Physical and Molecular Properties of Egg-white Lipid Films

Polyethylene glycol-plasticized films were cast from alkaline (pH 11.25), heated (45 °C for 20 min), aqueous egg white (EW) solutions, with or without (10% or 20% w/w of EW) milkfat (two fractions), oleic acid, or egg yolk lysophospho-lipid (LPL). The lipids did not reduce (P > 0.05) film water vapor permeability. Oleic acid increased (P < 0.05) tensile strength and elongation, and surface sulfhydryl group (SH) concentrations in EW solutions. Oleic acid probably increased negative charges on EW proteins, unfolding protein chains, and exposing SH groups. LPL also slightly increased (P < 0.05) surface SH concentrations in non-heated mixtures. Electrophoretic patterns suggested oleic acid interactions with ovalbumin, ovotransferrin, and lysozyme. No lysinoalanine was in film-forming mixtures based on lysine measurements.     

Physical and Molecular Properties of Wheat Gluten Films Cast from Heated Film-Forming Solutions

The effect of heating film-forming solutions on physical and molecular properties of cast wheat gluten (WG) films was determined. Glycerol-plasticized films were cast from alkaline (pH 10), heat-treated (55,75, or 95°C for 10 min) solutions of WG in aqueous ethanol. Protein solubility (PS) of films in water decreased (P<0.05) with increasing temperature. Gel permeation chromatograms showed reduced extractability of protein fractions other than ω-gliadins in WG films. This reduced extractability was due to disulfide (S-S) bond formation. SDS-PAGE patterns of native WG and WG film samples suggested increased cross-linking through covalent S-S bonds in films from solutions heated at 75 or 95°C. Water resistance in potential packaging applications of WG edible films could be modified by adjusting heat-treating temperature.     

Frozen Stability of Fish Protein Isolate Under Various Storage Conditions

ABSTRACT: A novel process using pH-shift to recover fish proteins has been intensively studied. However, little information of its frozen stability has been revealed. The highest gel texture was found for samples frozen at pH 5.5 (5) and 7.0 (7) with cryoprotectants (C) and without freeze/thaw (F), whereas the lowest gel texture was obtained from frozen/thawed samples without cryoprotectants (NC). 5NC-F and 7NC-F demonstrated the lowest surface hydrophobicity and total sulfhydryl (SH), perhaps suggesting more protein aggregation through hydrophobic interactions and disulfide bonds. A slightly less stability of alkali-treated protein isolates (AKPI) kept at pH 5.5 than at pH 7 was noticed. AKPI, whether kept at pH 5.5 or 7.0, requires cryoprotectants to maintain frozen stability for longer shelf life.     

提取条件对大豆11S 球蛋白得率及纯度的影响

以低脂质含量豆粕为原料,探讨大豆11S球蛋白分级分离过程中提取条件(pH值、还原剂和冷沉时间)对蛋白得率及纯度的影响。结果表明：将浸提液pH值由6.2调至6.6时,大豆11s球蛋白的得率降低了16.71%,而纯度提高了4.60%。随着浸提pH值的提高,蛋白质的巯基与二硫键含量增加。与二硫苏糖醇(DTT)或β-巯基乙醇(ME)相比,在pH6.4的浸提液中添加亚硫酸氢钠(SBS),大豆11s球蛋白的得率最高。而DTT、ME和SBS还原剂中,使用DTT时的11S球蛋白组分得率与纯度最低。在蛋白沉淀阶段,当冷沉时间为10h时,蛋白的得率与纯度最佳,随着冷沉时间的延长(12～16h)巯基与二硫键总量并无显著差异(P≥0.05),而部分巯基发生氧化形成二硫键,大豆11S球蛋白游离巯基含量降低。     

Effect of Various Types of Egg White on Characteristics and Gelation of Fish Myofibrillar Proteins

