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1.
采用响应面法优化中性蛋白酶酶解海湾扇贝(Argopecten irradias)副产物蛋白制备抗氧化酶解物工艺。以DPPH自由基清除率和还原能力为响应值,在底物浓度、温度、时间、酶与底物比4个单因素实验基础上,优化确定3个显著性因素。获得实验条件下最佳制备工艺为:底物浓度25 mg·mL^-1、温度50℃、时间3.8 h、酶与底物比1.5%。在此条件下,实验验证酶解海湾扇贝副产物蛋白制备抗氧化酶解物的DPPH清除率为81.93%(预测值为83.19%),还原能力为58.12%(预测值为58.92%),DPPH自由基清除率和还原能力的IC 50值分别为0.264 mg·mL^-1和3.136 mg·mL^-1,且表现出较强的抗氧化活性。研究表明,优化后的回归模型用于预测中性蛋白酶酶解海湾扇贝副产物蛋白制备抗氧化酶解物是可行的。 相似文献
2.
为制备抗氧化活性良好的鲢鱼鱼皮蛋白肽,采用胰蛋白酶、碱性蛋白酶、菠萝蛋白酶和木瓜蛋白酶等4种常见的商业酶对鲢鱼鱼皮进行酶解,测定酶解物的ABTS自由基清除力和Fe2+螯合力来评价其抗氧化活性,并用超滤及凝胶层析对酶解物进行分离,以期得到活性更好的酶解物分离组分。酶解后产物的抗氧化活性均有所提高,其中碱性蛋白酶酶解2 h产物活性较强。对此酶解物用截留分子量为10 k Da、5 k Da和3 k Da的中空纤维超滤膜进行超滤,得到的4个组分中,分子量越小的组分抗氧化活性越强。分子量小于3 k Da的组分经Sephadex G-15凝胶层析得到3个组分,其中分子量最大的组分活性较好,在0.51 mg/m L质量浓度下测定其ABTS自由基清除率和Fe~(2+)螯合力分别为(79.65±0.87)%和(93.40±0.20)%。该研究成果对鲢鱼鱼皮抗氧化肽的开发具有较好的指导作用。 相似文献
3.
鲢酶解物对羟自由基的清除作用 总被引：29，自引：0，他引：29
通过测定酶解物对Fenton体系产生的羟自由基的清除效果，从胰蛋白酶、木瓜蛋白酶、胃蛋白酶、枯草杆菌蛋白酶和复合蛋白酶5种酶中，筛选出木瓜蛋白酶和胰蛋白酶作为酶解鲢制备具有较高清除羟自由基活性酶解物的理想水解酶；用正交试验L9(3^4)对两种酶的水解条件进行了优化，并对最佳酶解条件下得到的酶解物进行Sephadex G-25凝胶柱分离，洗脱液分别在波长280nm处比色，测定酶解物中主要抗氧化活性肽的分子量分布。结果表明，木瓜蛋白酶在温度50℃、酶解时间15min、pH=6．5、酶质量分数1．50％、底物：水=1：2的水解条件下，酶解物对羟自由基清除效果较好，清除率为88．2％；胰蛋白酶在温度55℃、酶解时间60min、pH=8．0、酶质量分数0．25％、底物：水=1：2的水解条件下，酶解物对羟自由基清除效果较好，清除率为84．2％。木瓜蛋白酶酶解物在最大洗脱峰时有最大羟自由基清除率峰，清除率为95．1％，在最大峰处酶解物中活性肽的分子量为2．2kDa；胰蛋白酶酶解物在最大洗脱峰时也有最大羟自由基清除率峰，其清除率为89．6％，该峰处活性肽的分子量为14．2kDa。 相似文献
4.
鱼蛋白粉酶水解产物对DPPH自由基清除能力的研究 总被引：1，自引：0，他引：1
以鱼蛋白粉为原料,碱性蛋白酶(Alcalase 2.4 L)为水解用酶,研究了底物浓度、加酶量、水解p H、温度和时间对鱼蛋白粉水解产物的DPPH自由基清除率的影响。结果显示,5个因素对DPPH自由基清除率均有不同程度的影响。在单因素试验的基础上,采用响应面法对鱼蛋白粉的酶水解条件进行优化,得到的最佳水解工艺条件为:底物浓度(w/v)18.05%,加酶量(w/w)2.40%,水解p H 8.04、温度49.0℃、时间4.0 h。在此条件下水解度为9.81%,鱼蛋白粉水解液的DPPH自由基清除率为76.10%,与浓度为120μg/m L的还原型谷胱甘肽的DPPH自由基清除率相当。结果表明,鱼蛋白粉酶水解产物具有较好的清除DPPH自由基的能力。 相似文献
5.
