Rheological properties of soy protein hydrolysates obtained from limited enzymatic hydrolysis

Soy protein products hexane-defatted soy flour, extruded-expelled soy flour, soy protein concentrate and soy protein isolate, were modified by using the enzyme bromelain to 2% and 4% degrees of hydrolysis (DH). Peptide profiles, water solubility, and rheological properties including dynamic shear, large deformation, and apparent viscosities of resulting hydrolysates were determined. Protein subunits profiles for the hydrolysed isolates and concentrates were extensively altered by the treatment while only minor changes were observed for the hydrolysed flours. Water solubility profiles of all hydrolysates in the pH range of 3.0-7.0 were enhanced by hydrolysis. For the unhydrolysed controls, the isolate had the highest storage modulus (G′), followed by the concentrate, the extruded-expelled flour and the hexane-defatted flour. The hydrolysates retained some of their gelling ability even though the losses in storage modulus (G′) were substantial. After heating step to 95 °C, the G′ values of all substrates at 25 °C decreased with increase in DH. Texture profile analyses of the soy protein gels were also lower in hardness after hydrolysis. The Power Law model provided excellent fit to hydrolysate dispersions flow (R²>0.99). Hydrolysis decreased the consistency coefficients of dispersion and increased flow behavior index resulting in thinner dispersions. These results suggest that limited protease hydrolysis of various soy protein meals with bromelain produce soy protein ingredients with modified rheological properties.     

Egg yolk protein modification by controlled enzymatic hydrolysis for improved functionalities

Delipidated egg yolk protein (EYP) is produced as a co-product of egg yolk lecithin extraction. This EYP is expected to have poor functionality because of protein denaturation caused by ethanol treatment during lecithin extraction. Two food grade endo-proteases were used to produce EYP hydrolysates (EYPh) with two degrees of hydrolysis (DH), 3% and 6%. Protein solubility was improved as DH increased. Solubility profiles for both EYP and EYPh were relatively less pH-dependent compared with soy protein. Except for foaming capacity, EYPh showed improvement in foam stability, foaming speed, and foam density. Emulsion stability was improved for all EYPhs. Treatments at DH of 6% showed significant increase in emulsification capacity. We have shown quantitatively how controlled enzymatic hydrolysis can be applied to ethanol-treated lipid-free EYP to increase protein solubility, and thus to improve foaming and emulsification properties.     

Some functional properties of fractionated soy protein isolates obtained by microfiltration

Five soy proteins isolate (SPI) fractions were produced using two microfiltration membranes with different pore sizes. Fractionation was carried out on SPI produced by isoelectric precipitation of a crude protein extract. The five fractions were two retentates and two permeates from the two membranes, the fifth fraction was obtained as the retentate on the smaller-pore-sized membrane fed with the permeate from the larger-pore-sized membrane. Solubility, foaming and emulsifying properties of the collected fractionates were investigated. It was observed that in the pH range 3–8 the retentates featured superior solubility compared with permeates. There was no significant difference (p>0.01) in solubility between the retentates and SPI at pH6. Foaming characteristics of the fractions followed the same trend as solubility with regard to foam expansion. There was, however, no particular trend observed with regards to foam stability. Emulsions stabilised by the retentates exhibited higher values (p<0.01) of emulsion stability index (ESI) and emulsifying activity index (EAI) than those stabilised with permeates. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) profiles indicated that the fractions exhibiting high functionality in terms of solubility, foaming and emulsifying properties were also richer in 7S globulin soy protein subunits.Isoelectric focussing (IEF) profiles showed that retentates were richer in species with isoelectric points (pI) between 5.2 and 5.6 while permeates featured more prominently at pIs between 4.5 and 4.8.     

