装得到一种两亲性生物聚合物,将其与原花青素结合形成纳米复合物。通过接枝度、褐变度、蛋白溶解度以及红外光谱和圆二色谱分析酪蛋白与麦芽糊精之间的相互作用机制。以包埋率为指标,优化原花青素纳米复合物构建的工艺条件,对原花青素纳米复合物的粒径、电位和多分散指数进行表征,并采用扫描电镜观察其包埋前后表观的形貌变化。结果表明,酪蛋白与麦芽糊精成功地进行自组装反应,该反应有效地提高了酪蛋白的溶解度。当原花青素/接枝物的质量比为2∶5,接枝物浓度为25 mg/mL,溶液pH值为7.0时,制备得到的原花青素纳米复合物包埋率达到93.48%,平均粒径为158.69 nm,Zeta电位为-30.58 mV,呈现一种表面光滑的紧凑球状结构。     

转谷氨酰胺酶催化大豆蛋白和乳清蛋白合成耐热性聚合蛋白

用商品级转谷氨酰胺酶(TG-B)聚合大豆蛋白和乳清蛋白形成高耐热、耐酸的蛋白聚合物。蛋白聚合物的合成量由SDS-PAGE电泳结合凝胶成像分析测定;蛋白聚合物的耐热性用差示扫描量热法(DSC)测定;蛋白聚合物的酸溶解性用双缩脲法测定。结果表明TG-B聚合大豆蛋白和乳清蛋白形成的蛋白聚合物的最适条件为pH为6～7;反应温度30℃～45℃,反应时间4h,加酶量为6当量单位/g蛋白,在此条件下蛋白聚合物的转化量可达30%,所合成蛋白聚合物可耐130℃的热处理而不发生变性;并在pH3.2～4.3范围不发生沉淀。     

Formation and potential uses of milk proteins as nano delivery vehicles for nutraceuticals: A review

The aim of this review was to highlight the progress achieved in the use of milk protein as nano vehicles for nutraceuticals. Reassembled casein and β‐casein micelles and core/shell nanoparticles from casein with other biopolymers have been prepared. Also, cross‐linking of casein micelles has developed stable nanoparticles. Nanogels of whey proteins (WP), β‐lactoglobulin (β‐LG) and lactoferrin (Lf) have been prepared by controlled thermal treatment, and several core/shell nanoparticles have been developed from WP or β‐LG with several polysaccharides. The developed caseins and WP nanoparticles have been used as carriers for several nutraceuticals. Examples have been presented and discussed.     

Cross-Linking and Glucosamine Conjugation of Casein by Transglutaminase and the Emulsifying Property and Digestibility In Vitro of the Modified Product

Microbial transglutaminase was applied as a biocatalyst and glucosamine as an acyl acceptor to modify casein by cross-linking and glucosamine conjugation. Electrophoretic analysis revealed that transglutaminase-induced cross-linking and glucosamine conjugation occurred simultaneously during reaction, and some polymers of glycoproteins with higher molecular weights were formed in the modified casein product prepared. HPLC analysis demonstrated that about 1.2 mol of glucosamine was conjugated to 1 mol of casein, under the preparation conditions as follows: casein concentration of 3% (w/v), molar ratio of acyl donor in casein to glucosamine acceptor of 1:3, transglutaminase addition level of 10 U/g casein, reaction temperature of 37°C, and reaction time of 4 h. The evaluation results also showed that the surface hydrophobicity of the modified casein product decreased. The emulsifying activity index and emulsifying stability index of the modified casein product were 100.9 m²/g and 84.3%, about 12 and 20% higher than that of original casein. The digestibility in vitro of the modified casein product was the same as that of original casein or had an increase of 9.8%. Meanwhile, cross-linking of casein by transglutaminase in the absence of glucosamine showed adverse impact on the emulsifying properties of the cross-linked casein prepared. The new modification method developed might have the potential as an effective approach to improve the functional properties of food proteins.     

Properties of Biopolymers from Cross-linking Whey Protein Isolate and Soybean 11S Globulin

Biopolymers were prepared by cross-linking whey protein isolate (WPI) and soybean 11S using transglutaminase. Electrophoretic pattern, solubility, emulsification, hydrophobicity and foaming properties of the biopolymers were determined. SDS-PAGE showed bands corresponding to high-molecular-weight components (MW>200 kDa). Biopolymer solubility was > 90% at pH 3 and below, and at pH 7 and above. Emulsifying properties of biopolymers were lower than those of WPI. The foaming capacity of the biopolymers (23.6 mL) and WPI/11S mixture (22.7 mL) were similar to that of egg albumin (20.3 mL). The foaming stability of the biopolymers (122 min) was higher than that of WPI/11S mixture (33.7 min), and was similar to that of egg albumin (132 min).     