ABSTRACT:  Three types of egg white protein (regular dried egg white [REW], special dried egg white [SEW], and liquid egg white [LEW]) were compared for their effect in surimi (CON), containing no egg white (EW). They were characterized for enzyme inhibition and time of EW addition, either with cryoprotectants prior to freezing or during gel preparation, using Pacific whiting surimi. In addition, the setting (suwari) effect and fish protein–EW protein interactions (dynamic rheological properties, total sulfhydryl groups, and fracture gel analysis) were evaluated using Alaska pollock surimi. After 12 mo frozen storage, adding 2% and 3% SEW to Pacific whiting surimi during chopping significantly ( P  < 0.05) increased the force and deformation values compared to adding the respective EW before freezing. The highest ( P  < 0.05) force (175.2 g) and deformation (9.0 mm) values after 12 mo were obtained when 3% SEW was added during chopping. Enzyme autolysis showed that addition of 2% EW protein was effective at inhibiting enzyme activity. During setting, adding 2% SEW maintained force (1047.4 g) and deformation (17.9 mm) values similar ( P  > 0.05) to CON (1055.1 g; 19.3 mm) and significantly ( P  < 0.05) better than 2% REW (666.1 g; 15.6 mm). Texture results corresponded well to other data where 2% SEW showed the lowest total sulfhydryl groups (48.3 mole/10⁵ g), and higher elastic modulus (G'), which suggests improved protein interaction for gel formation. Overall, the addition of 2% to 3% SEW improved gel textural properties of Pacific whiting and Alaska pollock fish protein.     

Thermal Aggregation of Soybean (Glycine max L.) Sulfur-rich Protein

The sulfur-rich protein (SRP) in soybeans aggregated at ≥ 70°C due to thermal denaturation of the protein and subsequent formation of soluble and insoluble aggregates. Aggregation was dependent on heating temperature, time, buffer composition, and the presence or absence of protein modifying agents in the heating buffer. Hydrophobic interactions and disulfide bonds helped stabilize the protein against thermal aggregation while hydrogen bonds and sulfhydryl exchange promoted it. At pH 7.6, 4.5 mg protein/mL, 70°C and 1.0M salt concentration, the rate of thermal aggregation was dependent on the anion type and followed lyotropic series SCN-> I-> Br-> Cl^?     

Gel-Forming Characteristics of Milk Proteins, 2. Roles of Sulfhydryl Groups and Disulfide Bonds

The gel-forming characteristics of milk proteins were investigated by employing skim milk either nonpreheated or preheated at 80°C, and coagulating them at 70 or 80°C with glucono-delta-lactone. The solubility of each gel in the phosphate buffer (pH 7.0) containing urea and 2- mercaptoethanol was examined. The disulfide bonds played a more important role in the gel coagulated at 80°C from skim milk preheated at 80°C than in the gel coagulated at 70°C from nonpreheated skim milk.The effect of the reduction treatment with 2-mercaptoethanol was more pronounced on preheated skim milk than on nonpreheated skim milk. Sulfhydryl groups and disulfide bonds, which were buried in the molecules of whey proteins in their native state, were rendered accessible following heat treatment at 80°C.The gel prepared from skim milk pretreated with oxidizing agent, hydrogen peroxide (i.e., skim milk has no accessible sulfhydryl groups as a result), or the gel prepared from skim milk pretreated with reducing agent, 2-mercaptoethanol, (i.e., skim milk has few disulfide bonds as a result), displayed weak gel formation. But the gel prepared from the mixture of these skim milks with appropriate ratio displayed higher gel firmness. These findings suggest that intermolecular disulfide bonds are formed by the exchange reaction between sulfhydryl groups and disulfide bonds during gel formation.     

