Effect of acetylation and succinylation on solubility profile, water absorption capacity, oil absorption capacity and emulsifying properties of mucuna bean (Mucuna pruriens) protein concentrate

Mucuna protein concentrate was acylated with succinic and acetic anhydride. The effects of acylation on solubility, water absorption capacity, oil absorption capacity and emulsifying properties were investigated. The pH-dependent solubility profile of unmodified mucuna protein concentrate (U-mpc) showed a decrease in solubility with decrease in pH and resolubilisation at pH values acidic to isoelectric pH (pH 4). Apart from pH 2, both acetylated mucuna protein concentrates (A-mpc) and succinylated mucuna protein concentrate (S-mpc) had improved solubility over the unmodified derivative. Acylation increased the water absorption capacity (WAC) at all levels of ionic strength (0.1-1.0 M). WAC of the protein samples increased with increase in ionic strength up to 0.2 M after which a decline occurred with increase in ionic strength from 0.4-1.0 M. When protein solutions were prepared in salts of various ions, increase in WAC followed the Hofmeister series in the order: NaSCN < NaClO4 < NaI < NaBr < NaCl < Na2SO. Acetylation improved the oil absorption capacity while the lipophilic tendency reduced the following succinylation. Emulsifying capacity increased with increase in concentration up to 2, 4 and 5% w/v for U-mpc, A-mpc and S-mpc, respectively, after which an increase in concentration reduced the emulsifying capacity. Both acetylation and succinylation significantly (P < 0.05) improved the emulsifying capacity at pH 4-10. Initial increase in ionic strength up to 0.4 M for U-mpc and 0.4 M for A-mpc and S-mpc increased the emulsion capacity progressively. Further increase in ionic strength reduced emulsion capacity (EC). Contrary to the effect of various salts on WAC, increase in EC generally follows the series Na2SO4 < NaCl < NaBr < NaI < NaClO4 < NaSCN. At all levels of ionic strength studied, S-mpc had a better emulsifying activity (EA) than both A-mpc and U-mpc. EA and emulsifying stability (ES) were pH-dependent. Maximum EA and ES were recorded at pH 10. ES of protein derivatives were higher than those of U-mpc in the range of pH 4-10 but lower at pH 2. Studies revealed that both A-mpc and S-mpc had better ES and EA than the unmodified derivative when protein solutions were prepared in salts of various anions.     

Lipophilization of β-lactoglobulin: Effect on allergenicity and digestibility

β-Lactoglobulin was chemically modified by covalent attachment of different levels of stearic acid and the effect on allergenicity and digestibility of the protein was evaluated. A decreased in vitro digestibility was observed with extent of lipophilization (stearic acid incorporation). Increased ability to elicit immunoglobulin E (IgE) antibodies, determined by heterologous passive cutaneous anaphylaxis, was observed at low lipophilization (attachment of fatty acid). Medium level of lipophilization decreased this ability and at high lipophilization the ability to elicit IgE antibodies and was almost destroyed. Low and medium lipophilization increased immunoglobulin G (IgG) binding ability as measured by enzyme-linked immunosorbent assay. High lipophilization decreased the IgG binding ability.     

The acylated protein derivatives of Canavalia ensiformis (jack bean): A study of functional characteristics

Protein concentrate was prepared from jack bean (JNP) and it was modified by acylation using acetic (JAP) and succinic anhydrides (JSP). Proximate analyses revealed that moisture and ash content increased following acetylation and succinylation, while both acetylation and succinylation reduced percentage crude fat and protein. Acetylation and succinylation reduced protein solubility in the acidic pH range below the isoelectric point (4.5) of the protein concentrate but improved the solubility of the unmodified protein concentrate at the isoelectric point and pH range alkaline to the isoelectric point. Both acetylation and succinylation increased the water absorption capacity of unmodified protein concentrates at all levels of ionic strength investigated (0.1-1.0 mol/l). Acetylation improved oil absorption capacity whereas the tendency to absorb oil reduced after succinylation. Maximal emulsifying activity of native and modified proteins were obtained at pH 10. Emulsion stability of acylated proteins was higher than those of native proteins in the range of pH 4-10 but lower when the pH was 2. Foam capacity and stability of both native and modified proteins increased with increase in protein concentration. Foam capacity of modified proteins increased progressively with increase in pH from 2 to 10. Also, acylated protein derivatives had improved foam capacity over the native protein except at pH 2. Gelation capacity of both native and modified proteins was maximal at the region of isoelectric point.     

