应用二次回归正交旋转组合设计优化转谷氨酰胺酶作用下的乳清蛋白交联条件

考察了pH、催化时间、催化温度及加酶量对转谷氨酰胺酶作用WPI形成交联产物的粘度影响,并对影响因素进行优化,找出最佳组合。该研究共分两个实验进行,实验1考察pH、时间、温度及加酶量单个因素对转谷氨酰胺酶交联作用的影响;实验2是基于单因素实验结果,采用四因素(pH、催化时间、催化温度、加酶量)五水平回归正交旋转设计,对WPI经转谷氨酰胺酶交联后产生最大粘度的最佳条件进行优化,以便确定实验多元回归方程和获得较大WPI黏度的最佳条件。实验结果表明:交联时间4h、交联温度50℃、pH8.0和加酶量20u/g时,具有最佳粘度值,转谷氨酰胺酶作用效果最佳。      

应用二次回归正交旋转组合设计优化黄霉素发酵培养基

在摇瓶条件下，采用二次回归正交旋转组合设计，优化黄霉素发酵培养基各组份的配比。优化结果表明：在葡萄糖、淀粉、硫酸铵、硫酸镁、碳酸钙的浓度为1％、3．75％、0．25％、0．015％、0．3％时，发酵效价最高。     

二次正交旋转组合设计法优化咸蛋清酶解条件

在木瓜蛋白酶酶解咸蛋清的单因素实验基础上,采用二次正交旋转组合设计实验方法对酶解工艺进行了优化。建立了底物浓度（X1）、加酶量（X2）、温度（X3）、pH（X4）与水解度（Y）的经验数学模型:Y=25.77583-4.01800X1+1.42650X3-1.48800X4-1.52933X21-2.50133X22-3.44708X23-3.02933X24+1.05450X1X2-1.58850X2X3。该模型的R2=96.49%,R2adj=94.14%,说明回归方程对实际拟合情况较好,自变量和响应值之间线性关系显著。最终得到咸蛋清酶解最佳条件为:底物浓度3%、加酶量5000U/g、酶解温度50℃、pH5.5、酶解6h。最佳条件下酶解咸蛋清水解度高达26.18%;氮收率可达93.47%。      

二次旋转正交组合设计优化羊骨蛋白酶解工艺

以新鲜羊骨为原料,选用碱性蛋白酶和风味蛋白酶按序分步水解羊骨蛋白,采用二次旋转正交组合设计优化双酶水解羊骨蛋白工艺条件。结果表明,在初始pH为8.5,初始温度为55℃,羊骨蛋白的最佳酶解方案为：底物浓度为10%,碱性蛋白酶添加量为3000u/g,作用时间为5h;风味蛋白酶添加量为3500u/g,作用时间为3.5h。     

二次正交旋转组合设计法优化咸蛋清酶解条件

在木瓜蛋白酶酶解咸蛋清的单因素实验基础上,采用二次正交旋转组合设计实验方法对酶解工艺进行了优化。建立了底物浓度(X1)、加酶量(X2)、温度(X3)、pH(X4)与水解度(Y)的经验数学模型:Y=25.77583-4.01800X1+1.42650X3-1.48800X4-1.52933X21-2.50133X22-3.44708X23-3.02933X24+1.05450X1X2-1.58850X2X3。该模型的R2=96.49%,R2adj=94.14%,说明回归方程对实际拟合情况较好,自变量和响应值之间线性关系显著。最终得到咸蛋清酶解最佳条件为:底物浓度3%、加酶量5000U/g、酶解温度50℃、pH5.5、酶解6h。最佳条件下酶解咸蛋清水解度高达26.18%;氮收率可达93.47%。     

应用二次回归正交旋转组合设计优化中国毛虾蒸煮液酶解工艺的研究

以蛋白水解度为指标,在中国毛虾蒸煮液单因素酶解研究的基础上,采用二次回归正交旋转组合设计对其酶解工艺进行优化.建立了蛋白水解度与酶解初始pH、酶解时间及蛋白酶用量三个实验因素的正交回归模型方程,根据回归模型进行主效应分析,通过频率分析法得到了酶解最佳工艺条件:酶解初始pH为7.6,酶解时间为8h,蛋白酶用量为0.51%(w/w),最佳条件下的水解度为38.8%.     

