优选南极磷虾蛋白肽抗氧化活性组分

目的制备南极磷虾抗氧化肽,优选出抗氧化活性最好的南极磷虾肽成分。方法采用脱脂、酶解、超滤等手段制备南极磷虾抗氧化肽;以DPPH自由基清除率、ABTS自由基清除率、抗氧化能力指数3个抗氧化指标和超氧化物歧化酶(superoxide dismutase, SOD)、过氧化氢酶(catalase, CAT)活力2个酶活力指标为评价指标,优选出抗氧化活性最好的南极磷虾肽组分;基于G-25凝胶层析技术对抗氧化活性最好的南极磷虾肽组分进行分离纯化,进一步基于电子顺磁共振(electron paramagnetic resonance,EPR)方法测定洗脱后各峰的DPPH自由基清除率,得到抗氧化活性最好的南极磷虾抗氧化肽组分。结果通过抗氧化指标测定,截留分子量3～10 KDa的肽的DPPH自由基清除率最高,为(30.10±1.10)%,截留分子量3 KDa的南极磷虾肽的ABTS清除能力和抗氧化指数较好,则IC50值为(0.74±0.08) mg/mL、氧化自由基吸收能力(oxygen radical absorbance capacity, ORAC)值为6.39±0.21;通过酶活力指标测定,截留分子量3 KDa的肽的SOD活力和CAT活力最好,分别为(45.7±0.13)U/mg和(17.1±0.19)U/mg蛋白质。对截留分子量3KDa和3～10KDa的南极磷虾肽进行G-25分离纯化后,测定各组分的DPPH自由基清除率,可知截留分子量3～10KDa的F2-2峰清除率最好,为(51.55±1.54)%。结论基于EPR方法优选出分子量为3～10 KDa的南极磷虾肽的F2-2组分的DPPH自由基清除率最高。     

菌酶联合制备猪血抗氧化低聚肽

以猪血为原料,依次采用枯草芽孢杆菌发酵和碱性蛋白酶水解制备猪血抗氧化低聚肽,通过响应面试验优化发酵工艺和酶解工艺,最佳发酵参数为:底物浓度59.0 g/L、发酵时间56.5 h、接种量3.22∶100(体积比),此时多肽含量为2.31 mg/mL,·OH清除率为78.94%;最佳酶解参数为:酶解时间3.0 h、pH 9.5、酶解温度60℃、酶底比390 U/g,此条件下制备得到酶解液的多肽含量5.48 mg/mL,多肽含量较单一发酵提高2.39倍,·OH清除率为93.62%。采用超滤分离法,获得分子量分别为0~1 kDa、1 k Da~5 kDa和0~5 kDa的猪血抗氧化低聚肽,采用7种体外抗氧化指标(·OH清除率、·O~(2-)清除率,抑制脂质过氧化能力、还原力、ABTS~+·清除率、DPPH·清除率、总抗氧化力)评价3种不同分子量段的猪血低聚肽的抗氧化活性,结果表明:3种不同分子量段抗氧化肽活性大小为0~1 kDa0~5 kDa1 kDa~5 kDa,提示高抗氧化活性的猪血低聚肽的分子量集中在1 kDa以下。     

鲜马奶乳清蛋白抗氧化肽分离纯化及其活性研究

分离纯化鲜马奶抗氧化活性肽并检测其抗氧化活性。以胰蛋白酶为酶源酶解鲜马奶乳清蛋白,采用3KDa、10KDa、30KDa不同分子量范围超滤膜对水解液进行分离,分子量在3~10KDa段的酶解液还原能力、DPPH清除能力、羟自由基清除能力,超氧阴离子清除能力均较高,分别为0.8949、63.19%、77.13%、70.08%。用SephadexG-50葡聚糖凝胶色谱纯化鲜马奶抗氧化活性肽并检测其抗氧化活性,得到A、B两个组分,B组分的还原能力、DPPH清除能力、羟自由基清除能力、超氧阴离子清除能力均高于A,分别为0.9814、55.28%、74.25%、56.67%。说明鲜马奶乳清蛋白酶解液具有一定的抗氧化活性,为鲜马奶抗氧化肽进一步纯化、结构鉴定及其生物活性研究提供基础。     

