Physicochemical and bitterness properties of enzymatic pea protein hydrolysates

ABSTRACT:  The effects of different proteolytic treatments on the physiochemical and bitterness properties of pea protein hydrolysates were investigated. A commercial pea protein isolate was digested using each of 5 different proteases to produce protein hydrolysates with varying properties. After 4 h of enzyme digestion, samples were clarified by centrifugation followed by desalting of the supernatant with a 1000 Da membrane; the retentates were then freeze-dried. Alcalase and Flavourzyme™ produced protein hydrolysates with significantly higher ( P < 0.05) degree of hydrolysis when compared to the other proteases. Flavourzyme, papain, and alcalase produced hydrolysates that contained the highest levels of aromatic amino acids, while trypsin hydrolysate had the highest levels of lysine and arginine. Papain hydrolysate contained high molecular weight peptides (10 to 178 kDa) while hydrolysates from the other 4 proteases contained predominantly low molecular weight peptides (≤ 23 kDa). DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging activity of the Flavourzyme hydrolysate was significantly ( P < 0.05) the highest while alcalase and trypsin hydrolysates were the lowest. Inhibition of angiotensin converting enzyme (ACE) activity was significantly higher ( P < 0.05) for papain hydrolysate while Flavourzyme hydrolysate had the least inhibitory activity. Sensory analysis showed that the alcalase hydrolysate was the most bitter while papain and α-chymotrypsin hydrolysates were the least. Among the 5 enzymes used in this study, papain and α-chymotrypsin appear to be the most desirable for producing high quality pea protein hydrolysates because of the low bitterness scores combined with a high level of angiotensin converting enzyme inhibition and moderate free radical scavenging activity.     

Evaluation Of Antioxidant Properties In Vitro of Plastein-Reaction-Stressed Soybean Protein Hydrolysate

Alcalase was used in the present study to carry out an enzymatic hydrolysis of soybean protein isolate and a plastein reaction of the prepared hydrolysate in vitro, aiming to investigate the influence of the plastein reaction on the antioxidant properties of the modified hydrolysate. Soybean protein hydrolysate was prepared in a degree of hydrolysis of 14.0%, exhibited a scavenging activity of 43.6% on ABTS radical in vitro, and thus was used as the substrate of the plastein reaction to prepare the plastein-reaction-stressed hydrolysate. Response surface methodology was applied to select suitable reaction conditions as follows: enzyme addition level 1037 U/g peptides, substrate concentration 29.7% (w/v), reaction temperature 20.3°C. The stressed hydrolysate showed the highest scavenging activity on ABTS radical (about 47.9%) or maximal reaction extent when reaction time was 6 h. Three stressed hydrolysates with different reaction extents were prepared and evaluated for other antioxidant activities. Compared to the original hydrolysate, the stressed hydrolysate with lower reaction extent exhibited a similar (P > 0.05) scavenging activity on DPPH (or superoxide) radical and reducing power, but a significant higher activity (P < 0.05) on hydroxyl radical. The stressed hydrolysate with the highest reaction extent behaved as these investigated antioxidant properties were significantly higher (P < 0.05) than the original hydrolysate except for scavenging activity on DPPH radical. The results of the present study highlight that the alcalase-catalyzed plastein reaction appears to be capable of improving antioxidant properties of soybean protein hydrolysate.     

蛋白含量对牦牛乳清蛋白浓缩物功能特性的影响

通过不同截留分子质量的再生纤维素膜过滤纯化牦牛原乳清液和牦牛甜乳清液,分别制取牦牛原乳清蛋白浓缩物（native whey protein concentrate,NWPC）和牦牛甜乳清蛋白浓缩物（sweet whey protein concentrate,SWPC）,研究蛋白含量不同的乳清蛋白浓缩物（whey protein concentrate,WPC）主要成分（乳糖含量、pH值和总蛋白质含量）和功能特性（溶解性、持水性、持油性、起泡性、乳化性及热稳定性）的特征。结果表明：10 000 Da再生纤维素膜透析得到的牦牛WPC中总蛋白含量达到80%以上,不含乳糖,功能特性（溶解性、持水性、持油性、起泡性、乳化性及热稳定性）均显著高于经3 500 Da卷式膜、5 000 Da再生纤维素膜透析得牦牛WPC,WPC蛋白含量越高,其功能特性越好;不同蛋白含量的牦牛SWPC起泡能力、泡沫稳定性、乳化活性和乳化稳定性均显著（P＜0.05）高于牦牛NWPC。牦牛乳WPC最不稳定温度为85 ℃,高于荷斯坦牛乳WPC的80 ℃,热处理会适当改善牦牛WPC的起泡性能、乳化性能和热稳定性。通过膜牦牛处理获取的高蛋白含量的WPC,功能特性较好,应用广泛,对解决牦牛乳清资源的利用问题、保护环境、提高企业的经济效益起到关键性作用。     