不同酶切方式对乳清蛋白疏水性和乳化性的影响

采用不同的蛋白酶对乳清蛋白进行水解,考察了肽键断裂方式对乳清蛋白肽疏水性和乳化性的影响,以及乳清蛋白不同酶解产物的疏水性和乳化性的关系。结果表明:不同蛋白酶作用于乳清蛋白得到的水解产物疏水性不同,6种蛋白酶解液的疏水性均随水解度的增大而降低,其中以胰凝乳蛋白酶酶解液的疏水性下降的最慢。研究还发现,乳化性随着水解度增加而先升高后下降,以双酶复合酶解液最差。不同蛋白酶水解液的乳化性指数随疏水性指数的降低而升高,乳化性指数与疏水性氨基酸质量分数成正相关。     

酶水解法提高大豆蛋白水解度的研究

用蛋白酶水解方法提高大豆分离蛋白水解度.通过对多种蛋白酶的对比分析可知,风味蛋白酶的水解效果好于其它蛋白酶,但正交试验结果表明,即使在优化条件下水解,单一的风味蛋白酶水解所得大豆分离蛋白水解度最高只达到43.12%.若先用胃蛋白酶水解再用风味蛋白酶水解则水解度最高可达68.81%,而风味蛋白酶与其它酶的联合应用效果略差;若先使用风味蛋白酶后使用胃酶则水解度只有47.89%.表明不同酶对大豆蛋白分子具有不同的水解特点.     

酶解对蛋清蛋白液起泡性能和乳化性能的影响

利用碱性蛋白酶水解蛋清蛋白,主要研究了酶用量、底物浓度和酶解时间对蛋清蛋白液起泡性能和乳化性能的影响。实验结果表明,酶解条件对蛋清蛋白液的起泡性能和乳化性能有一定程度的影响,当碱性蛋白酶酶解温度为45~50℃,pH6~8.5时,其最佳酶解条件为酶用量4%,底物浓度3%,酶解时间6h。在此最佳酶解条件下获得的蛋清蛋白酶解液起泡性为27.65%,发泡能力178.26,泡沫稳定性60.33%,乳化容量37.86%,乳化稳定性127.23,乳化活性指数134.53,乳状液稳定指数1.83。      

Preparation and characterization of bio-nanocomposite films based on soy protein isolate and montmorillonite using melt extrusion

The non-biodegradable and non-renewable nature of plastic packaging has led to a renewed interest in packaging materials based on bio-nanocomposites (biopolymer matrix reinforced with nanoparticles such as layered silicates). Bio-nanocomposite films based on soy protein isolate (SPI) and montmorillonite (MMT) were prepared using melt extrusion. Effects of the pH of film forming solution, MMT content, and extrusion processing parameters (screw speed and barrel temperature distribution) on the structure and properties of SPI–MMT bio-nanocomposite films were investigated. X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) were used for structural characterization of the films. Properties of the films were determined by tensile testing, dynamic mechanical analysis (DMA), thermogravimetric analysis (TGA), and water vapor barrier measurement. The arrangement of MMT in the soy protein matrix ranged from exfoliated at lower MMT content (5%) to intercalated at higher MMT content (15%). There was a significant improvement in mechanical (tensile strength and percent elongation at break) and dynamic mechanical properties (glass transition temperature and storage modulus), thermal stability, and water vapor permeability of the films with the addition of MMT. The results presented in this study show the feasibility of using bio-nanocomposite technology to improve the properties of biopolymer films based on SPI.     

Improved emulsifying properties of soy proteins by acylation with saturated fatty acids

Effects of acylation on emulsifying properties of soy proteins were investigated using a variety of saturated fatty acids. Beta conglycinin (7S), glycinin (11S), and acid-precipitated protein (APP) were acylated with activated fatty acid esters (6C–18C) to form covalent linkage between the carboxyl group of the fatty acid and the free amino groups of the protein. Reduction in the free amino groups of acylated 7S, 11S and APP resulted into the dissociation of the protein, indicating a structural change, as evidenced by the fluorescence spectra and the degree of modification. It was shown that the emulsifying activity (EAI) and emulsion stability (ES) of 7S and 11S were significantly improved (p < 0.05) upon acylation with all saturated fatty acids, whereas no change in EAI and ES for the acylated APP was observed upon attachment of short and long chain fatty acids. The fluorescence intensity was also remarkably affected by acylation showing significant changes in protein structure. Covalent attachment of fatty acids resulted into 1.4–2.2 and 1.1–1.8-fold increase in the oil binding capacity (OBC) of 7S and 11S respectively, however no changes in acylated APP. Acylated 7S showed 3.0–9.4-fold increase in the water binding capacity (WBC), with no change in acylated 11S, while acylated APP with longest chains showed low WBC. The surface hydrophobicity of 7S was significantly improved (p < 0.05) by acylation; no changes were observed in the acylated 11S. Furthermore, acylation decreased the surface hydrophobicity of APP. Thus, it was demonstrated that saturated fatty acids with adequate chain length are suitable candidates for the preparation of functional lipoproteins from soy proteins.     