响应面法优化末水坛紫菜蛋白酶解工艺及其酶解液抗氧化活性研究 总被引：1，自引：0，他引：1
为实现末水坛紫菜(Porphyra haitanensis)的高值化利用,研究了末水坛紫菜的蛋白酶解工艺及其酶解液的抗氧化活性。以酶解产物水解度和还原力为指标,分别采用单因素和响应面优化实验筛选出最适蛋白酶和最佳酶解工艺参数;通过测定酶解液还原力对1,1-二苯基-三硝基苯肼(DPPH)自由基、羟自由基(·OH)和超氧阴离子自由基(O_2^-·)的清除作用,研究了最高水解度下的酶解液的抗氧化性活性。结果表明,中性蛋白酶是6种蛋白酶中的最适用酶;最佳酶解条件为:底物质量浓度35 g·L-1、加酶量31 200 U·g-1、温度45℃、pH 7.6、酶解时间5 h,在此条件下坛紫菜水解度达31.37%;酶解液还原力为2.2,对DPPH、·OH和O_2^-·自由基清除率分别为56.26%、85.84%和72.73%。结果表明,中性蛋白酶可以有效水解末水坛紫菜,水解后的酶解产物具有较好的抗氧化能力和应用前景。 相似文献
6.
以印度洋鸢乌贼(Symplectoteuthis oualaniensis)胴体分离蛋白为原料,基于分子量分布和营养价值分析,优化制备抗氧化肽的酶解工艺参数。碱溶酸沉法从鸢乌贼胴体提取分离蛋白;以底物浓度、酶底比、酶解时间等3个因素为单因素实验;Box-Behnken中心法则设计响应面实验;DPPH自由基清除率结合水解度为响应指标。鸢乌贼胴体分离蛋白相对分子量分布在30~240 kDa;必需氨基酸占总氨基酸42.56%;最佳酶解工艺参数是底物浓度4%、酶底比9 U·mg-1、酶解时间4 h。5 mg·mL-1的鸢乌贼胴体分离蛋白酶解物的DPPH自由基清除率为55.60%,羟自由基清除率为53.21%,ABTS自由基清除率为40.12%。鸢乌贼胴体分离蛋白符合FAO/WHO世界卫生组织提出的理想蛋白营养价值模式,鸢乌贼胴体能制备出营养价值高和抗氧化活性强的海洋功能蛋白肽。 相似文献
7.
本研究以鳕鱼(Gadus morhua)鱼鳔为原料,选用6种不同蛋白酶对其进行酶解,通过比较酶解液的水解度与对DPPH自由基的清除能力,筛选出最佳蛋白酶为复合蛋白酶。通过单因素实验与响应面优化实验,确定最佳提取工艺：酶解时间为6 h、pH为7.21、温度为58.56℃、酶添加量为200 U/ml。研究结果显示,DPPH自由基清除率为61.1%,与理论值62.0%接近。鳕鱼鱼鳔肽体外清除自由基能力实验发现,酶解产物对羟基自由基与超氧阴离子自由基具有一定的清除能力,同时,具有较好的亚铁离子螯合能力。体外模拟胃肠消化后多肽的抗氧化活性变化,结果显示,体外模拟消化产物水解度有增加,对自由基的清除能力与亚铁离子螯合能力有一定的下降,其可能原因为,经模拟胃肠消化后多肽的结构发生一定的变化,导致了抗氧化活性的变化。 相似文献
8.
