乳清蛋白源铜螯合肽抗氧化活性评价

对以脱盐乳清蛋白粉为原料的乳杆菌A-2代谢产物中铜离子螯合活性多肽进行分离纯化并进行其抗氧化活性的评价。采用铜离子固定化金属离子亲和层析柱分离,筛选具有铜离子螯合活性多肽,得组分F1、F2、F3,分别经Sephadex G-15凝胶过滤柱层析经进行脱盐分离,得到组分F1-1、F1-2、F2-1、F2-2、F3-1、F3-2,采用ABTS法与抗氧化指数（oxygen radical absorbance capacity, ORAC）法进行抗氧化活性测定,组分F2-1的清除ABTS阳离子自由基能力IC₅₀值为（3.068±0.15） g/L,组分F3-1的清除ABTS阳离子自由基能力IC₅₀值为（5.510±1.56） g/L;组分F2-1的相对ORAC值为（1 246.59±0.72）μmol TE/g,组分F3-1的相对ORAC值为（518.83±1.15）μmol TE/g。组分F2-1、F3-1与铜离子螯合率分别为（59±1.45）%和（6±0.32）%。结果表明,组分F2-1、F3-1具有铜螯合活性组分,具有良好的抗氧化活性。该实验为...     

具有金属螯合活性的蚕豆蛋白酶解物的制备

目的探究蚕豆蛋白酶解物的金属螯合活性,研究其金属螯合活性与其抗氧化活性的关系。方法分别采用碱性蛋白酶、中性蛋白酶、木瓜蛋白酶对蚕豆蛋白进行酶解,并测定其水解物的抗氧化活性与金属螯合活性,选用碱性蛋白酶为酶解蚕豆蛋白制取金属螯合肽的最适酶,以酶解产物的水解度、抗氧化活性及金属螯合活性为测定指标获得合适的水解条件。结果 3种蛋白酶的蚕豆蛋白酶解产物都有金属螯合活性和抗氧化活性,碱性蛋白酶为酶解蚕豆蛋白的最适酶,最适酶解时间为4 h时,得到的酶解产物金属离子螯合率为88.22%,抑制羟自由基能力为220.70 U/mg,总还原力为0.03 U/mg。结论蚕豆蛋白酶解物具有一定的金属离子螯合活性与抗氧化活性,水解度对蚕豆蛋白酶解物的金属离子螯合活性及抗氧化活性有明显的影响,蚕豆蛋白酶解物的金属螯合活性与总还原力及抑制羟自由基能力呈现显著的正相关性,相关系数分别为0.925、0.968(P0.01)。     

蚕豆蛋白酶解物分离纯化及降胆固醇活性

采用凝胶过滤色谱对经过大孔吸附树脂纯化的蚕豆蛋白酶解物进行分离纯化,以期得到高降胆固醇活性的酶解物组分。结果表明:单因素试验得到蚕豆蛋白酶解物凝胶过滤色谱最佳分离纯化工艺条件为Sep G-10、G-25葡聚糖凝胶为柱填充材料,10 mg/m L的酶解液上样量4 m L,洗脱剂为去离子水,洗脱流速1 m L/min。凝胶过滤色谱分离纯化后得到F1~F6 6个蚕豆蛋白酶解物组分;与10 mg/m L考来烯胺散阳性对照比较,F3、F6组分对3种胆酸盐(胆酸钠、甘氨胆钠酸、牛磺胆酸钠)抑制率均高于阳性对照,其中F6组分的相对抑制率最高,分别为(274.98±0.19)%、(140.22±0.20)%、(130.99±0.22)%。     