茶多酚改性胶原蛋白-壳聚糖复合膜对冷藏斜带石斑鱼的保鲜效果

研究茶多酚改性后的胶原蛋白-壳聚糖复合膜对冷藏斜带石斑鱼的保鲜效果。在4℃冷藏条件下,采用空白对照、胶原蛋白-壳聚糖复合膜、茶多酚改性胶原蛋白-壳聚糖复合膜3种方式处理样品并贮藏20 d,以K值、肌动球蛋白含量、巯基(—SH)含量、Ca~(2+)-ATPase活性以及p H值、硫代巴比妥酸(thiobarbityric acid,TBA)值、挥发性盐基氮(total volatile basic nitrogen,TVB-N)值、菌落总数、感官评定值作为研究指标。结果表明:胶原蛋白-壳聚糖复合膜、茶多酚改性胶原蛋白-壳聚糖复合膜处理组斜带石斑鱼的巯基含量、肌动球蛋白含量、Ca~(2+)-ATPase活性和感官评定值均显著高于对照组(P0.05),而K值、p H值、TVB-N值、菌落总数均显著低于对照组(P0.05),且茶多酚改性胶原蛋白-壳聚糖复合膜处理组的效果优于胶原蛋白-壳聚糖复合膜处理组,其中3个处理组的TBA值的差异性不显著(P0.05)。茶多酚改性胶原蛋白-壳聚糖复合膜处理组鱼肉的肌动球蛋白变性和巯基含量降低缓慢、Ca~(2+)-ATPase活性下降受到阻碍、TVB-N值等理化指标变化以及细菌总数生长受到抑制。说明在4℃冷藏条件下,茶多酚改性胶原蛋白-壳聚糖复合膜处理组能有效延缓斜带石斑鱼的腐败变质,保鲜效果更明显。     

豌豆蛋白对猪肉盐溶蛋白理化性质的影响

以猪背最长肌为原料提取盐溶蛋白,研究添加2%、4%、6%、8%、10%的豌豆蛋白对猪肉盐溶蛋白的SDS-PAGE、溶解度、表面疏水性、巯基、二硫键含量以及乳化性等的影响。结果表明,随豌豆蛋白添加量的增加,盐溶蛋白中肌球蛋白重链、C-蛋白、α-肌动素、肌球蛋白轻链条带逐渐变窄且颜色变浅;盐溶蛋白中的肌动蛋白与豌豆球蛋白(vicilin 7S)亚基,盐溶蛋白中的肌钙蛋白T与豌豆蛋白中酸性多肽(legA),分子量相近,其条带逐渐变粗且颜色加深。与未添加豌豆蛋白的盐溶蛋白相比,当豌豆蛋白添加量为2%~6%时,盐溶蛋白的表面疏水性和乳化性显著上升(p<0.05),豌豆蛋白添加量为8%~10%时,差异不显著(p>0.05);随豌豆蛋白添加量的增加,浊度、二硫键含量均呈上升趋势,且二硫键在添加量为10%时最高,为2.196 mol/105 g;总巯基和自由巯基含量下降;溶解度则先升高后下降,在豌豆蛋白添加量为4%时达到最大值,为42.88%。可见,添加适量的豌豆蛋白对猪肉盐溶蛋白的理化性质有显著影响。     

脂质过氧自由基诱导兔肉肌浆蛋白聚集机制

为全面理解兔肉品质的形成过程,利用偶氮二异丁脒盐酸盐（2,2’-azobis(2-amidinopropane)dihydrochloride,AAPH）的热分解作用生成过氧自由基（ROO·）诱导兔肉肌浆蛋白氧化,通过AAPH处理后兔肉肌浆蛋白中高铁肌红蛋白相对含量、超铁肌红蛋白浓度、羰基含量、总巯基含量以及肌浆蛋白表面疏水性、粒度、浊度和溶解性测定,肌浆蛋白十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE）和内源荧光光谱分析,研究ROO·胁迫肌浆蛋白氧化聚集机制。结果表明,随着AAPH浓度的升高,高铁肌红蛋白和超铁肌红蛋白相对含量显著增加（P＜0.05）,蛋白质羰基含量显著增加（P＜0.05）,总巯基含量显著下降（P＜0.05）。内源荧光光谱和表面疏水性分析结果表明ROO·可以引起肌浆蛋白的去折叠和疏水基团暴露。SDS-PAGE分析结果表明ROO·可以导致肌浆蛋白二硫键交联。此外,随着AAPH浓度的增大,肌浆蛋白粒度和浊度呈现出逐渐增大的趋势,表明ROO·可以引发肌浆蛋白的聚集。疏水相互作用和二硫键交联是ROO·胁迫肌浆蛋白聚集的主要作用力。ROO·不仅可以直接引发肌浆蛋白聚集,还可以通过影响肌红蛋白诱导氧化体系促进肌浆蛋白交联聚集。     