黄缘盒龟肉的酶解工艺优化及其体外抗氧化活性研究 总被引：1，自引：1，他引：0
龟类不仅营养价值高,而且具有潜在的药用价值,通过研究龟肉酶解产物的功能特性,可以科学认识其营养保健功能.以黄缘盒龟肉为原料,羟自由基清除率为指标,碱性蛋白酶为水解酶,通过正交试验L9(34)得到制备抗氧化肽的最佳酶解条件为pH值8.0、酶解温度55℃、料液比1.5∶20 g/mL、加酶量(酶/底物,E/S)3.0％、酶解时间3h,酶解液对羟自由基清除率达到82.08％.酶解产物体外抗氧化活性的结果表明,酶解产物对羟自由基、超氧阴离子自由基、DPPH自由基和过氧化氢均有较好的清除作用,还具一定的还原能力、亚铁离子螯合能力和亚油酸自氧化抑制能力.总体而言,黄缘盒龟酶解产物在不同的体外抗氧化体系中均表现出一定的抗氧化效果,具有良好的开发利用前景. 相似文献
9.
大鲵肉酶解产物的制备及其抗氧化性的研究 总被引：1，自引：0，他引：1
以大鲵肉为原料,利用Aspergillus sp.酸性蛋白酶进行酶解,研究其最佳的酶解条件以及酶解产物的抗氧化作用。结果表明,大鲵肉酶解的最佳工艺条件为Aspergillus sp.酸性蛋白酶加酶量为0.4%(质量比)及底物浓度为0.1g/mL时,酶解时间5.5h,pH 2.0,温度45℃。时间飞行质谱表明酶解产物的分子量小于2 000,苯酚硫酸法测定糖含量为2%,Folin-酚试剂法测蛋白含量为93%。大鲵酶解产物清除羟基自由基(.OH)和DPPH自由基的能力随浓度升高而增强。 相似文献
10.
《南方水产科学》2019,(6)
该研究利用中性蛋白酶、Protamex1.5L和碱性蛋白酶酶解罗非鱼(Oreochromis mossambicus)鱼皮,研究了各种酶解产物对钙离子(Ca~(2+))的结合能力,比较了酶解物结合Ca~(2+)后的抗氧化能力,并分析了结合物的傅里叶红外光谱(FT-IR)。结果显示,Protamex1.5L水解罗非鱼皮2 h的酶解产物具有最高的Ca~(2+)结合率(87.79%);3种罗非鱼皮酶解产物均存在抗氧化性,其中Protamex1.5L酶解罗非鱼皮产物的1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率、·OH自由基清除率和还原力最高分别为51.46%、17.26%和0.09;罗非鱼皮酶解物(TSH)结合Ca~(2+)后,钙离子结合物(TSH-Ca)的DPPH自由基清除率和羟基自由基清除率减弱,还原力增强;FTIR分析结果表明了Ca~(2+)与TSH中氨基氮原子和羰基氧原子发生了结合。 相似文献
11.
ABSTRACTPepsin enzyme from skipjack tuna was extracted for the production of kawakawa (Euthynnus affinis) fish protein hydrolysate. Using ultra-fractionation, Kawakawa protein hydrolysates were separated into four different fractions, including fractioned protein hydrolysate I (FPH I) (< 1 kDa), FPH-II (1–3 kDa), FPH-III (3–10 kDa), and FPH-IV (> 10 kDa). The antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, inhibition of linoleic acid oxidation, reducing power tests, and chelating activity of metal ions. Results indicated that FPH II fraction peptides had higher antioxidant activity in comparison with the other fractions, followed by FPH I. Further, the fractions were evaluated for angiotensin converting enzyme (ACE) inhibition, and IC50 value ranged from 0.45 to 1.86 mg/ml with higher activity in FPH I (IC50 0.45). Finally, the amino acid profile of different fractions was analyzed. The fractions exhibited significant amounts of hydrophobic amino acids, which could perform as hydrogen donors, frustrate the free radicals, and inhibit the ACE. The recovered pepsin from the viscera was used to produce hydrolysates with good biological activities. Peptides lower than 3 KDa had antioxidant activity as positive controls and significant ACE activity. These are very important findings that could be used to conduct further research in a preclinical study of these peptides. 相似文献
12.