铁离子螯合亲和层析分离抗氧化活性核桃肽

采用铁离子螯合亲和层析方法分离出具有不同抗氧化活性的核桃肽,并探讨核桃肽的抗氧化活性与铁结合能力的关系。结果表明,核桃肽在酸性条件(pH 5.5)下与铁的结合能力最强,随着pH逐渐升高,结合能力下降。磷酸氢二钠对吸附到铁亲和层析柱上核桃肽的洗脱效果最好,通过阶段洗脱,得到铁结合能力逐渐增强的核桃肽组分(F1、F2和F3)。对其总还原能力和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除能力进行测定,发现其抗氧化能力为F3>F2>F1(P<0.05)。结果表明,核桃肽的铁结合能力越强,其抗氧化活性越高。     

泰和乌骨鸡活性肽的分离及其体外抗氧化作用

利用制备型HPLC 对泰和乌骨鸡活性肽进行分离,流动相为0.01mol/L 的磷酸盐溶液,采用强酸型离子交换树脂和强碱型离子交换树脂两步脱盐法对泰和乌骨鸡活性肽各分离组分进行脱盐处理。以肌肽和抗坏血酸为对照,通过清除羟自由基作用、抑制脂质过氧化作用以及对Fe2+ 和Cu2+ 螯合能力4 个体系测定泰和乌骨鸡活性肽及其分离组分的体外抗氧化能力。结果表明：泰和乌骨鸡活性肽分离所得13 个组分均具有一定的抗氧化能力,且与质量浓度呈量效关系,绝大部分分离活性肽组分比活性总肽有更强的羟自由基清除能力、脂质过氧化抑制能力以及Fe2+ 螯合能力。     

琼脂糖-Zn（II）亲和层析分离纯化罗非鱼水解多肽初探

以广州大学生化课题组自制琼脂糖微球为载体、环氧氯丙烷(ECH)为活化剂、亚氨基二乙酸(IDA)为螯合配基、Zn(Ⅱ)为螯合金属制备琼脂糖-Zn(Ⅱ)亲和层析介质。最佳活化工艺:DMSO 6 mL、ECH 10 mL、活化温度35℃、活化时间3.0 h;最佳螯合工艺:IDA 0.9 g、反应时间4.0 h、ZnSO_4浓度0.25 mol/L,Zn~(2+)螯合量达到最大值。通过用0.05 mol/L EDTA缓冲液洗脱,有效地分离纯化罗非鱼水解多肽,得到适合与Zn(Ⅱ)螯合的目标多肽组分,并制备出多肽-Zn(Ⅱ)配合物。     

菜籽多肽体外和细胞内抗氧化性评价及氨基酸分析

本研究以Alcalase 2.4L酶解“双低”油菜籽宁杂19号得到的菜籽蛋白酶解物（rapeseed protein hydrolysate,RPH）为原料。通过超滤和凝胶色谱分离其活性肽组分,对酶解液及各分离组分进行抗氧化活性研究,并对抗氧化能力最高的组分进行氨基酸分析。结果表明：通过超滤将RPH分离成4 组不同分子质量范围的多肽组分,其中RPH-P4（分子质量＜3 kD）具有最高的抗氧化活性;RPH-P4经Sephadex G-15凝胶柱分离后得到3 个洗脱峰,收集并测定其活性,研究发现RPH-P4-S3抗氧化能力远高于其他2 个组分（P＜0.05）,氧自由基吸收能力（oxygenradical absorbance capacity,ORAC）、细胞抗氧化活性（cellular antioxidant activity,CAA）、细胞内CAA实验的EC50值分别为（1 951.90±20.35） μmol TE/g、（143.38±4.11） μmol QE/g、（45.63±3.67） μg/mL;对RPH-P4-S3进行了氨基酸分析,发现其抗氧化性氨基酸含量达到72.90%,必需氨基酸含量为53.52%。研究认为,菜籽蛋白Alcalase 2.4L酶解物分离组分RPH-P4-S3具有较高的抗氧化活性及营养价值,可以作为功能性成分用于抗氧化相关的功能食品和保健品的开发。     