Improving Functional Properties of Soy Protein Hydrolysate by Conjugation with Curdlan

ABSTRACT:  Soy protein hydrolysate was conjugated with curdlan through the naturally occurring Maillard reaction to extend its application in food processing. The gel-forming and emulsifying properties of soy protein hydrolysates were significantly improved ( P < 0.05) by conjugation with curdlan. The soy protein hydrolysate–curdlan conjugate (SPHCC)–soy protein isolate (SPI) mixed gel had a much thicker network than the soy protein hydrolysate and SPI mixed gel judged from scanning electron microscopic images. The improvement of gelling properties of soy protein hydrolysate by curdlan-conjugate was attributed to both the decrease in the repulsive forces among soy protein hydrolysates and the increase in the solubility. The covalent binding of soy protein hydrolysates and curdlan also showed a profound effect on the antioxidative activity of the soy protein hydrolysates. The higher antioxidative activity of SPHCC was related to the peptide reductants produced from the Maillard reaction and the higher emulsifying property of SPHCC. The conjugates of soy protein hydrolysate and curdlan can be used as a functional food additive having excellent gel-forming, emulsifying properties and antioxidative activity.     

响应面试验优化裙带菜蛋白酶解工艺及酶解液抗氧化活性

运用响应面分析方法对裙带菜蛋白酶解工艺条件进行优化。经单酶筛选,在单因素试验基础上,以亚铁离子螯合率和1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率为主要指标,水解度为辅助指标,研究酶解时间、酶解温度、p H值、底物质量浓度、加酶量对裙带菜蛋白酶解产物抗氧化活性和水解度的影响,并比较优化条件下的酶解液与常用天然抗氧化剂抗坏血酸、合成抗氧化剂丁基羟基茴香醚(butyl hydroxyanisole,BHA)的抗氧化活性。结果表明:复合蛋白酶是裙带菜蛋白酶解的最适用酶,酶解液螯合亚铁离子能力和清除DPPH自由基的最优条件为酶解时间8.1 h、酶解温度50℃、p H 7.0、底物质量浓度15 g/L、加酶量0.2%(0.3 AU/g裙带菜粉末)。在此条件下,酶解液的亚铁离子螯合率为88.58%,DPPH自由基清除率为59.22%,水解度为29.72%。对比常用抗氧化剂,在亚铁离子螯合能力方面,酶解液显著高于0.01%抗坏血酸和0.01%BHA(P0.05),而在DPPH自由基清除能力和还原能力方面,酶解液低于0.01%抗坏血酸和0.01%BHA(P0.05)。     

Rheological and Functional Properties of Catfish Skin Protein Hydrolysates

ABSTRACT:  Catfish skin is an abundant and underutilized resource that can be used as a unique protein source to make fish skin protein hydrolysates. The objectives of this study were to isolate soluble and insoluble proteins from hydrolyzed catfish skin, study the rheological and functional properties of the protein hydrolysates, and evaluate the properties of emulsions made from the protein powders. Freeze-dried catfish skin soluble (CSSH) and insoluble hydrolysate (CSISH) powders were analyzed for proximate analysis, emulsion stability, fat absorption, amino acids, color, and rheological properties. CSSH had significantly ( P  < 0.05) higher protein, ash, and moisture content but lower fat content than that of CSISH. The yield of CSSH (21.5%± 2.2%) was higher than that of CSISH (3%± 0.3%). CSISH had higher emulsion stability than CSSH. CSSH was light yellow in color and CSISH was darker. The mean flow index values for emulsion containing CSSH (ECSSH) and CSISH (ECSISH) were both less than 1, indicating that they were both pseudoplastic fluid. The   G '  and   G "  values for the ECSISH were higher than that of ECSSH, indicating that the viscoelastic characteristic of the emulsion containing CSISH was greater than that of the emulsion containing CSSH. The study demonstrated the CSSH and CSISH had good functional and rheological properties. They have potential uses as functional food ingredients.     