银耳多糖湿法打浆快速提取工艺的研究

以银耳子实体为原料,研究湿法打浆快速提取银耳多糖的工艺条件。以银耳多糖提取率为评价指标,考察料液比、打浆时间、加水温度3个影响因素对银耳多糖提取率的影响,并通过正交试验,确定出最佳提取条件。使用凝胶渗透色谱法(GPC)测定银耳多糖的分子量。结果表明:最佳优化条件为料液比1∶50(g∶m L),打浆时间7min,加水温度为80℃,在此最佳条件下银耳粗多糖的提取率可达40.57%,是传统热水浸提法多糖提取率的2.2倍,提取时间仅为传统热水浸提的1/60。湿法打浆快速提取法和热水浸提法所制备的银耳多糖的平均分子量分别为5.89×10~4KDa和5.86×10~4KDa。     

不同分子量真鲷鱼骨多肽抗氧化活性的研究

以真鲷鱼骨为原料,经胰蛋白酶、风味蛋白酶酶解后,用截留分子量分别为10、5、3ku的超滤膜将其分离成4个分子量段,研究了不同分子量段对·OH、DPPH·和O2-·的清除能力,对Fe3+还原能力以及对油脂的抗氧化性。结果表明:不同分子量多肽组分均具有抗氧化活性,其抗氧化活性强弱为:分子量小于3ku多肽组分>分子量在3~5ku多肽>分子量在5~10ku多肽>分子量大于10ku多肽。     

斑鰶鱼蛋白控制酶解及其酶解物抗氧化活性研究

以斑鰶鱼(Clupanodom punchtatus)为原料,采用食品级的风味蛋白酶、复合蛋白酶、碱性蛋白酶、胰酶和木瓜蛋白酶酶解斑鰶鱼蛋白,制备具有DPPH自由基清除活性的富肽酶解物;对酶解物在化学反应体系和亚油酸氧化体系中的抗氧化活性进行综合评价。研究结果表明:碱性蛋白酶为斑鰶鱼蛋白制备抗氧化肽的最适酶,最优制备条件是:温度55℃,酶解时间5h,料水比1∶3,加酶量0.2%,pH 7.5,此时酶解物(蛋白质量浓度10 mg/mL)DPPH自由基清除率为89.54%;在化学反应体系中,酶解物的DPPH自由基清除能力、还原力和亚铁离子螯合力都随着蛋白质量浓度的增加而增强。在蛋白质量浓度为5 mg/mL的条件下,DPPH自由基清除率为78.26%;酶解物的还原力为0.372(低于抗坏血酸);亚铁离子螯合率为46.81%。在亚油酸氧化体系中,酶解物(蛋白质量浓度10 mg/mL)对亚油酸过氧化物的抑制率为59.61%。     

酶水解猪血红蛋白过程蛋白质降解和氨基酸释放

采用商业酶(Pancreatin,Protamex,Pancreatin/Protamex)水解肉品加工业的副产品猪血血红蛋白.水解后的血红蛋白产物采用凝胶过滤色谱(GPC)及HPLC分析水解过程蛋白质降解及氨基酸释放规律.Protamex水解产物具有宽泛的分子量分布且含有大量的大分子肽(＞15kDa);Pancreatin水解产物同样具有宽泛的分子量分布,但含有大量的小分子肽(＜5kDa).氨基酸组成分析结果表明,Pancreatin酶和Protamex酶均对疏水性氨基酸有较强的作用能力,而且这两种酶的作用住点有互补效应.Pancreatin/Protamex水解产物具有最为复杂的分子量分布图谱,在酶解24h后.水解得到最高的蛋白质回收率为92.34%.在Pancreatin/Protamex水解后期,产物中的肽尤其是大分子肽(＞15kDa)和含有较多疏水性氨基酸的肽容易聚集沉降,导致蛋白质回收率的降低.     