金枪鱼鱼骨胶原肽的制备及抗氧化活性研究 总被引：3，自引：5，他引：3
为制备金枪鱼鱼骨胶原肽,并对其抗氧化活性进行研究,利用酶解、超滤、凝胶色谱和反相高效液相色谱制备抗氧化胶原肽,采用氨基酸序列分析仪测定其氨基酸序列,利用质谱(ESIMS)确定其分子量,采用羟自由基、DPPH自由基、ABTS自由基和超氧阴离子自由基清除实验和脂质过氧化抑制实验对胶原肽抗氧化能力进行评价。结果显示,金枪鱼鱼骨胶原蛋白经胃蛋白酶和胰蛋白酶2步酶解和分离纯化得到1个十肽(TFCH-P2),经氨基酸序列分析和质谱(ESIMS)确定其氨基酸序列为Gly-Pro-Ala-Gly-Pro-Ala-Gly-Glu-Gln-Gly(GPAGPAGQEG),分子量为839.87 u([M+H]+840.68 u)。体外抗氧化实验结果表明,GPAGPAGQEG对羟自由基(EC500.18 mg/mL)、DPPH自由基(EC500.97 mg/mL)、ABTS自由基(EC500.52 mg/mL)和超氧阴离子自由基(EC500.38 mg/mL)具有良好的清除作用;GPAGPAGQEG亦显示出良好的脂质过氧化抑制作用。研究表明,胶原肽GPAGPAGQEG抗氧化活性良好,可以用于抗氧化相关的功能食品、药物或者食品添加剂。 相似文献
13.
1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging by protein hydrolyzates
from tuna cooking juice 总被引：4，自引：0，他引：4
ABSTRACT: Protease XXIII, from Aspergillus oryzae , was used to hydrolyze tuna cooking juice at 37°C for up to 6 h. The hydrolyzate obtained at the degree of hydrolysis of 25.68% (after hydrolysis for 2.5 h) displayed the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging effect, reaching 82.19%. Six major fractions (A, B, C, D, E, and F) of this hydrolyzate were obtained by Sephadex G-25 column chromatography using a 0.05 M phosphate buffer (pH 6.5) as the mobile phase. All six fractions displayed a scavenging effect for the DPPH radical, but the scavenging effect was only obvious in two fractions (B and C). After the solid content of hydrolyzates was concentrated from one to five times, the scavenging effect of the DPPH radical increased from 17% to 75% for fraction B, and from 13% to 66% for fraction C. Seven anti-oxidative peptides were isolated from the hydrolyzates (mixture of B and C fractions) by reversed-phase HPLC. The peptide sequences comprised four to eight amino acid residues, including Val, Ser, Pro, His, Ala, Asp, Lys, Glu, Gly, or Tyr. 相似文献
14.
Schizochytrium meal protein (SMP) extracted from Schizochytrium meal was hydrolyzed by flavourzyme. Response surface methodology (RSM) was used to optimize the extraction conditions for the protein extraction yield from Schizochytrium meal. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (DRSA) was used to evaluate the antioxidant activity of hydrolysates. The orthogonal test was used to investigate the effects of three independent variables, namely protease dosage, hydrolysis time, and pH. The optimum conditions obtained were as follows: protease dosage of 7%, hydrolysis time of 1.5 h, pH of 6, under which, DRSA at the concentration of 2 mg/mL was 89.38%. Aspartic and glutamic acid constituted approximately 26.32% of the total amino acids, and glutamic acid was the most abundant amino acid of Schizochytrium meal protein hydrolysate (SMPH) by amino acid composition analysis, which may have contributed to the scavenging activity of SMPH. Moreover, SMPH was made into chewable tablets with suitable formula and high humidity stability. These findings indicate that Schizochytrium meal can be reused as a raw material for preparation of antioxidant peptides. 相似文献
15.
Penprabha Piyadhammaviboon Wasana Wongngam Soottawat Benjakul 《Journal Of Aquatic Food Product Technology》2013,22(3):265-278
Solid wastes from threadfin bream (Nemipterus spp.) surimi production composed of head and frame were hydrolyzed by various commercial proteases (Alcalase, Flavourzyme, Neutrase, Protamex, papain, and pepsin) to produce protein hydrolysates with bioactive properties. An Alcalase-hydrolyzed sample at 24.4% degree of hydrolysis (DH) displayed the highest antioxidant activity based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) assay, and potassium ferricyanide method. In addition, it showed an inhibitory activity toward angiotensin-converting enzyme (ACE) of 25.5%. Antioxidant activity of threadfin bream by-product hydrolysates increased with hydrolysis time and reached the highest DPPH activity after 6 h, while that hydrolyzed for 3 h showed the highest reducing power based on FRAP and potassium ferricyanide assays. In addition, ACE inhibitory activity was found to be at an optimum after 3 h of hydrolysis. The hydrolysates (1 mg/mL) also retarded oxidation of a linoleic acid emulsion system to a similar extent as 0.1 mg/mL 3-tert-butyl-4-hydroxy anisole (BHA), indicating a potential use in the food system. Protein hydrolysates from threadfin bream surimi by-products could be tailor-made to possess both antioxidant and ACE inhibitory activity through controlling DH of Alcalase-catalyzed reactions. 相似文献
16.