低限度酶水解对醇法大豆浓缩蛋白分散性影响

以大豆浓缩蛋白分散时间和分散稳定时间为主要指标,研究大豆浓缩蛋白经过Alcalase酶低限度酶水解后,分散时间和分散稳定时间变化情况,并考察低限度酶水解对分子量分布影响。     

Influence of the extent of enzymatic hydrolysis on the functional properties of protein hydrolysate from grass carp (Ctenopharyngodon idella) skin

Protein hydrolysates from grass carp skin were obtained by enzymatic hydrolysis using Alcalase^®. Hydrolysis was performed using the pH-stat method. The hydrolysis reaction was terminated by heating the mixture to 95 °C for 15 min. At 5.02%, 10.4%, and 14.9% degree of hydrolysis (DH), the hydrolysates were analyzed for functional properties. The protein hydrolysates had desirable essential amino acid profiles. Results demonstrated that the hydrolysates had better oil holding and emulsifying capacity at low DH. The water holding capacity increased with increased levels of hydrolysis. Enzymatic modification was responsible for the changes in protein functionality. These results suggest that grass carp fish skin hydrolysates could find potential use as functional food ingredients as emulsifiers and binder agents.     

限制性酶解乳清蛋白功能性质研究

探究乳清蛋白在碱性蛋白酶限制性水解下功能性质变化。以乳清蛋白的溶解性,乳化性、乳化稳定性,起泡性、起泡稳定性为考察指标,确定乳清蛋白的等电点及分析不同水解度下乳清蛋白功能性质在p H调控下的变化。结果表明:乳清蛋白的等电点为4.8。乳清蛋白进行限制性酶解后功能性质有了很大提高,其中溶解性在DH14、p H10下达到最大值,较原蛋白提高了14.55%;起泡性在DH14、p H4下达到最大值,较原蛋白提高了107.5%;起泡稳定性在DH4、p H4下达到最大值,比原蛋白提高了8.66%;乳化性在DH14、p H12下达到最大值,比原蛋白提高了56.1%;乳化稳定性DH4、p H12下达到最大值,比原蛋白提高了50.42%。      

Functional properties of Maillard reaction products of rice protein hydrolysates with mono-, oligo- and polysaccharides

The Maillard reaction was carried out under wet reaction conditions using rice protein product from limited hydrolysis to conjugate with various mono-, oligo- and polysaccharides. The Maillard reaction products (MRPs) prepared with rice protein hydrolysates at 5% degree (hydrolysed by Protease N) and dextran T20 (20 min at 100 °C) showed the greatest improvement in functionality. The solubility, emulsification activity (EA), and emulsification stability (ES) of the MRPs increased by factors of 3.5, 5.3 and 7.3 times, respectively as compared to MRPs formed with native rice proteins and dextran T20. Amino acid analysis indicated that lysine and arginine decreased significantly in the MRPs. Formation of MRPs reduced the surface hydrophobicity, which was in agreement with the change of solubility. However, further decreasing of the surface hydrophobicity resulted in lower EA and ES during the late stage of the reaction. Fluorescence analysis suggested that formation of late stage MRPs occurred after 20 min of the Maillard reaction. The molecular weight distributions showed that the functional properties of MRPs and the mechanisms of formation were affected by the peptide chain length.     