应用二次回归正交旋转组合设计优化中国毛虾蒸煮液酶解工艺的研究

以蛋白水解度为指标,在中国毛虾蒸煮液单因素酶解研究的基础上,采用二次回归正交旋转组合设计对其酶解工艺进行优化。建立了蛋白水解度与酶解初始pH、酶解时间及蛋白酶用量三个实验因素的正交回归模型方程,根据回归模型进行主效应分析,通过频率分析法得到了酶解最佳工艺条件:酶解初始pH为7·6,酶解时间为8h,蛋白酶用量为0·51%(w/w),最佳条件下的水解度为38·8%。      

应用二次回归正交旋转组合设计优化五倍子多糖提取工艺

考察时间、温度、料液比、粉碎度及超声功率五个单因素对超声波提取五倍子多糖的影响，并采用二次回归正交旋转组合设计优化多糖提取率的最佳提取时间、料液比、超声功率条件的试验回归方程，确定超声波提取五倍子多糖的最佳工艺条件为：提取时间30min，料液比1：20，超声功率300W。     

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。     

二次回归正交旋转组合优化罗非鱼下脚料酶解液螯合锌的工艺条件

以罗非鱼下脚料酶解液为原料,在单因素的基础上,用二次回归正交旋转组合方法优化其与硫酸锌的螯合条件,结果显示,在配位比2.70∶1、温度71.3℃、时间15min、pH6.86条件下,酶解液与硫酸锌的螯合率可达到87.56％ 各个因素对螯合率影响大小的顺序是:配位比＞pH＞温度＞时间.     

二次通用旋转组合设计法优化酶法提取茶叶籽蛋白工艺

采用二次通用旋转组合设计实验优化碱性蛋白酶提取茶叶籽蛋白工艺。在单因素实验基础上,选择碱性蛋白酶用量、pH、酶反应时间与温度为实验因素,以茶叶籽蛋白提取率为指标建立回归数学模型。对实验结果进行方差分析及对数学模型进行优化,获得了茶叶籽蛋白酶法提取的最佳条件为:料液比1:25、碱性蛋白酶用量200U/g、pH10.5、酶反应时间55min、酶反应温度45℃,在此条件下蛋白提取率可达75.83%。      

Incorporated glucosamine adversely affects the emulsifying properties of whey protein isolate polymerized by transglutaminase

Glucosamine (GlcN) and microbial transglutaminase (Tgase) are used separately or together to improve the emulsifying properties of whey protein isolate (WPI). However, little is known about how the emulsifying properties change when GlcN residues are incorporated into WPI cross-linked by Tgase. We used Tgase as a biocatalyst to cross-link WPI in the presence of GlcN in a liquid system for 12 h at a moderate temperature (25°C). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analyses indicated that protein polymerization and GlcN conjugation occurred simultaneously, phenomena also supported by the loss of free amines (9.4–20.5%). Addition of 5 U Tgase/g protein improved the emulsifying properties of moderately cross-linked WPI polymers. The Tgase-treated WPI polymers had a larger particle size (～2.6-fold) than native WPI, which may have reduced coalescence and flocculation in an oil-in-water emulsion system. However, the incorporation of GlcN residues into WPI, predominantly via enzymatic glycation, partly inhibited the cross-links between the WPI molecules catalyzed by Tgase, reducing the size of the WPI polymers 0.81- to 0.86-fold). Consequently, WPI+GlcN conjugates provided less stability to the emulsion. Moreover, high NaCl concentration (0.2 M) decreased the emulsifying properties of the WPI+GlcN conjugates by neutralizing negative electric charges in the glycoconjugates. However, the improved emulsifying properties of WPI cross-linked by Tgase may be useful in food processing at higher NaCl concentrations due to the formation of the thicker steric barrier at the oil-water interface.     

微生物谷氨酰胺转胺酶对乳清蛋白的改性

介绍了该酶的来源、性质及催化反应机理。由于乳清蛋白应用于食品加工中时，其理化功能尚不突出，所以通过谷氨酰胺转胺酶对乳清蛋白的改性，可以加强乳清蛋白的功能性质，从而合理利用资源，开发新产品，扩大其在食品中的应用。     

微生物转谷氨酰胺酶催化乳清蛋白聚合研究

采用SDS－PAGE分析，研究了不同条件下微生物转谷氨酰胺酶（MTGase）催化乳清蛋白（WPI）聚合。结果显示，MTGase可催化乳清蛋白的β－乳球蛋白（β-LG）和α－乳清蛋白（α－LA）聚合，形成低聚物或生物聚合物，其中β-LG更易受MTGase的催化，当TGase酶浓度一定时（0.5U/mL),TGase催化WPI聚合的最佳底物质量分数范围为2％－4％，对WPI进行加热预处理，同时添加还原剂，可明显提高MTGase对WPI的催化活性，MTGase催化WIP的最适PH值范围为6．5－7．5，当WPI经预热处理（85℃，15min），同时添加20mmol/L的DTT，TGase催化WPI聚合12h，可使质量分数为92％的β-LG和质量分数为75％的α－LA聚合。     

