湛江对虾副产物酶解制备抗氧化肽

以湛江对虾加工副产物虾头为研究对象,研究虾头的酶解条件和酶解产物的抗氧化活性,选用木瓜蛋白酶和胃蛋白酶分别对虾头酶解,制备高活性的抗氧化肽。在单因素的基础上,以清除DPPH自由基为指标,研究时间、温度、加酶量、pH对酶解产物抗氧化活性的影响。结果表明,与木瓜蛋白酶相比,胃蛋白酶酶解产物的抗氧化效果更好;通过正交试验,确定胃蛋白酶酶解产物的最佳条件为温度32℃、酶解时间1 h、pH 1、加酶量为1.33×10-5kat/g(以底物计),在此条件下酶解产物的抗氧化活性最高,对DPPH自由基清除率为89.12%。     

酶解虾副产物制备抗氧化肽的研究

以虾副产物为原料,采用α-胰凝乳蛋白酶和嗜热菌蛋白酶进行酶解制备抗氧化肽。酶解液经透析、Sephadex G-15凝胶过滤层析、离子交换色谱和反向高效液相色谱分离,以1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除能力和铁离子还原能力（FRAP）为指标进行纯化,得到了高抗氧化活性组分F1。质谱分析结果表明组分F1中有三个肽,对这三个肽进行序列合成并测定其活性,发现十肽GCKVALIVVG的活性最高,其DPPH自由基清除能力按抗坏血酸当量计（AAE）为（16.10±0.02）μmol AAE/g pro,铁离子还原能力为（245.37±0.03）μmol AAE/g pro。本实验研究制备并分离纯化得到高活性的抗氧化肽,为虾副产物的生产应用提供了理论依据。      

银鲳酶解物抗氧化活性研究

选用胃蛋白酶、胰蛋白酶、碱性蛋白酶和中性蛋白酶对银鲳蛋白进行酶解以制备蛋白酶解物,以羟基自由基清除活性为指标确定银鲳最佳水解酶。结果显示,碱性蛋白酶的水解物抗氧化活性最强。实验对碱性蛋白酶水解银鲳的酶解条件(时间、温度、pH、酶添加量和固液比)进行正交实验设计,并对最佳水解条件下所获得的酶解物进行抗氧化活性测试。结果表明,银鲳蛋白碱性蛋白酶水解物对DPPH自由基和羟基自由基具有清除作用,其自由基清除效果呈现剂量依赖性,而且银鲳蛋白水解物还具有明显还原能力。所有这些体外抗氧化数据说明,银鲳蛋白水解物有明显的抗氧化效力。     

银鲳酶解物抗氧化活性研究

选用胃蛋白酶、胰蛋白酶、碱性蛋白酶和中性蛋白酶对银鲳蛋白进行酶解以制备蛋白酶解物,以羟基自由基清除活性为指标确定银鲳最佳水解酶。结果显示,碱性蛋白酶的水解物抗氧化活性最强。实验对碱性蛋白酶水解银鲳的酶解条件(时间、温度、pH、酶添加量和固液比)进行正交实验设计,并对最佳水解条件下所获得的酶解物进行抗氧化活性测试。结果表明,银鲳蛋白碱性蛋白酶水解物对DPPH自由基和羟基自由基具有清除作用,其自由基清除效果呈现剂量依赖性,而且银鲳蛋白水解物还具有明显还原能力。所有这些体外抗氧化数据说明,银鲳蛋白水解物有明显的抗氧化效力。      

蜂王浆水溶性蛋白及其酶解产物的抗氧化活性

本研究分别采用清除二苯基-2- 苦肼自由基(DPPH·),抑制亚油酸氧化和总抗氧化能力测定等方法,探讨蜂王浆水溶性蛋白及其酶解产物的抗氧化特性。结果表明,蜂王浆水溶性蛋白(WSPs)本身的抗氧化活性很弱,但酶解后抗氧化活性增强,其中胃蛋白酶和胰蛋白酶共同酶解产物(PT-WSPs)的抗氧化活性最强。对WSPs 及其酶解产物进行膜分离后发现,分子量小于3kD,尤其是小于1kD 的肽类物质其清除DPPH·、抑制亚油酸氧化及总抗氧化能力最强。     