榨前分离苹果皮多糖的鉴定及抗氧化活性研究

以榨前分离苹果皮为原料,采用超声波辅助法提取多糖,通过改变洗脱液的极性对多糖进行分离纯化,并对各组分的抗氧化活性进行分析。结果表明:多糖样品经过DE-52纤维素柱梯度洗脱得到三个含量较大的洗脱峰,分别为水、0.3mol/L NaCl和0.9mol/L NaCl洗脱组分。采用Sephadex G-200凝胶柱对洗脱组分进一步纯化,得到APPSH2O、APPS-0.3mol/L和APPS-0.9mol/L三种不同组分的多糖纯品。抗氧化实验表明:三种不同组分多糖均有一定的还原力、清除·OH、DPPH·,O-2·的能力,但其抗氧化能力均小于粗多糖和V C。     

大米蛋白体外消化产物抗氧化活性的研究

为了研究大米清蛋白、球蛋白、醇溶蛋白和谷蛋白体外消化产物的抗氧化活性,用Osborne法从大米粉中提取大米四种蛋白。采用胃-胰蛋白酶分步酶解,选择DPPH·,ABTS+·,·OH自由基清除能力和Fe2+金属离子螯合能力四个指标对消化产物的抗氧化活性进行综合评价。与市售的大豆肽比较,大米四种蛋白经模拟体外消化后具有良好的抗氧化活性。其中醇溶蛋白的消化产物抗氧化活性最好,DPPH·,ABTS+·,·OH自由基清除能力和Fe2+金属离子螯合能力的半抑制浓度分别为30.88 mg/m L、27.61 mg/m L、5.43mg/m L、0.18 mg/m L。大米四种蛋白酶解产物的Fe2+螯合能力半抑制浓度均小于1 mg/m L,具有良好的Fe2+螯合能力。研究结果表明大米四种蛋白的体外消化酶解物具有不同大小的抗氧化活性,分子量集中在181~1000 Da范围内,是一种易于被人体吸收的抗氧化肽。     

酪蛋白酶解物中抗菌肽的分离及其分子量的测定

使用离子交换层析分离纯化酪蛋白酶解物中的抗菌肽并测定其分子量。选用Q-Sepharose Fast Flow为分离介质,对分离条件进行了研究,并对各洗脱组分进行抑菌活性测定,确定了适宜分离条件下酪蛋白抗菌肽的洗脱体积,同时测定了酪蛋白抗菌肽的分子量。结果表明,离子交换层析对酪蛋白酶解物中的抗菌肽的最佳梯度洗脱条件为:流动相A:pH10.5的0.05mol/L乙醇胺盐酸缓冲液;流动相Β:含1mol/L氯化钠的A相溶液,pH10.5;洗脱程序:7%B,1.55CV;13%B,1.40CV;25%B,1.25CV;100%B,1.55CV,洗脱流速:0.5mL/min,检测波长:280nm,酪蛋白抗菌肽的洗脱体积为76.57mL和89.48mL。酪蛋白抗菌肽的平均分子量为3137u。     

Antioxidant properties and protein compositions of porcine haemoglobin hydrolysates

Porcine haemoglobin hydrolysates were prepared through hydrolysis by Alcalase followed by Flavourzyme, and their protein compositions were analyzed using Sephadex G-50 gel filtration chromatography. The antioxidant activities, including reducing power, ferrous ion chelating ability, and DPPH radical scavenging activity, of the hydrolysates were evaluated. The results showed that the hydrolysates of haemoglobin exhibited low reducing powers, but high ferrous ion chelating abilities and DPPH radical scavenging activities. The hydrolysate, obtained through hydrolysis by 2% Alcalase for 4 h and followed by 1% Flavourzyme for 6 h, had the highest ferrous ion chelating ability of 63.54% at a concentration of 5.0 mg/mL. The hydrolysate, obtained through hydrolysis by 2% Alcalase for 4 hrs, had the highest DPPH radical scavenging activity of 41.94% at a concentration of 5.0 mg/mL. According to the results of protein composition analysis, we divided the hydrolysates into three groups, including high molecular weight (MW) group (Group I), medium MW group (Group II), and low MW group (Group III). The reducing power and ferrous ion chelating ability of the hydrolysates were significantly and positively correlated to the relative amount of Group I, and negatively correlated to the relative amount of Group III. This study revealed that the antioxidant activities of porcine haemoglobin hydrolysates were dependent on their protein compositions. The high MW protein fraction (Group I) was responsible for the high reducing power and ferrous ion chelating ability of the hydrolysate.     