二次正交旋转组合设计优化香菇废菌棒蛋白质的提取工艺

采用二次正交旋转组合设计对香菇废菌棒蛋白质提取工艺进行优化。在单因素实验基础上,选择碱浓度、浸提温度、浸提时间、液固比为自变量,香菇废菌棒蛋白质提取量为检测指标,利用二次正交旋转组合进行四因素五水平的实验设计,并做统计分析,建立数学回归模型。结果表明,所得回归方程达到极显著水平,无失拟因素存在。蛋白质的最佳提取工艺条件为:碱浓度0.11mol/L,浸提温度72℃,浸提时间104min,液固比47:1,在此条件下,香菇废菌棒蛋白质提取量理论值为2.38mg/g,验证实测值为2.42mg/g,与理论值相对误差为0.04mg/g,说明所优化的工艺参数准确可靠、稳定、可行。      

Expanded functionality of modified whey protein dispersions after transglutaminase catalysis

The functionality of whey dispersions, prepared with a modified whey protein concentrate (mWPC) ingredient, was significantly altered after cross-linking with microbial transglutaminase (TGase) upon pH adjustment to 8. Test TGase-mWPC solutions, pH 8, gelled faster than control mWPC dispersions, as measured in real time; whereas, the gelling temperature of pretreated TGase-mWPC samples (37 °C, 2.5 h) increased from 67.8 to 74.8 °C with a minimal change in gel strength. Prolonged prior incubation with the enzyme (37 °C, 20 h) raised the gel strength in both control mWPC and TGase-mWPC dispersions, though these values were approximately 2.7 times lower in TGase-mWPC samples. Furthermore, the gelling temperature was raised by 9 °C after extensive polymerization. The water holding capacity was not impacted by enzymatic processing while emulsions prepared with TGase-mWPC dispersions proved very stable with no evidence of phase separation during storage at room temperature for 1 mo. Moreover, the apparent viscosity of TGase-mWPC emulsions exhibited a 10-fold increase compared to nonenzyme-treated mWPC samples. The particle size was nearly 11 μm in covalently linked TGase-mWPC test fractions compared with 8 μm in nonpolymerized mWPC dispersions. Ultimately, the functional characteristics of TGase-mWPC ingredients may be designed to deliver superior performance, especially with regard to improving heat and emulsion stability.     

乳清蛋白在转谷氨酰胺酶作用下包埋益生菌的研究

为了提高双歧杆菌在人体胃肠道中的存活率,以乳清蛋白为壁材,转谷氨酰胺酶为交联剂,通过乳化凝胶的方法制备包埋有两歧双歧杆菌的蛋白质微球。实验表明:以此工艺制备的微球成球性较好,粒径为(308.2±16.2)μm,益生菌包埋率为87.8%±10.0%,与未包埋的两歧双歧杆菌比较,经过包埋后的两歧双歧杆菌在模拟胃液和高胆盐溶液中的存活率分别提高了5个和2个对数值。     

Influence of ultrasound and enzymatic cross-linking on freeze-thaw stability and release properties of whey protein isolate hydrogel

This study investigated the effect of ultrasound and enzymatic cross-linking on the freeze-thaw (FT) stability and release properties of whey protein isolate hydrogels. We evaluated the FT stability by the changes in the microstructure, riboflavin retention, syneresis, water holding capacity (WHC), and texture of gels subjected to 3 FT cycles. High-intensity ultrasound (HUS) and transglutaminase (TGase)-mediated cross-linking improved the FT stability of whey protein isolate hydrogels loaded with riboflavin (WPISAR), as demonstrated by a more uniform and denser porous structure, significantly higher riboflavin retention, WHC, and textural properties, and lower syneresis after 3 FT cycles than those of untreated hydrogels. Furthermore, HUS- and TGase-mediated cross-linking decreased protein erosion and swelling ratio of WPISAR in simulated gastrointestinal fluids (SGIF) and reduced the riboflavin release rate in SGIF both with and without the addition of digestive enzymes. After 3 FT cycles, faster riboflavin release occurred due to a more porous structure induced by ice crystal formation compared with their unfrozen counterparts as detected by confocal laser scanning microscopy. High-intensity ultrasound- and TGase-mediated cross-linking alleviated the FT-induced faster riboflavin release rate in SGIF. High-intensity ultrasound- and TGase-treated gel samples showed that both diffusion and network erosion were responsible for riboflavin release regardless of FT. These results suggest that HUS- and TGase-mediated cross-linking improved the FT stability of WPISAR with a high riboflavin retention, and might be a good candidate as a controlled-release vehicle for riboflavin delivery to overcome undesired FT processing.