Effect of succinylation on the functional and physicochemical properties of alpha-globulin,the major protein fraction from Sesamum indicum L

alpha-Globulin the major protein fraction from Sesamum indicum was succinylated to different levels and the effect of the chemical modification was evaluated both on the functional and physicochemical properties. The results suggest that the pH of minimum solubility shifted to the more acidic side (pH approximately 4.5-5.5) for the succinylated alpha-globulin whereas for control alpha-globulin the pH of minimum solubility was 6.5. Succinylation also increased emulsion activity and emulsion stability of the protein. The emulsion stability increased from a control value of 53 +/- 3 s to a value of 122 +/- 5 s. Bulk density, water absorption capacity, oil absorption capacity, foam capacity and foam stability were evaluated in phosphate buffer (pH 7.0) containing 0.5 M sodium chloride and all these properties showed increased values as a result of succinylation. Ultracentrifugation studies showed that the % composition of 7S component increases with concomitant decrease in that of 11S fraction with the increase in percentage of succinylation. Further increase in succinylation resulted in only 2S component which is a dissociated form of 11S and/or 7S protein fractions. The fluorescence emission studies showed a decrease in the fluorescence emission intensity of alpha-globulin as a result of succinylation. The thermal stability of the protein molecule decreased due to progressive succinylation as indicated by decrease in the apparent thermal denaturation temperature from a control value of 84 to 62 degrees C at a succinylation level of 40%. These results suggest that succinylation improves the functional characteristics of alpha-globulin. Such changes in the functional properties have been attributed partly to the dissociation of the protein molecule at higher levels of succinylation and the increase in the net negative charge on the protein.     

Effect of succinylation on the functional and physicochemical properties of α‐globulin,the major protein fraction from Sesamum indicum L.

α‐Globulin the major protein fraction fromSesamum indicum was succinylated to different levels and the effect of the chemical modification was evaluated both on the functional and physicochemical properties. The results suggest that the pH of minimum solubility shifted to the more acidic side (pH ˜ 4.5–5.5) for the succinylated α‐globulin whereas for control α‐globulin the pH of minimum solubility was 6.5. Succinylation also increased emulsion activity and emulsion stability of the protein. The emulsion stability increased from a control value of 53 ± 3 s to a value of 122 ± 5 s. Bulk density, water absorption capacity, oil absorption capacity, foam capacity and foam stability were evaluated in phosphate buffer (pH 7.0) containing 0.5 M sodium chloride and all these properties showed increased values as a result of succinylation. Ultracentrifugation studies showed that the % composition of 7S component increases with concomitant decrease in that of 11S fraction with the increase in percentage of succinylation. Further increase in succinylation resulted in only 2S component which is a dissociated form of 11S and/or 7S protein fractions. The fluorescence emission studies showed a decrease in the fluorescence emission intensity of α‐globulin as a result of succinylation. The thermal stability of the protein molecule decreased due to progressive succinylation as indicated by decrease in the apparent thermal denaturation temperature from a control value of 84 to 62°C at a succinylation level of 40%. These results suggest that succinylation improves the functional characteristics of α‐globulin. Such changes in the functional properties have been attributed partly to the dissociation of the protein molecule at higher levels of succinylation and the increase in the net negative charge on the protein.     