Effect of solution pH on solubility and some structural properties of soybean protein isolate films

Changes in solubility and molecular properties of protein films obtained from soy protein isolate (SPI) solutions at different pH values (2, 8 and 11) were investigated to study protein behavior during film formation. Proteins retained their native conformation in films at pH 8, but were partially or extensively denatured at pH 11 and 2. Although film protein networks were maintained by the same type of interactions at different pH values—covalent (disulfide bonds) and non‐covalent bonds (especially hydrophobic interactions and hydrogen bonds)—the intensity of each type of interaction (predicted from solubility tests in buffers with different chemical action) depended on the pH of the initial solution. Films obtained at pH 8 presented the highest solubility in all the buffers, whereas films at formed pH 2 presented the lowest, except in the buffer of pH 8 that contained urea, SDS and 2‐mercaptoethanol, which totally dissolved 100% of the film proteins. Films prepared at extreme pH values had a denser microstructure than those at pH 8. SDS–PAGE patterns indicated that films were mainly formed by β‐conglycinin and glycinin, which aggregated in different forms during film formation, depending on the pH of the initial solutions. Some of these proteins remained weakly bonded and/or were held by the network of films. These differences in the protein networks structure would affect the physical, mechanical and barrier properties of the films. Copyright © 2006 Society of Chemical Industry     

离子强度对猪肉肌原纤维蛋白乳化特性和理化特性的影响

以猪肉肌原纤维蛋白为研究对象,测定不同离子强度下肌原纤维蛋白的乳化特性,包括乳化能力（emulsifying capacity,EC）、乳化稳定性（emulsifying stability,ES）,同时测定不同离子强度下肌原纤维蛋白的溶解性、分子间氢键、表面疏水性、活性巯基和总巯基等理化特性,并对理化特性、乳化特性指标进行了相关性分析。结果表明：随着介质离子强度的提高,肌原纤维蛋白的EC和ES增强,溶解度增大,分子间氢键、活性巯基呈上升趋势,表面疏水性则呈下降趋势,而总巯基则无明显变化;溶解度与EC显著正相关（P＜0.05）,与ES呈极显著正相关（P＜0.01）,活性巯基和氢键都与EC呈极显著正相关（P＜0.01）,而表面疏水性与EC呈极显著负相关（P＜0.01）,与ES显著负相关（P＜0.05）。结论：改变肌原纤维蛋白介质的离子强度,导致肌原纤维蛋白的乳化特性、理化特性产生变化,肌原纤维蛋白的理化特性和乳化特性之间有显著或极显著的相关性。     

超高压结合热处理对猪肉蛋白质相互作用力及结构的影响

以猪背最长肌为原料,研究超高压(100~600 MPa)结合热处理(25~55℃)条件下,蛋白质相互作用力的变化,猪肉主要组成部分肌原纤维蛋白氧化程度,表面疏水性及结构变化,并建立他们之间的相关性。研究结果表明:超高压结合热处理后稳定猪肉蛋白质的作用力主要是氢键、疏水性相互作用和二硫键。经超高压结合热处理的肌原纤维蛋白质巯基含量极显著下降(p0.01),二硫键含量、疏水性和羰基值极显著上升(p0.01)。二级结构发生重排,螺旋结构减少,无序结构增多。相关性分析结果表明超高压结合热处理与蛋白质相互作用力、蛋白质羰基值、蛋白质结构之间存在着极显著的相关关系(p0.01)。这些结果可以帮助我们全面认识超高压结合热处理过程中蛋白质的变化,科学指导其生产过程,为开发新型肉制品提供理论依据。     