Study on Antioxidant Activity and Amino Acid Analysis of Rapeseed Protein Hydrolysates

Alkaline extraction followed by acid precipitation were employed to extract rapeseed protein and, Alcalase 2.4 L was used to obtain rapeseed protein hydrolysates. Three groups of rapeseed protein hydrolysates were obtained by purifying with membrane ultrafiltration and a Sephacryl S-100HR gel column. The antioxidant activities were then determined. Group 3 had the best antioxidant activities according to the oxygen radical absorbance capacity, peroxyl radical-scavenging capacity, and cellular antioxidant activity assays, with the following antioxidant values: an oxygen radical absorbance capacity value of 1610 ± 113 µmol TE/(g sample), a peroxyl radical-scavenging capacity value of 622 ± 30 mg VC/(100 g sample), and a cellular antioxidant activity value of 25 ± 2 µmol QE/(g sample) and corresponding EC₅₀ value of 58 ± 3 µg/mL. Six peaks of group 3 were collected and well separated by reversed phase–high-performance liquid chromatography. Peak 5 were identified to exhibit a higher antioxidant activity, the amino acid sequence of which was found to be Trp-Ile (Leu)-Tyr, as determined by liquid chromatography–mass spectrometry.     

罗非鱼肌浆蛋白源抗氧化肽的制备、分离纯化及其体外抗氧化活性

采用木瓜蛋白酶对罗非鱼肌肉蛋白组分（肌浆蛋白、肌原纤维蛋白、基质蛋白）进行酶解,研究罗非鱼不同蛋白组分酶解产物的抗氧化活性,并通过超滤、凝胶过滤色谱、反相高效液相色谱对高活性肌浆蛋白组分抗氧化肽进行分离纯化,同时对纯化后的抗氧化肽氨基酸组成予以分析。结果表明：罗非鱼肌浆蛋白酶解物（tilapia sarcoplasmic protein hydrolysates,TSPH）抗氧化活性高于肌质与肌原纤维蛋白酶解物,当肌浆蛋白酶解4 h时,TSPH（6 mg/mL）对1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基、羟自由基清除率及其还原力均达到最高,分别为84.61%、50.01%和0.629;经超滤分离后,其分子质量小于5 kDa的组分具有较高的抗氧化活性;该组分经凝胶色谱分离后,富集得到E1～E4共4 个组分,其中E3组分（0.8～0.3 kDa）的抗氧化活性最高, 3 mg/mL E3组分对DPPH自由基清除率达83.61%,还原力为0.953;E3组分再经反相高效液相色谱分离纯化后,富集得到F1～F9共9 个组分,其中F4组分的抗氧化活性最高,其DPPH自由基的半清除质量浓度为0.78 mg/mL,且经氨基酸组成分析发现色氨酸（Trp）、甘氨酸（Gly）、组氨酸（His）为其含量较高的氨基酸,相对含量分别高达31.20%、27.25%和8.97%。     