木糖醇对大豆分离蛋白结构和起泡特性的影响

研究添加不同质量分数木糖醇时大豆分离蛋白的结构、溶解性、表面疏水性、内源荧光强度以及起泡特性的变化情况,以期更加深入了解木糖醇对大豆分离蛋白结构和功能特性的影响。结果表明：在木糖醇的作用下,大豆分离蛋白的溶解性增加,而表面疏水性和内源荧光强度降低,原来暴露的酪氨酸和色氨酸残基则被包裹到分子内部。同时,大豆分离蛋白的结构也发生了改变,其二级结构变得更加有序、致密。由于大豆分离蛋白结构的变化,其起泡能力受到抑制。另外,木糖醇使大豆分离蛋白溶液的表观黏度增加,有利于提高其泡沫稳定性。     

海鲜菇蛋白质功能特性及消化性的研究

以海鲜菇粉为原料,采用纤维素酶辅助碱提酸沉法提取出蛋白质,对其功能特性及消化性进行研究。结果表明,随着离子强度的增加,海鲜菇蛋白质的溶解性(Solubility)、持水性(Water holding)、乳化性(Emulsifying)及乳化稳定性(Emulsion stability)、起泡性(Foaming)及泡沫稳定性(Foaming stability)均呈现先升高后降低的趋势;随着蔗糖浓度的提高,溶解性逐渐降低,持水性、乳化性、起泡性及泡沫稳定性呈现先升高后降低的趋势,乳化稳定性呈现先降低后升高的趋势;随着温度的提高,溶解性、持水性、持油性(Oil holding)、乳化性及乳化稳定性、起泡性及泡沫稳定性均呈现先升高后降低的趋势;随着pH的升高,溶解性、起泡性、乳化性及乳化稳定性均呈现先降低后升高的趋势,在等电点附近达到最小值,泡沫稳定性呈现先升高后降低的趋势,在等电点附近达到最大值。海鲜菇蛋白质溶液经过模拟胃消化后,其消化率为35.5%,水解度17.13%,游离氨基酸含量0.42 mg/mL,多肽含量1.16 mg/mL,胃消化液还原力0.55,DPPH·、O2-·和·OH的清除率分别为66.14%、67.36%和69.44%;模拟肠消化后,其消化率为87.65%,水解度20.88%,游离氨基酸含量0.93 mg/mL,多肽含量0.83 mg/mL,肠消化液的还原力0.23,DPPH·、O2-·和·OH的清除率分别为35.02%、27.17%和37.75%。通过试验证明了离子强度、温度、时间和蔗糖浓度对海鲜菇蛋白质的功能特性有一定影响;该蛋白质在胃肠中消化率较高,是一种易消化优质蛋白质。     

Functional and electrophoretic characteristics of faba bean (Vicia faba) flour proteins as affected by germination

Faba beans (Vicia faba) were germinated at room temperature for 3 and 6 days respectively. The effect of germination on the protein fractions, protein solubility index, PAGE pattern and some functional properties i.e. emulsification capacity (EC), foaming capacity (FC), foam stability (FS), water and fat absorption capacities of the flour was studied. Germination decreased albumins, globulins and prolamins at different levels but non protein nitrogen and glutelins were increased. The protein solubility index was high at both extreme pH values with an isoelectric point (IP) at pH of 4.4-4.5. The solubility of the protein slightly increased due to germination at all the pH values. PAGE pattern revealed on obvious dissociation and utilization for both fast and slow moving protein fractions during germination. Emulsification and foaming properties vs pH profile were similar to the pattern of solubility vs pH. Both properties were high at acidic and alkaline pH's and the minimum values were at pH 4 to 5. Germination process improved EC, FC and FS of the flour in comparison with that of dry bean flour. Water absorption of faba bean flours was improved during germination but the fat absorption markedly decreased.     