猪血球蛋白营养分析与水解工艺优化及制备金属螯合肽的研究

以猪血球蛋白为原料,检测其基本营养指标和氨基酸组成,以水解度为指标,研究螯合pH、水解温度和时间等单因素的影响,进一步通过响应面法优化水解制备金属螯合肽。结果表明,猪血球蛋白具有优异的营养特征,且富含多种氨基酸;采用碱性蛋白酶水解猪血球蛋白制备金属螯合肽的最佳工艺为pH值10.69、水解温度48.50 ℃、水解时间6.76 h、底物添加量8%、酶与底物比(m_酶∶m_底物)1∶250,血球蛋白粉水解度的实测值为37.83%。采用ICP-AES检测猪血球蛋白肽螯合铁、铜、锌的量分别为343.80,11.34,4.53 μg/mL。     

纤维素酶法提取玉米麸皮多糖的工艺条件优化

玉米麸皮是玉米湿法加工的主要副产物,是良好的食用、药用植物多糖来源。以未脱脂玉米麸皮为原料,将经酶法去除淀粉和蛋白质后的皮渣采用复合纤维素酶法浸提玉米麸皮多糖(ECP),通过响应面法优化确定了酶法提取最佳工艺条件为:木聚糖酶∶复合纤维素酶=1∶1,加酶量1.5%(对底物),p H 4.1,底物浓度5g/100m L,酶解时间5h,酶解温度55℃,多糖得率为27.25%。玉米麸皮多糖主要组分的相对分子量为1720、911KDa,含有一定量的蛋白质和酸性糖以及微量的阿魏酸。     

酶解条件对猪血红蛋白酶解物品质及分子质量分布的影响

我国拥有丰富的猪血资源,但因其存在色泽差、血腥味重、血红蛋白不易消化吸收等问题,并没有得到充分的利用。为有效利用猪血资源,提高猪血红蛋白消化吸收效率,研究了复合蛋白酶酶解条件对猪血红蛋白酶解物水解度、色差及分子量分布的影响。结果表明,在复合蛋白酶最适温度50℃与最适pH值8.0的条件下,酶解时间对水解度的影响最大,其次是加酶量,底物质量浓度的影响最小,猪血红蛋白较佳酶解工艺为加酶量3 500U/g,底物质量浓度125g/L,酶解时间8h,此时水解度为25.57%。未经酶解的猪血红蛋白中分子量大于3kDa组分所占比例最大,超过60%,不同酶解条件下产物中的小分子肽均较酶解前均显著增加,分子量小于1kDa组分增加至72%,大于3kDa组分不超过2%,且猪血红蛋白酶解物红度值a*显著降低。该研究结果为猪血红蛋白高效利用提供理论依据。     

血红蛋白酶法脱色技术研究

为充分改善血红蛋白的感官质量并扩大其应用范围,本实验以猪血为原料,以猪血红蛋白为研究对象,采用6种食品级商业蛋白酶Alcalase 2.4L,胰蛋白酶,Flavourzyme,AS1398中性蛋白酶,木瓜蛋白酶及胃蛋白酶在各自最适反应条件下分别酶解猪血红蛋白480min,以产物色泽为评价指标,筛选出木瓜蛋白酶为最佳水解用酶。其酶解液接近无色,透明、清澈,感官质量得到了很大提升。其酶解条件为底物浓度5%,酶与底物比100U/g,温度37℃,pH值6.5。各蛋白酶水解液的色泽由浅至深依次为:木瓜蛋白酶,Flavourzyme,AS1398中性蛋白酶,胰蛋白酶,胃蛋白酶和Alcalase 2.4L。     

Effect of l-cysteine and lactose on color stability of porcine red blood cell during freeze-drying and powder storage

The objective of this work was to establish a method to stabilize the color of porcine red blood cell (RBC). First, l-cysteine was chosen to react with hemin chloride to generate a new compound. Subsequently, l-cysteine was chosen to stabilize the color of porcine RBC along with α-lactose. The results indicated that l-cysteine along with α-lactose was effective in preventing porcine hemoglobin from oxidation and delaying porcine RBC discolor during freeze-drying and powder storage. Therefore it is a potential in the stabilizing color of RBC.     