氧化强度对肌原纤维蛋白结构及凝胶性能的影响

采用模拟氧化体系（10 μmol/L FeCl3,100 μmol/L抗坏血酸,0、0.5、1、3、5、10 mmol/L H2O2）,研究不同氧化程度对猪肌原纤维蛋白（myofibrillar protein,MP）结构及凝胶性质的影响。结果表明：随H2O2浓度升高,蛋白羰基含量上升,总巯基含量、自由氨基含量及内源荧光强度下降,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表明氧化诱导二硫键的形成引起蛋白质交联聚集加剧,进而导致蛋白表面疏水性和溶解度降低;当H2O2浓度小于1.0 mmol/L时,MP热诱导凝胶性能无明显变化,当H2O2浓度大于1.0 mmol/L时,热诱导凝胶蒸煮损失显著增强（P＜0.05）,凝胶强度明显降低（P＜0.05）,但凝胶白度无明显变化。总体来说,H2O2浓度越高蛋白氧化程度越严重,凝胶强度越低。     

Effect of High Pressure and/or Temperature over Gelation of Isolated Hake Myofibrils

High hydrostatic pressure (HHP) processing was used to determine its ability to induce protein gelation. Isolated hake myofibrils were processed by HHP at 0, 150, 250, and 500 MPa (10?ºC/10 min) and/or by heating (90?ºC/20 min). The results were analyzed by Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), determination of sulfhydryl group contents, and dynamic rheometry measurements. FTIR data indicated that secondary protein structures exhibited a reduction in α-helix together with an increase in β-sheet as a result of protein denaturation caused by HHP. DSC showed that HHP induced a reduction in myosin denaturation temperature (T _peak) indicating protein unfolding. Protein gelation after HHP is based on physical (non-covalent) interactions which make more sulfhydryl groups available, while after heating, it is based on the formation of covalent (disulfide) bonds as a consequence of protein denaturation reducing the sulfhydryl groups. The combination of HHP and heating, particularly the latter, improved network stabilization. These results were reflected in the rheological changes, in which heated gels showed more elastic, cohesive, and time-stable networks than pressurized (non-heated) gels. The HHP effect provided softer, more flexible networks. The gel at 500 MPa was the most elastic and time-stable and exhibited the highest level of connectivity.     

超声波和TGase对重组草鱼肉形成过程中理化性质的影响

本研究以草鱼为原料,采用四种处理方式:对照组(只添加NaCl)、实验组1(添加NaCl和谷氨酰胺转氨酶(transglutaminase,TGase))、实验组2(添加NaCl且超声)、实验组3(添加NaCl和TGase且超声),对草鱼肉进行重组。通过测定其分子间作用力、巯基含量、浊度、溶解率等,分析重组过程中超声波和TGase对重组草鱼肉理化性质的影响。结果表明,在重组草鱼肉形成过程中,超声波和TGase对离子键、氢键、疏水相互作用、二硫键和非二硫键均有不同程度的影响,促进重组草鱼肉的形成;与此同时,在重组草鱼肉形成过程中,超声波和TGase可以加速肌原纤维蛋白巯基氧化形成二硫键,增强其表面疏水性,增加其浊度,降低重组草鱼肉蛋白质溶解率;由SDS-PAGE电泳图谱可知,在重组草鱼肉形成过程中,三个实验组在200 kDa附近的条带逐渐变淡,甚至消失,可知重组过程中超声波和TGase主要作用于肌原纤维蛋白重链。综合各试验指标结果,超声波和TGase均可促进草鱼肉蛋白质交联,且两者同时作用时交联度最高,对指导草鱼肉产业生产具有一定的参考价值。     

Correlation of Protein Sulfhydryls with the Strength of Heat-Formed Egg White Gels

A strong, direct correlation (r = 0.97) was found between egg white protein surface sulfhydryl group (-SH) concentration and the gel strength of its heat-induced gels. An inverse correlation (r = - 0.86) was also found between the -SH group concentration of 0.1% SDS-denatured egg white proteins and gel strength. Data indicated that surface - SH groups played an important role in strong heat-induced gel formation in egg whites.     