Effect of Partial Proteolysis and Succinylation on Functionality of Corn Germ Protein Isolate

Corn germ protein isolate (CGPI) was partially hydrolyzed with trypsin and pepsin and succinylated at three levels. Various functional and electrophoretic properties of the native and modified protein were determined. Water absorption and foaming properties of CGPI were Improved by partial hydrolysis with trypsin; emulsifying capacity and nitrogen solubility were reduced; oil absorption was increased only slightly. CGPI pepsin hydrolyzate has decreased oil absorption, nitrogen solubility and emulsifying capacity but improved foaming properties; water absorption was unchanged. Treatment of CGPI with succinic anhydride improved water and oil absorption, nitrogen solubility and foaming capacity but decreased emulsifying capacity; foam stability was unchanged. Succinylation retarded electropohoretic mobility while hydrolysis altered band intensities.     

中国毛虾酶解产物的功能性质研究

研究了木瓜蛋白酶酶解中国毛虾产物的功能特性。对毛虾酶解产物的溶解性、乳化性、乳化稳定性、起泡性、泡沫稳定性和粘度等功能性质进行了研究,并与虾粉的功能特性进行了对比。结果表明:毛虾酶解产物的蛋白质含量高达85.5%,比虾粉提高17.5%;酶解产物的溶解度、乳化性和起泡性明显高于虾粉;酶解产物的乳化稳定性、泡沫稳定性和粘度略低于虾粉。毛虾酶解产物作为一种潜在的功能性配料,在食品工业中具有一定的应用前景。     

Functionality of phosphorylated vicilin exposed to chemical and physical agents

The effects of phosphorylation with sodium trimetaphosphate (STMP) on functional and physicochemical properties of pea vicilin, as probed by high hydrostatic pressure and chemical denaturation were evaluated. The isoelectric point of unmodified and phosphorylated vicilin decreased in the presence of 0.5 M NaCl, resulting in a decrease of the solubility at pH 1.0. The gelation capacity of unmodified vicilin in the presence of NaCl decreased approximately 80% when compared with unmodified vicilin without NaCl. Increasing pressure from 0.1 MPa (atmospheric pressure) to 240 MPa significantly decreased the solubility of vicilin phosphorylated with 4% STMP at pH 1.0 and 4.0 by about 30%. Pressure had no effect on solubility of native vicilin. Pressure treatment at 240 MPa improved the gelation capacity of vicilin phosphorylated with 1% STMP. Glycerol decreased the gelation capacity of vicilin and its solubility in the acidic pH range.     

Effects of succinylation and deamidation on functional properties of oat protein isolate

The effects of two different modification methods (deamidation and succinylation) on the functional properties (solubility, water- and oil-binding capacity, foaming capacity and stability, emulsion activity and stability) of oat protein isolates were evaluated. Protein isolates extracted from defatted oat flour at alkaline pH were acylated by 0.20 g/g of succinic anhydride. The protein isolate was also modified using a mild acidic treatment (HCl, 0.5 N). Succinylation and deamidation improved solubility and emulsifying activity of the native protein isolate. Foaming capacity of oat protein isolate increased after deamidation, whereas succinylation decreased it. The deamidated and succinylated proteins had lower foam and emulsion stabilities than had their native counterpart. Water- and oil-binding capacity, in both modified oat proteins, was higher than those of the native oat protein isolate.     

燕麦麸皮球蛋白的糖基化结构修饰及功能性变化

在氨基葡萄糖存在的条件下,利用转谷氨酰胺酶(transglutaminase,TG)对燕麦麸皮球蛋白糖基化结构修饰,进而探讨糖基化蛋白功能性质与结构之间的关系。结果表明,糖基化交联球蛋白的溶解性、乳化稳定性、起泡性及泡沫稳定性相比于未修饰的球蛋白都有明显的改善,但表面疏水性明显下降;另外,酶促糖基化球蛋白的变性温度和焓变值都有所下降,其二级结构变化为:α-螺旋结构相对含量呈增加趋势,β-折叠和β-转角结构相对含量呈下降趋势,无规卷曲结构相对含量几乎没变。经糖基化处理的球蛋白酪氨酸分子主要呈现"暴露态",色氨酸相对拉曼强度更趋近于"包埋态"。酶促糖基化球蛋白二硫键振动模式为t-g-t。通过对球蛋白、修饰球蛋白的功能特性与空间构象的比较分析,明确TG催化葡萄糖结合在燕麦麸球蛋白上,进一步明晰修饰蛋白功能特性与空间构象之间的构效关系。结果可为延长杂粮产业链提供良好的理论依据,同时可以为今后制备燕麦蛋白特定产品进行分子设计和重组提供基础数据。     