Amino acid composition,molecular weight distribution and antioxidant activity of protein hydrolysates of soy sauce lees

The proteins of soy sauce lees (SSLP) were hydrolysed by Alcalase in the presence of ultrasound or traditional water bath to obtain hydrolysates S2–S6. The analysis of protein content indicated that enzymatic hydrolysis could significantly improve the extraction efficiency of proteins. By determination of molecular weight distribution, >10 and 5–10 KDa fractions of native SSLP (S1) decreased during hydrolysis, whilst 3–5 KDa fraction increased. Gradual increases of free, total and antioxidant amino acids were observed for S1–S4, and the differences between S4 and S6 were slight. Tyrosine was the major free amino acid of S1–S6, whilst glutamic acid had the highest amount in total amino acid composition. S2–S6 showed stronger DPPH radical scavenging activities in a dose-dependent manner than S1. All the results suggested that ultrasound treatment showed an inhibition behaviour on the enzymatic hydrolysis of SSLP.     

Properties of Hemoglobin Decolorized with a Histidine‐Specific Protease

This study investigated the application of Aspergilloglutamic peptidase (AGP) on porcine hemoglobin decolorization. AGP from fungus Aspergillus niger is identified to possess a high preference towards the histidine residues. As histidine residues in hemoglobin are known to coordinate the heme group within the globin molecule, we therefore hypothesized that incubating hemoglobin with a histidine‐specific protease would efficiently separate the non‐heme peptides from the heme‐enriched peptides with a minimum degree of hydrolysis. AGP‐decolored porcine hemoglobin hydrolysates were assessed on their functional (for example, color, emulsification, foaming, and water binding) and sensory properties. The results were compared with commercially available blood‐derived proteins (subtilisin‐decolored hemoglobin hydrolysates and plasma protein). It was observed that AGP is able to effectively decolor hemoglobin. The degree of hydrolysis (DH) increased less than 3% using AGP to achieve 90% color reduction of hemoglobin, whereas a DH increase of more than 20% is needed using subtilisin. The AGP‐decolored hemoglobin hydrolysates (AGP‐Hb) possess good emulsification, foaming, and water binding properties, which are better or comparable with the plasma protein, and much better than the subtilisin‐decolored hemoglobin hydrolysates (subtilisin‐Hb). The model canned meat with addition of AGP‐Hb showed the highest value in hardness, springiness, and chewiness from the texture analysis. Furthermore, the canned meat with AGP‐Hb was found to have a better sensory profile than the ones with addition of subtilisin‐Hb and plasma protein.     

Chitosan Coating Improves Shelf Life of Eggs

ABSTRACT: Internal and sensory quality of eggs coated with chitosan was evaluated during a 5-wk storage at 25 °C. Three chitosans with high (HMw, 1100 KDa), medium (MMw, 746 KDa), and low (LMw, 470 KDa) molecular weight were used to prepare coating solutions. Coating with LMw chitosan was more effective in preventing weight loss than with MMw and HMw chitosans. The Haugh unit and yolk index values indicated that the albumen and yolk quality of coated eggs can be preserved up to 5 wk at 25 °C, which is at least 3 wk longer than observed for the control noncoated eggs. Based on external quality, consumers could not differentiate the coated eggs from the control noncoated eggs. Overall acceptability of all coated eggs was not different from the control and commercial eggs.     

猪血中血红蛋白多肽的研究进展

猪血是生猪屠宰加工过程中的主要副产物之一,我国拥有丰富的猪血资源,但由于种种条件的限制,我国猪血利用很少,造成宝贵资源的浪费。猪血中含有丰富的蛋白质,其中血红蛋白为主要部分。因此,对血红蛋白及其水解产物多肽的研究受到了国内外众多学者的关注。本文综述了猪血中血红蛋白的分子结构和功能特性,血红蛋白多肽的制备方法、生理活性及其开发利用研究状况,并对应用前景进行了展望。     

Inhibition of hemoglobin-mediated lipid oxidation in washed fish muscle by cranberry components