Evaluation of Antioxidant Activities of Zein Protein Fractions

Zein protein was extracted from the by‐product corn gluten meal. The obtained zein protein was 1st hydrolyzed by 4 different proteases. The antioxidant activities of the hydrolysates or peptides were evaluated by free radical scavenging activity, metal ion chelating activity, and lipid peroxidation inhibitory capacity. Among hydrolysates produced, alkaline protease hydrolysates exhibited the highest antioxidant activity. A regression model was established by uniform design to optimize the alkaline protease hydrolysis conditions. The hydrolysates with molecular weight < 3 kDa obtained from ultrafiltration showed the highest antioxidant activities in all relevant assays. The hydrolysates with molecular weight <3 kDa were subsequently purified by gel filtration chromatography, and fraction F3 exhibited the highest antioxidant activities. Two peptides were identified from fraction F3 using LC‐ESI‐Q‐TOF MS/MS as Pro‐Phe (263.13 Da) and Leu‐Pro‐Phe (375.46 Da). These peptides exhibited good free radical scavenging activity and lipid peroxidation inhibitory effect. The results clearly indicated that zein protein fractions are good sources for the development of natural antioxidants for the food industry.     

Antioxidant and metal chelating activities of Phaseolus vulgaris L. var. Jamapa protein isolates, phaseolin and lectin hydrolysates

A search in a database of potential bioactive short sequences in food proteins reveals that bioactive peptides with a variety of beneficial effects for cardiovascular health are present in the sequence of common bean proteins, including bioactive sequences with antioxidant properties. A protein isolate, the storage protein phaseolin and a lectin extract from Phaseolus vulgaris L. var. Jamapa, were hydrolyzed by treatment with pepsin and pancreatin in order to investigate the possible release of peptides with antioxidant and metal chelating properties. Antioxidant activity was determined in Caco-2 cells exposed to a free radical generator, and iron and copper chelating activities were determined using colorimetric methods. The highest antioxidant activity, 71% inhibition, was found in the hydrolyzed protein isolate. Copper and iron chelating activities were highest in the lectin and phaseolin hydrolysates, 53% and 81%, respectively. Thus, experimental data indicates, as suggested by the database search, that antioxidant peptides are abundant in pepsin-pancreatin hydrolysates, which may represent a valuable health-promoting property in common bean.     

菜籽蛋白水解物体外和细胞内抗氧化性评价及氨基酸分析研究

以“双低”油菜籽秦优7号为原料,碱溶酸沉法获得菜籽分离蛋白后进一步用Alcalase2.4L酶解得到菜籽蛋白水解物（rapeseed protein hydrolysates,RPHs）;采用聚丙烯酰胺葡聚糖凝胶柱（Sephacryl S-100HR）对RPHs进行分离纯化,得到3 个组分;采用抗氧化能力指数（oxygen radical absortion capacity,ORAC）方法以及细胞抗氧化活性（cellular antioxidant capacity,CAA）的方法分析筛选得到抗氧化性最好的组分3。结果表明组分3的ORAC值为（1610.38±112.51）μmol TE/g;CAA值为（124.66±2.18）μmol QE/g,EC50值为（57.84±3.38）μg/mL;在此基础之上,对组分3进行氨基酸分析以及电泳分析,表明其抗氧化性与分子质量分布及氨基酸组成可能存在一定关系。     

Antioxidant and metal chelating activities of peptide fractions from phaseolin and bean protein hydrolysates

Bean protein isolate and phaseolin were hydrolysed using pepsin and pancreatin, and the resulting hydrolysates were filtered through a 1kDa cut-off membrane and fractionated by size exclusion chromatography. Three fractions corresponding to MW 0.7-1.0kDa, 0.43-0.7kDa and <0.43kDa (A1, A2, and A3 for protein isolate fractions, and B1, B2, and B3 for phaseolin fractions) were assayed for antioxidant and metal chelating activity and they were also subjected to amino acid and SDS-PAGE analysis. Fractions A1 and B1 had the highest copper chelating activity (78% and 82%, respectively), while iron chelating activity was the highest in fractions A1 and B3 (36% and 16%, respectively). Fractions A2 and B3 had the highest antioxidant activity as determined by inhibition of reducing power and β-carotene bleaching, while the highest ABTS radical scavenging activity was found in A3 and B3. Thus, fractions coming from the isolate and phaseolin had similar activities except for iron chelation, suggesting that phaseolin is the major contributor to the antioxidant and copper chelating activities of the hydrolysed protein isolate.