Fractionation of Soy Protein Hydrolysates using Ultrafiltration Membranes

The effect of membrane pore size on the molecular weight distribution and selected functional properties of a protein hydrolysate produced from soy isolate and Pronase was examined. Molecular weight distributions were similar for permeates from 5000, 10,000 and 50,000 molecular weight cut-off (MWCO) membranes: two large fractions at 2300 and 1000 daltons. The 100,000 MWCO membrane resulted in three fractions of 25,000, 13,000 and 2300 daltons. Solubility of the hydrolysate increased with decrease in MWCO, while foam stability decreased.     

Enzymatic hydrolysis of blue whiting (Micromesistius poutassou); functional and bioactive properties

Functional and biochemical properties of fish protein hydrolysates (FPH) from blue whiting (BW) were studied. FPH (2.5%, 5%, 10%, and 15% degree of hydrolysis [DH]) were made from isolated proteins from headed and gutted BW with Alcalase 2.4 L. The properties of dried BW mince and protein isolate compared to 4 reference proteins (soy and milk protein) were studied: color, solubility, water-holding capacity (WHC), oil-binding capacity (OBC), emulsion capacity (EC), and emulsion stability (ES). The angiotensin I-converting enzyme (ACE) inhibitory activities of the soluble fraction of BW powders were also investigated. Furthermore, the products were characterized by analyzing their chemical composition. Chemical composition, solubility, OBC, and EC of the BW powders was significantly (P < 0.05) different with different DH, while color, ES, and WHC were not significantly (P > 0.05) different. Salt content of the FPH was high (4% to 19%) and increased with increased DH. Protein solubility varied from 10% to 70% and increased with increased DH. WHC of the FPH was around 97% and was higher than that of all the reference proteins tested. OBC decreased with increased DH (from 3.5 to 2.1 g oil/g protein) and was higher than OBC of the soy and milk proteins (1.6 to 1.9 g oil/g protein). EC of FPH was similar or lower than the reference proteins. ES of FPH (60% to 90%) was similar to or lower than soy and whey proteins (60% to 98%) but higher than casein (20%). ACE inhibition activity increased as DH was increased. Practical Application: The results from this study demonstrate that a functional bioactive hydrolysate can be produced from BW, which is an underutilized fish species, and may aid the industry in better utilizing this raw material. The novelty of this research was the use of BW as a raw material where the protein has been isolated with the pH shift method. Furthermore, it was novel that bioactivity and functionality was measured in the same samples.     

Emulsifying properties of soy protein isolates obtained by microfiltration

Soy protein isolate (SPI) fractions were produced using two different pore size microfiltration membranes. Microfiltration was carried out on SPI produced by isoelectric precipitation of a crude protein extract. Five fractions were obtained: two retentates and two permeates from the two membranes plus an intermediate fraction obtained as the retentate on the small‐pore‐size membrane using the permeate from the larger‐pore‐size membrane. Emulsions stabilised by the retentate fractions exhibited higher values (P < 0.01) of emulsion stability index (ESI) and emulsifying activity index (EAI) than those stabilised with fractions made from the permeates. The intermediate fraction gave intermediate ESI values, while the EAI values were not significantly different from those for SPI and one of the retentates. SDS‐PAGE profiles indicated that the fractions exhibiting high functionality in terms of ESI and EAI were also richer in 7S globulin soy protein subunits. © 2002 Society of Chemical Industry     

Functional properties of soy protein hydrolysate produced from a continuous membrane reactor system

Two types of soy protein hydrolysates were produced by hydrolysis of isolated soy protein (ISP) with a combination of Alcalase and Flavourzyme in a continuous membrane system with 3000 and 30,000 daltons molecular-weight-cut-off (MWCO) membranes, respectively. Both hydrolysates were completely soluble over a pH range of 2–9. Their water-binding capacity increased 1.8–3.4 times at a water activity of 0.6–0.95 as compared to intact ISP. The antioxidant activity of ISP was remarkably enhanced by enzyme hydrolysis. The hydrolysate from the 30,000 daltons MWCO membrane had a much higher antioxidant activity than that from the 3000 daltons MWCO membrane. The results suggested that both selected enzymes and MWCO membrane governed the functional properties of protein hydrolysates. Long-term operation study showed that the membrane reactor system could maintain a steady production of ISP hydrolysate over 16 h.     