The effect of enzymatic hydrolysis time and temperature on the properties of protein hydrolysates from Persian sturgeon (Acipenser persicus) viscera

Protein hydrolysate was prepared from the viscera of Persian sturgeon (Acipenser persicus), a major sturgeon species in the Caspian Sea. Hydrolysis was performed at three different temperatures (35, 45 and 55 °C), pH 8.5, using commercially available Alcalase^® and an enzyme to substrate ratio of 0.1 AU/g viscera protein over a 205 min incubation period. Protein and lipid content of the hydrolysate were 65.82%, and 0.18%, respectively. Protein recovery and degree of hydrolysis ranged from 34.97% to 61.96% and 13.32% to 46.13%, respectively. The highest degree of hydrolysis was observed at 55 °C after 205 min (p < 0.05). The amino acid score of the hydrolysates was similar to that of the FAO/WHO reference protein. It is revealed that Persian sturgeon visceral protein hydrolysate amino acid fulfils adult human requirements. Based on National Research Council guidelines, phenylalanine is the first limiting amino acid in the hydrolysate. However, the hydrolysate has the potential for application as an ingredient in formulated diets.     

不同预处理方法对制备水解植物蛋白的影响

为在酶解制备水解植物蛋白时降低成本,同时保持较高的水解度及鲜味氨基酸含量,采用不同方法对原料进行前处理,使蛋白质变性,暴露酶的作用位点.加速酶解的进行.研究表明:热变性处理中,沸水浴处理 20 min 及微波处理 40 s,水解度分别提高 7%、5%.116℃下的高温蒸气作用 45 min,水解度可以提高 11%,而且生成谷氨酸的量也最高.超声前处理对蛋白质酶解的影响不是很明显,仅能提高2%.原料水分含量对蛋白质变性有较大的影响.热前处理时,水分含量80%可以较有效地提高蛋白酶解的水解度.     

转谷氨酰胺酶交联木瓜蛋白酶水解产物改善大豆分离蛋白乳化特性的研究

为获得适宜添加到冰淇淋等冷冻食品中的大豆分离蛋白（SPI）,利用转谷氨酰胺酶交联木瓜蛋白酶酶解大豆分离蛋白产物来改善大豆蛋白的乳化特性。将样品经高压均质制成O/W型乳状液,研究不同水解程度的水解产物交联后对样品乳状液的粒度分布、表面积平均粒径、离心乳析率等的影响,同时研究了常温放置、低温放置（4℃）以及冷冻-解冻处理对酶改性蛋白乳状液聚集、聚结稳定性的影响。结果表明,SPI和中低度水解（DH2%¹0%）样品经转谷氨酰胺酶交联后乳化活性和乳化稳定性均显著提高,水解度10%样品交联后的乳化活性和乳化稳定性改善最显著。酶改性可以显著改善SPI新鲜乳状液的聚集稳定性,中度水解度（DH6%¹0%）SPI样品交联后聚集程度最低。冷藏不会明显损坏样品的聚集和聚结稳定性。冷冻-解冻处理使乳状液发生聚结现象,SPI乳状液的聚结程度最大,中度水解度（DH6%¹0%）样品交联后的聚结程度最小。因此,可将中度水解SPI交联样品用于对乳化特性要求较高的冰淇淋等冷冻食品中。      

Transglutaminase cross-linking to enhance elastic properties of soy protein hydrogels with intercalated montmorillonite nanoclay

To strengthen the network of bio-nanocomposite hydrogels, layered montmorillonite (MMT) nanoclay was intercalated by surface-coating with soy protein (SP) before mixing with 6% w/v SP for cross-linking by microbial transglutaminase (mTGase). Dynamic rheology was performed to study variables of NaCl and mTGase concentrations, with and without 1% w/v MMT. Without mTGase, the highest storage modulus (G′) was observed at 100 mM for samples without MMT, which was twice of the highest G′ for samples with MMT, at 200 mM NaCl. With mTGase, a shorter gelation time and a stronger hydrogel were observed at a higher enzyme level. Overall, the non-gelling 6% w/v SP dispersion was transformed to a hydrogel with G′ of 1099 Pa after addition of 100 mM NaCl and 1% SP-coated MMT and treatments by 6.25 U/g-protein mTGase for 2 h and heating/cooling steps. The integration of surface-coating and mTGase cross-linking is promising to improve properties of the nanocomposite system.     