The association of lipophilic phospholipids with native bovine casein micelles in skim milk: Effect of lactation stage and casein micelle size

A known biological role of casein micelles is to transport calcium from mother to young and provide amino acids for growth and development. Previous reports demonstrated that modified casein micelles can be used to transport and deliver hydrophobic probes. In this study, the distribution of lipid-soluble phospholipids, including sphingomyelins (SM) and phosphatidylcholines (PC), was quantified in whole raw milk, skim raw milk, and casein micelles of various sizes during early, mid, and late lactation stages. Low-pressure size exclusion chromatography was used to separate casein micelles by size, followed by hydrophobic extraction and liquid chromatography–mass spectrometry for the quantification of PC and SM. Results showed that the SM d18:1/23:0, d18:1/22:0, d18:1/16:0, d16:1/22:0, d16:1/23:0, and d18:1/24:0 and the PC 16:0/18:1, 18:0/18:2, and 16:0/16:0 were dominating candidates appearing in maximum concentration in whole raw milk obtained from late lactation, with 21 to 50% of total SM and 16 to 35% of total PC appearing in skim milk. Of the total SM and PC found in skim milk, 35 to 46% of SM and 22 to 29% of PC were associated with the casein micelle fraction. The highest concentrations of SM d18:1/22:0 (341 ± 17 µg/g of casein protein) and PC 16:0/18:1 (180 ± 20 µg/g of casein protein) were found to be associated with the largest casein micelles (diameter = 149 nm) isolated in milk from late lactation, followed by a decrease in concentration as the casein micelle size decreased.     

MTGase聚合酪蛋白酸钠生物聚合物的结构特征研究

采用紫外光谱、荧光光谱及红外光谱分析技术,研究了微生物转谷氨酰胺酶(MTGase)聚合酪蛋白酸钠(Na-CN)生物聚合物的空间结构特征,并探讨了MTGase改善Na-CN乳化性能的作用机理。紫外光谱显示,MTGase聚合Na-CN生物聚合物的多肽链的Trp和Tyr残基的紫外吸收峰的强度明显低于Na-CN,说明生物聚合物的“空间结构效应”占较重要的地位。荧光发射光谱显示,Na-CN生物聚合物的Trp和Tyr残基的荧光强度比Na-CN有显著的增强,表明生物聚合物的疏水性区域更加暴露。然而,MTGase长时间催化(12h)得到的生物聚合物的荧光强度反而有所下降(与4h的场合相比),这反映了“空间位阻效应”。红外光谱显示,Na-CN与其生物聚合物的酰胺特征峰相差不大,说明两者的二级结构基本上相近。此外,MTGase改善Na-CN乳化性能的机理是:MTGase催化导致Na-CN的空间结构发生了变化,进而改变了蛋白表面的表面疏水性质,最终达到改善Na-CN乳化性质的效果。     

基于酪蛋白自组装法制备α-生育酚/酪蛋白纳米粒及其α-生育酚稳定性分析

采用自组装法使酪蛋白与α-生育酚形成α-生育酚/酪蛋白纳米粒并考察其稳定性。以纳米颗粒的荧光强度为指标,考察在单因素试验条件下α-生育酚与酪蛋白的结合强度。对所制备的复合纳米粒表征后以α-生育酚保留率为指标进行稳定性测试。结果表明制备纳米复合物最佳条件为组装温度37?℃、pH?6.8、α-生育酚与酪蛋白质量比1∶300,所得α-生育酚/酪蛋白纳米粒平均粒径为（135.6±13.7）nm,包封率为（97.97±7.38）%,载药量为（0.33±0.03）%。红外光谱表明α-生育酚与酪蛋白存在较强氢键作用。电镜观察纳米粒形态学结构大小均匀,呈较规则球形,且分散性较好。稳定性测试表明复合纳米粒贮存稳定性、热稳定性、冻干稳定性及抗氧化性良好。该方法不仅保护了α-生育酚,而且扩大了产品的应用范围,使其在食品加工中具有理想的应用前景。