研究大眼金枪鱼头蛋白酶解物1 ku超滤组分体外的还原力、自由基清除能力及对衰老小鼠体内抗氧化能力的影响,分析1 ku超滤组分的一般成分、氨基酸组成及分子量分布,为进一步分离纯化金枪鱼头抗氧化肽提供基础。体外结果显示,1 ku超滤组分对羟基自由基、超氧阴离子和DPPH自由基的清除活性随浓度的增加而增强,IC50分别为1.38、0.73与0.93mg/mL,还原力也随浓度的增加而增大,在浓度为12.5 mg/mL时为0.763;体内结果显示,灌胃30 mg/kg的1 ku超滤组分连续42 d,D-半乳糖致衰老小鼠肝组织的超氧化物歧化酶(SOD)活性、肝组织和血清的谷胱甘肽过氧化物酶(GSH-PX)活性显著提高(P<0.05),血清丙二醛(MDA)含量显著降低(P<0.01);理化分析结果显示,1 ku超滤组分(干基计)蛋白质含量为96.40%,脂肪0.11%,灰分4.86%,疏水性氨基酸占氨基酸总量的35.8%,活性组分分子量在1 802~2 519 u和422~922 u。 相似文献
17.
Jia-Run Han Zhi-Mo Zhu Na Sun Yue Tang Cui-Ping Yu 《Journal Of Aquatic Food Product Technology》2017,26(8):993-1002
Maillard reaction products (MRPs) were produced in sea cucumber gut hydrolysates (SCGHs)-ribose system at 75°C, 85°C, and 95°C with a pH value of 7.0 or 8.0 for 12 hour. The parameters of amount of intermediate products, fluorescence intensity, browning development, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability showed a positive correlation with each other in SCGHs-ribose system. The amount of intermediate products, fluorescence intensity, and browning development followed the zero-order kinetic with activation energy from 62 to 91 kJ/mol in the Maillard reaction of SCGHs-ribose system. Moreover, 2-methyl-butanal has positive and significant correlation with DPPH radical scavenging ability of SCGHs-ribose MRPs. These results suggest that Maillard reaction improves the antioxidant activity of SCGHs, which contributes to the generation of intermediate, fluorescent, browning, and volatile products of SCGHs-ribose system. 相似文献
18.
Shabeena Yousuf Naqash 《Journal Of Aquatic Food Product Technology》2013,22(3-4):180-192
The focus of the study was to investigate the antioxidant activity of hydrolyzed muscle protein of Nemipterus japonicus and Exocoetus volitans. The trypsin protein hydrolysates of both fish showed maximum free radical scavenging potential and lipid peroxidation inhibition. Furthermore, it was purified by chromatographic methods followed by the lipid peroxidation inhibition; free radical scavenging assay was performed before and after purification. The purified peptide fractions of N. japonicus and E. volitans exhibited higher activity against polyunsaturated fatty acids (PUFA) peroxidation which was similar to natural antioxidants like α-tocopherol. Free radical scavenging potencies were measured by electron spin resonance technique (ESR). The purified peptide of E. volitans quenched free radicals (DPPH, hydroxyl, and superoxide) slightly more than N. japonicus. The amino acid composition of both fish protein hydrolysates showed variations in their ratio. The purified peptides were tested for cell cytotoxicity for Vero (kidney epithelial cells of the African Green Monkey) and Hep G2 (human hepatocellular liver carcinoma) cell lines. It was found that peptides did not show any cytotoxic effect for Vero cell lines and exerted a significant antiproliferative effect on Hep G2 cell lines. 相似文献