超高压处理对槟榔芋淀粉理化性质的影响

以槟榔芋淀粉为原料,采用超高压技术对淀粉进行改性处理,研究不同压力处理对其理化性质的影响.结果表明:随着压力的增大,槟榔芋淀粉的溶解度、膨胀度呈先减小后增大的趋势,但是均显著低于原淀粉;超高压处理可以显著增大槟榔芋淀粉的透光率;经200 MPa压力处理后,其冻融稳定性有明显改善.经300 MPa压力处理后,槟榔芋淀粉凝胶的硬度、咀嚼性和胶黏性都显著增加,但弹性和凝聚性变化不显著.RVA测定结果表明:淀粉糊的峰值黏度随处理压力的增大而显著增大;改性后槟榔芋淀粉的崩解值略高于原淀粉,而回生值变化不显著;200 MPa压力处理可降低槟榔芋淀粉的糊化温度.研究表明,一定程度的高压处理可以达到改善槟榔芋淀粉理化性质的目的.     

Functional properties of plant proteins. Part 2. Selected physicochemical properties of native and denatured protein isolates from faba beans,soybeans, and sunflower seed

Functional properties — as solubility, water and oil adsorption, emulsifying capacity, emulsion activity and stability — of protein isolates from faba beans, soybeans and sunflower seed depending on the isolation process were determined. Proteins isolated under mild conditions, it means by precipitation using dialysis or dilution of salt extracts with water, show the highest solubility, characterized by a sharp minimum of solubility at a rather narrow range of pH. An incubation of the precipitated proteins at low pH (pH 2) results in a decrease of the solubility on the alcaline and acidic part of the solubility profile. On the contrary to the decreased solubility, the proteins denatured by acid show an increased water adsorption capacity. Depending on the kind of protein and the conditions of preparation these values can reach the manifold ones of the control. Smaller increases of oil adsorption in acid-denatured proteins were found, too. The emulsion activity and stability were not or only slightly influenced, but the emulsifying capacity was strongly decreased by the denaturation procedure. The emulsifying capacity was influenced by the solubility of the protein, but a strong correlation does not exist. The high water adsorption of Promine D can be reached by the other plant proteins after denaturation. The sunflower protein showed the highest emulsifying capacity. Increasing the pH from the isoelectric range to 7 improves all studied functional properties.     

Effects of Acylation of Defatted Cottonseed Flour with Various Acid Anhydrides on Protein Extractability and Functional Properties of Resulting Protein Isolates

Maleylation, succinylation, dimethylglutarylation, and sodium sulfite treatment of cottonseed flour increased protein extraction from the flour in suspension, and precipitated more protein in the extract at pH 4.0, compared to unmodified flour processed through conventional methods. However, acetylation decreased protein extraction and precipitation at this pH. Protein isolates from succinylated, maleylated and dimethylglutarylated flours were highly water-soluble, and did not coagulate by heating. Acetylation decreased heat coagulability of the resulting protein isolate, but did not affect water solubility of the isolate. Treatment of the flour with sodium sulfite markedly increased heat coagulability of the isolate, and decreased its solubility. Oil absorption capacity of the isolate was increased slightly by dimethylglutarylation, but other treatments did not affect the capacity significantly. Sensitivity of proteins in isolates to calcium ions was not affected by acylation or sodium sulfite treatment of the flour.     