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。     

Ultrasonic assisted alkali extraction of protein from defatted rice bran and properties of the protein concentrates

Alkaline extraction for the preparation of protein concentrate from rice bran was compared with a range of ultrasonic treatments. Results revealed that the extraction time decreased, and the reaction rate constant increased, with increasing ultrasonic power. The reaction rate constants were 0.0065, 0.0130, 0.0237 and 0.0924 at 40, 60, 80 and 100 W respectively. The defatted rice bran protein concentrate (DRBPC) using ultrasonication (100 W for 5 min) and conventional methods showed no significant difference in bulk densities ( P  > 0.05) but it had higher yield (%) and was lighter brown using ultrasonication ( P  ≤ 0.05). The SEM showed that the residual rice bran after extracting protein using ultrasonication exhibited more damage than the conventional method. The functional properties of both samples were not significantly different ( P  > 0.05) in terms of foam and emulsifying stability. However, the water and oil absorption, foam capacity and emulsion activity were significantly different ( P  ≤ 0.05). The nitrogen solubility index of both DRBPC samples gave similar profiles with the lowest solubility at pH 4–6.     

食盐对猪肉脱水过程中物理特性及微观结构的影响

探究食盐用量对脱水腌制肉的色泽、水分活度、体积变化率、横向收缩比、纵向收缩比等物理特性、微观结构变化的影响,分析各指标间的相关性。结果表明：食盐用量主要影响脱水肉样的a*、L*值,可减少肉色变化,但效果有限;食盐用量在1%～8%之间时,用量越高干燥过程中肉样水分活度随水分减少下降越快,超过8%后影响减弱;1%食盐用量腌制肉样体积变化率明显高于其他各样（P＜0.05）,4%食盐用量腌制肉样的纵向收缩比显著小于其他腌制样（P＜0.05）,而食盐用量对腌制样横向收缩比无显著影响（P＞0.05）;脱水腌制肉样水分活度、体积变化率、横向收缩比及纵向收缩比、含水率之间的相关关系不受食盐用量的影响;腌制液食盐用量越高,脱水肉样肌纤维及其膜的变化越明显。     

酶解大豆分离蛋白乳化特性的研究

利用枯草芽孢杆菌AS1．398中性蛋白酶对大豆分离蛋白进行水解，并利用浊度法测定了不同水解度、不同pH条件下酶解大豆分离蛋白的乳化特性，结果表明：AS1．398蛋白酶对大豆分离蛋白的最大水解度为36％，水解度为9％时乳化活性最大，水解度为3％是乳化稳定性最好。同一水解度时，pH越高，蛋白质的乳化特性越好。水解度为3％、9％、15％的大豆分离蛋白在pH等于或高于5．0时的乳化活性明显地高于原蛋白质，且水解度为3％时乳化稳定性也明显地高于原蛋白质。     

Functional Properties of Hydrolysates from Proteolysis of Heat-denatured Whey Protein Isolate

Heat-denatured whey protein isolate was hydrolyzed with trypsin, α-chymotrypsin, Alcalase or Neutrase to 2.8, 4.3, 6.0 or 8.0% degree of hydrolysis. Hydrolysates were fractionated by ultrafiltration and freeze-dried. Protein content of retentates showed little variation but permeates differed with enzyme. Surface hydrophobicity increased with hydrolysis but was not linear except for α-chymotrypsin. Ultrafiltration increased solubility and the permeates and retentates had better solubility than hydrolysates. Retentates had higher emulsifying activity index than hydrolysates while permeates did not form stable emulsions. Permeates formed stable foams but hydrolysates and retentates showed poor foaming characteristics. Specificity of the enzyme, and degree of hydrolysis influenced the functional properties of the peptides. Fractions generated by trypsin, at all levels of hydrolysis generally had higher solubility, emulsifying properties and foaming properties. Permeates from Alcalase hydrolysis had the best foam capacity but low foam stability.     