超声处理对绿豆分离蛋白乳化性的影响

研究超声处理对绿豆分离蛋白乳化性的影响,对不同超声功率、超声时间、底物蛋白浓度对绿豆分离蛋白乳化性的影响进行分析,并通过响应面实验,最终得到超声处理提高绿豆分离蛋白乳化性的最佳条件。结果表明,超声处理提高绿豆分离蛋白乳化性的最佳条件为:超声功率410 W、超声时间10.5 min、底物蛋白浓度11.5%。该处理条件下得到改性后的绿豆分离蛋白乳化性为35.01 m2/g。      

Effects of the proteinaceous moiety on the emulsifying properties of sugar beet pectin

The role of the proteinaceous moiety in emulsifying was investigated using pectin from sugar beet as a model polysaccharide. Physicochemical and macromolecular characteristics of sugar beet pectin were examined with or without an enzymatic modification using multiple acid-proteinases. The enzymatic modification decreased the total protein content from 1.56±0.15% to 0.13±0.02% by the Bradford method without significant change in ferulic acid or most constitutional sugars. It also decreased the weight-average molecular weight (M_w) from 517±28 to 254±20 kg/mol and the z-average root-mean-square radius of gyration from 43.6±0.8 to 35.0±0.6 nm. Emulsifying properties of the polysaccharide with or without the enzymatic modification were evaluated by emulsion droplet size and creaming stability of O/W emulsions (pH 3.0) containing 15 w/w% middle-chain triglyceride and 1.5 w/w% sugar beet pectin as main constituents. The modification increased the average diameter (d_3,2) of emulsion droplets from 0.56±0.04 to 3.00±0.25 μm immediately after the preparation, suggesting a decrease in the emulsifying activity. It caused the creaming of the emulsions during incubation at 60 °C, which was in line with the finding that macroscopic phase separation occurred only in the presence of the modified pectin after storage at 20 °C for a day, suggesting a decrease in the emulsion stabilizing ability. The modification also decreased significantly the amount of the pectin fraction that adsorbed onto the surface of emulsion droplets from 14.58±2.21% to 1.22±0.03% and the interfacial concentration of the polysaccharide from 1.42±0.23 to 0.45±0.05 mg/m², where the proteinaceous materials in the pectin molecules activated the oil-water interface. Results from the present study suggest an important role of the proteinaceous moiety to explain the emulsifying properties of sugar beet pectin as in the case of gum arabic and soy soluble polysaccharide.     

Functional properties of safflower protein isolates: Water absorption,whipping and emulsifying characteristics

Safflower meal defatted at the laboratory (SML) and safflower cake provided by an industrial plant (SMI) were used for protein isolation. Isolates were obtained by micellisation (MP) and isoelectric precipitation (IP) techniques. The functional properties studied with these isolates were water and fat absorption, foam expansion and stability, and emulsion activity and stability. Within the experimental conditions of this research, it was observed that the various treatments received previously by the safflower meal, for oil extraction, seemed to have comparatively minor effects on the functional performance of the protein isolates. Only in the case of SML, did the MP isolates have a higher water and fat absorption, and foam expansion than IP samples. For both SML and SMI, the emulsion stability of MP was higher than that of IP.     

Effect of whey protein enzymatic hydrolysis on the rheological properties of acid-induced gels

A protein dispersion blend of β-lactoglobulin and α-lactalbumin was heat-denatured at pH 7.5, hydrolyzed by α-chymotrypsin and then acidified with glucono-δ-lactone to form gels at room temperature. Heat treatment induced the formation of whey protein polymers with high concentration of reactive thiol groups (37 μmol/g). The reactive thiol group concentration was reduced by half after 40 min enzymatic hydrolysis. It was further reduced after enzyme thermal deactivation. During acidification, the first sign of aggregation for hydrolyzed polymers occurred earlier than for non hydrolyzed polymers. Increasing the hydrolysis duration up to 30 min resulted in more turbid gels characterized by an open microstructure. Elastic and viscous moduli were both reduced, while the relaxation coefficient and the stress decay rate constants were increased by increasing the hydrolysis duration. After one week storage at 5 °C, the hardness of gels made from hydrolyzed polymers increased by more than 50%. The effect of polymer hydrolysis on acid-induced gelation is discussed in relation to the availability and reactivity of thiol groups during gel formation and storage.