Enzymatic modification as a tool to improve the functional properties of heat‐processed soy flour

BACKGROUND: There are a number of antinutritional factors present in soybeans that exert a negative impact on the nutritional quality of the protein. Among those factors that are destroyed by heat treatment are protease inhibitors and lectins. Protease inhibitors show antinutritional effect and moreover the digestibility of the protein is limited by the presence of these antinutrients. The aims of the present study are (1) to study the effect of autoclaving on the trypsin inhibitor inactivation, nitrogen solubility and protein digestibility of defatted soy flour and (2) to study the effect of enzymatic modification on the functional properties of autoclaved soy flour. RESULTS: The solubility of the soy flour decreased with increase in autoclaving time. Partial hydrolysis of the autoclaved soy flour increased its acid solubility (pH 4.5) from 17% to 56% over a control value of 24% without affecting its functional properties. Inactivation of trypsin inhibitors improved the protein digestibility of soy flour from 25% to 95%. Particle size analysis of the autoclaved flour indicated the formation of soy protein aggregates, which resulted in poor solubility. The enzymatic modification of autoclaved soy flour resulted in its property as a good emulsifying agent with an emulsion capacity of 118 ± 4 mL. CONCLUSION: Enzymatic modification of the heat‐processed soy flour increased its solubility and other functional attributes. The increased acid solubility would be advantageous in the utilization of soy proteins in acidic foods. Thus the autoclaved and partially modified soy flour is a potential source for specific functional foods. Copyright © 2007 Society of Chemical Industry     

热诱导大豆蛋白纤维聚集体的分离及性质研究

酸性热处理条件下,随着大豆分离蛋白(SPI)自发形成纤维聚集体,仍有部分未自组装聚集的多肽分子存在其中,二者比例对蛋白功能性质产生较大影响。本文以超滤手段对热诱导大豆蛋白纤维聚集体和多肽进行大量分离,确定分离方法并研究两者的理化性质。结果表明:100 ku超滤膜反复分离两次即能获得较好的分离效果;SPI经热处理后,其等电点溶解度提高,但中性p H下溶解度变小,且蛋白水解致使其表面电位绝对值明显提高;分离所得纤维聚集体的氨基酸组成和表面电位与热处理蛋白相似,在等电点和中性p H溶解度更低;多肽则含有较少疏水氨基酸和较多负电氨基酸,在p H 2.0~9.0溶解度较好,其在酸性p H下电荷量和静电排斥力较低,导致其以无定形聚集体的形式存在,而中性p H其电荷量较高导致多肽分子间相互作用减弱,聚集体解离。     

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。     

Improving Foaming Properties of Yolk-Contaminated Egg Albumen by Basic Soy Protein

ABSTRACT:  Yolk contamination of egg white is a common problem in the egg breaking industry. Foaming properties of egg white protein are affected by such contamination, but proteins of basic nature may restore the foaming properties of the yolk-contaminated egg white protein. The purpose of this study was to chemically modify a soy protein, that is, to esterify the acidic groups on the protein and to study the potential of such modified protein in improving foaming. We showed that the modification changed the isoelectric point of soy protein isolate (SPI) from 4.5 to about 10. Sonication was proven to be a very effective means to redisperse the methanol-denatured soy protein during reaction, as shown by the improved solubility profile. Such modified basic protein, that is, the sonicated-modified SPI (SMSPI), when added to the yolk-contaminated (at 0.4% level, as-is basis) egg white, gave significantly improved foaming properties. We have shown that the slight change in pH due to the addition of SMSPI was not the reason for improved foaming performance; instead, the modified protein itself was the main reason for such improvement. Addition of SMSPI increased the foaming performance of both pure egg white and yolk-contaminated egg white. SMSPI consistently performed better than the unmodified SPI for improving foaming. Addition of SMSPI (16%, based on dry egg white, and 1.6% based on liquid egg white) fully restored foam expansion and foam liquid stability of 0.4% yolk-contaminated egg white, and it even out-performed the foaming of pure white protein. Therefore, a feasible solution to restore the foaming properties of yolk-contaminated egg white has been identified. It is expected that such modified SPI can be used as an additive or ingredient in foaming formulation, especially when the egg white protein is suspected of lipid contamination.