脱酰胺处理对不同谷物蛋白体外消化率的影响

脱酰胺处理是蛋白质改性一种方式,经脱酰胺后,蛋白的理化性质和营养性质都会发生改变。本实验研究了酸法脱酰胺处理对大米、小麦、玉米蛋白体外消化率的影响,并从蛋白质的溶解度、表面疏水性、巯基和二硫键含量以及超微结构等理化性质方面来探究脱酰胺影响体外消化率的作用机理。试验结果表明:随着脱酰胺程度的加大,大米蛋白、小麦蛋白和玉米蛋白质巯基和二硫键之间的相互转化不明显(P0.05),两者的含量基本维持不变;三种谷物蛋白的溶解度在胃蛋白酶最适p H 1.5和胰蛋白酶最适p H 7的条件下逐渐增大;蛋白质的表面疏水性随脱酰胺度的增加逐渐减小;体外消化率也分别从原来的66.5%、79.5%和78.5%提高到了79.5%、89.0%、和89.2%,超微结构从原来的致密聚集变得多孔疏松,更利于蛋白质的消化。     

杏鲍菇分离蛋白和清蛋白的理化性质及功能分析

以杏鲍菇为原料,分别利用碱溶酸沉法提取杏鲍菇分离蛋白（Pleurotus eryngii protein isolate,PEPI）、Osborne法分离主要蛋白组分,并研究其理化性质和功能分析。结果表明,杏鲍菇的蛋白质量分数为17.57%（以干质量计）,以杏鲍菇清蛋白（Pleurotus eryngii albumin,PEA）为主,占总分离蛋白组分的81.12%。PEPI和PEA中均含18?种氨基酸,且必需氨基酸含量分别占总氨基酸含量的40.80%和40.51%。与PEPI相比,PEA的表面疏水性（265.25）显著高于PEPI（164.27）（P＜0.05）,而总巯基、二硫键含量较低,分别为61.53?μmol/g和10.39?μmol/g;热变性温度（100.98?℃）低于PEPI（108.27?℃）,且PEA持水性（1.64?mL/g）、持油性（5.59?mL/g）显著低于PEPI（3.58、8.36?mL/g）（P＜0.05）。PEPI和PEA的溶解性、起泡性、泡沫稳定性、乳化性及乳化稳定性随pH值的变化趋势均相似,在等电点时均为最低。傅里叶变换红外光谱显示PEPI和PEA的二级结构主要是β-折叠和β-转角,扫描电镜观察PEPI呈蜂巢结构。相比PEA,PEPI具有更好的理化性质和功能特性。     

低温冷冻条件对大豆分离蛋白分散液表面疏水性及二硫键的影响

通过大豆分离蛋白（soybean protein isolate,SPI）冷冻前后结构的变化,研究了低温冷冻条件（SPI添加量、冷冻温度、冷冻时间）对SPI结构（巯基、二硫键及表面疏水性）的影响。实验发现在冷冻条件下随着SPI添加量的降低,游离巯基和二硫键逐渐减少、暴露巯基与表面疏水性先增加后减小;随着冷冻温度降低和冷冻时间延长,SPI的游离巯基、二硫键含量及表面疏水性逐渐减少。经过冷冻处理的SPI会有一定程度的变性,在不太低的冷冻温度下游离巯基含量明显大于未冷冻SPI,且在冷冻过程中SPI大部分的分子内二硫键转化为分子间二硫键。