Fractions enriched in phenolic acids (Fraction 1), anthocyanins (Fraction 2), flavonols (Fractions 3 and 4) and proanthocyanidins (Fractions 5 and 6) were prepared from cranberry powder using Sephadex LH-20 chromatography. Fractions 2, 3, 4, and 5 had nearly equivalent reactivity in the total phenolate assay employed per mg dry weight of each fraction while Fractions 1 and 6 were less reactive. The ability of cranberry fractions to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals as well as their inhibitory effects on hemoglobin (Hb)-mediated lipid oxidation in washed cod muscle were assessed. Addition of cranberry fractions at a level of 74 μmol quercetin equivalents per kg of washed cod muscle extended the induction time of thiobarbituric acid reactive substances (TBARS) formation in the order: Fraction 1, Fraction 3, Fraction 4 > Fraction 2 > Fraction 5 > Fraction 6. This suggests that oligomeric polyphenols (e.g., proanthocyanidins) were least effective at inhibiting Hb-mediated lipid oxidation in washed cod muscle compared to the other classes of polyphenolics in cranberry. The ability of the different cranberry fractions to scavenge DPPH radicals did not reflect their relative ability to inhibit lipid oxidation in the washed cod muscle system. Quercetin was tentatively identified as a component in cranberry that was especially effective at inhibiting Hb-mediated lipid oxidation. The ability of flavonol and proanthocyanidin-enriched fractions to inhibit Hb-mediated lipid oxidation in spite of efforts to wash away the added polyphenolics prior to Hb addition indicated these classes of polyphenolics had binding affinities for insoluble components of washed cod muscle.     

不同分子量红花籽抗氧化肽稳定性研究

目的:研究不同分子量红花籽蛋白抗氧化肽活性的稳定性。方法:以脱壳红花籽粕为实验材料,经复合酶酶解分离制备抗氧化肽,以1,1-二苯基-2-三硝基苯肼（1-diphenyl-2-trinitrophenylhydrazine,DPPH）自由基、超氧阴离子自由基（superoxide anion radical,O₂?·）及羟自由基（hydroxyl free radicals,·OH）清除能力为指标,探讨温度、pH、食品原辅料、金属离子及模拟胃肠消化等环境因素对不同分子量红花籽抗氧化肽活性的影响。结果:红花籽蛋白酶解物<3 kDa（SSPH-Ⅰ）、3~5 kDa（SSPH-Ⅱ）较分离前活性显著增加（P<0.05）,SSPH-Ⅲ（5~10 kDa）和SSPH-Ⅳ（>10 kDa）较分离前活性显著降低（P<0.05）,选取活性显著增加组分研究发现SSPH-Ⅰ抗氧化活性更加稳定,能在60~121 ℃高温、pH6~8弱酸弱碱条件下保持活性,各自由基清除率维持率达到90%以上,10 g/100 mL NaCl和葡萄糖、0.2 g/100 mL柠檬酸对SSPH-Ⅰ抗氧化活性有增强协同作用,各自由基清除率维持率增加至105%左右,10 g/100 mL的蔗糖和0.2 g/100 mL的防腐剂对活性影响不大,添加Cu2+、Zn2+和K+金属离子对SSPH-Ⅰ抗氧化稳定性显著下降（P<0.05）,其影响顺序为:Cu2+>Zn2+>K+>Mg2+>Ca2+,经模拟胃肠消化后活性较稳定,维持率为80%。结论:筛选得到<3 kDa红花籽蛋白抗氧化肽组分活性较高且稳定,为其工业化生产及应用提供理论依据。     

Effect of gamma-irradiation on the physicochemical properties of porcine and bovine blood plasma proteins

To elucidate the effect of γ-irradiation on the physicochemical properties of blood plasma proteins, bovine and porcine blood, released from a slaughterhouse, was collected and plasma proteins were prepared. Physicochemical properties of blood plasma protein powders and solutions, such as molecular weight distribution, secondary structure, solubility, and viscosity, were examined after γ-irradiation at 1, 5, 7, and 10 kGy. Oxygen radicals, such as hydroxyl and superoxide anion radicals, generated by γ-irradiation, affected the physicochemical properties of the blood plasma protein solutions, but not the plasma protein powders. Circular dichroism and sodium dodecyl sulphate–polyacrylamide gel electrophoresis studies showed that an increase of γ-irradiation decreased the ordered structure of plasma protein solutions and caused initial fragmentation of the polypeptide chains and subsequent aggregation. However, solubility and viscosity of the irradiated plasma protein powders, as well as secondary structure and molecular weight profile, were not changed significantly with radiation dose.