Influence of degree of hydrolysis on functional properties,antioxidant activity and ACE inhibitory activity of peanut protein hydrolysate

Functional properties, antioxidant and Angiotensin-converting enzyme (ACE) inhibitory activities of peanut protein isolate (PPI) and peanut protein hydrolysate (PPH) prepared using Alcalase, at different (10%, 20%, 30% and 40%) degrees of hydrolysis, (DH) were investigated. Hydrolysis (at DH > 10%) significantly (p < 0.05) improved the solubility (>80%) of PPI, especially in the pH range of 4–6. However, PPI showed better emulsifying and foaming properties than PPH (p < 0.05). As DH increased, ferrous ion chelating activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity and ACE inhibitory activity of PPH increased, while reducing power decreased (p < 0.05). Bleaching of beta-carotene by linoleic acid was suppressed better by PPI and PPH at 10% DH than of PPH at higher DH. Thus, the results reveal that DH affects functional properties, antioxidant and ACE inhibitory activities of peanut protein.     

油菜蜂花粉ACE抑制活性酶解物的制备及分离

采用4 种蛋白酶水解油菜蜂花粉蛋白制备ACE 抑制活性物质,高效液相色谱法测定油菜蜂花粉蛋白水解物对ACE 的抑制率。结果表明：油菜蜂花粉蛋白酶水解物具有ACE 抑制活性,水解物对ACE 的抑制活性差异显著(P ＜ 0.05),其中碱性蛋白酶＞中性蛋白酶＞木瓜蛋白酶＞酸性蛋白酶,碱性蛋白酶水解物的IC50 为0.35mg/mL。4 种蛋白酶水解物经Bio P-2 凝胶分离后,ACE 抑制活性较强的组分主要集中在保留时间70～120min,在此区间碱性蛋白酶水解物分离组分对ACE 的抑制率达到90% 以上,分子质量在376.4～1355D 之间。     

乳清多肽对猪肉糜氧化和凝胶特性的影响

目的：研究乳清多肽对猪肉糜氧化和凝胶特性的影响作用。方法：实验分为6 组,第1组为空白对照组,第2组加入20%（质量分数,下同）的乳清分离蛋白未水解物,第3～5组中分别加入10%、15%、20%的乳清分离蛋白水解物冻干粉,第6组中加入0.02%的丁基羟基茴香醚（butylated hydroxyanisole,BHA）。在肉糜冷藏7 d过程中测定过氧化值、羰基含量、巯基含量及凝胶的质构、白度、保水性、流变学特性的变化。结果：在贮藏初期,处理组和对照组之间无显著差异（P＞0.05）。贮藏7 d后,添加乳清分离蛋白水解物组在抑制氧化和保护凝胶品质方面都要比对照组和未水解组好（P＜0.05）。其中添加20%乳清分离蛋白水解物冻干粉的储能模量的极大值最高,而在抑制脂肪、蛋白氧化及保护凝胶质构、保水性方面,添加15%乳清分离蛋白水解物冻干粉效果更好（P＜0.05）,接近甚至达到了BHA组水平。结论：乳清多肽具有抑制猪肉糜氧化和保护肉糜凝胶品质的作用。     

对羟基苯甲醛和阿魏酸对酿酒酵母理化特性的影响

研究木质纤维素水解液中的酚类物质对羟基苯甲醛和阿魏酸对酿酒酵母（Saccharomyces cerevisiae）GGSF16理化特性的影响。结果表明,经药物处理后酿酒酵母的糖代谢时间和乙醇发酵时间延长,胞外核酸、蛋白质和胞内海藻糖含量均显著增加（P＜0.05）,扫描电子显微镜图可以看出,原本完整光滑的细胞经对羟基苯甲醛和阿魏酸处理后细胞表面出现褶皱、粘黏、裂解现象,结合傅里叶变换红外光谱呈现的代表脂质、蛋白质、核酸等分子成分官能团的改变可以得出,对羟基苯甲醛和阿魏酸能够破坏酿酒酵母的细胞壁,增大细胞膜的通透性,从而使细胞内容物流出,导致细胞损伤。