贻贝蛋白的酶解及其酶解物的抗氧化活性研究

比较了6种不同的蛋白酶(胰蛋白酶、风味蛋白酶、中性蛋白酶、木瓜蛋白酶、复合蛋白酶、碱性蛋白酶)对贻贝粗蛋白的酶解效果,确定碱性蛋白酶为最适用酶。用此酶制备不同水解度(DH 6%、DH 11.5%、DH16%、DH 20%、DH 25%)的贻贝蛋白酶解物,考察不同DH酶解产物的抗氧化活性。试验结果表明:贻贝酶解产物具有较强的抗氧化活性,并呈一定的量效关系;当DH为25%时,贻贝酶解物对DPPH自由基、超氧自由基、羟基自由基的清除率最高,分别为77.4%、75.2%、43.4%,同时具有最强的金属螯合率(64.7%);而DH为16%时,酶解物对亚油酸的过氧化抑制作用较还原型谷胱甘肽强,达65.6%。DH对酶解产物的抗氧化活性有一定的影响,但在不同的抗氧化体系中,影响趋势不一致。     

鸡骨酶解物的抗氧化活性研究

分别选用复合风味蛋白酶、Protamex复合蛋白酶、Alcalase蛋白酶、胰酶、胰蛋白酶、AS.1398中性蛋白酶和木瓜蛋白酶水解鸡骨泥,研究鸡骨酶解物(蛋白质含量3 mg/mL)对DPPH、羟自由基(OH·)的清除能力及其还原性.试验结果表明:Protamex复合蛋白酶酶解鸡骨产物的DPPH清除率最强,即81.44%;复合风味蛋白酶酶解物OH·清除率达30.32%,还原性测定的吸光值为0.3.总之,鸡骨酶解物的DPPH清除能力强于羟自由基清除能力和还原力.在研究不同浓度和水解度对鸡骨酶解物清除DPPH活性的影响时发现,随着酶解物蛋白质质量浓度的增大,酶解物的DPPH清除率升高.复合风味蛋白酶、Protamex复合蛋白酶的鸡骨酶解物蛋白质质量浓度分别为4、6mg/mL时,其DPPH清除率分别达91.49%、97.3%.但DPPH清除率与水解度不呈正相关性,只有在特定水解度下,使抗氧化肽含量最高时,水解物才表现出最强的抗氧化能力.例如复合风味蛋白酶、AS.1398中性蛋白酶、Protamex复合蛋白酶、胰酶的最佳水解度分别为12.59%、23.78%、14.45%、20.04%.     

扇贝抗氧化肽对衰老小鼠体内抗氧化活性研究

目的研究扇贝副产物抗氧化肽对衰老小鼠体内抗氧化活性。方法采用D-半乳糖构建小鼠衰老动物模型,分别以不同性别小鼠肝脏中特征酶超氧化物歧化酶(superoxide dismutase,SOD),过氧化氢酶(catalase, CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)活性及丙二醛(alondialdehyde, MDA)含量为指标,评价扇贝抗氧化肽的体内抗氧化功能。结果与模型组比较,灌胃剂量为0.5 mL/kg的样品组能够显著提高模型小鼠(雌性和雄性)中肝脏中SOD、CAT、GSH-Px等特征酶的活性,并降低MDA含量水平。同时以脾脏指数、胸腺指数、吞噬指数为指标评价扇贝抗氧化肽对小鼠免疫能力的影响。结果显示灌胃剂量为0.50mL/kg和1.0mL/kg的扇贝抗氧化肽能够提高小鼠的脾脏指数、胸腺指数和吞噬指数。结论扇贝抗氧化肽具有良好的增强机体抗氧化功能作用。     

酶解蛋白制备抗氧化肽研究进展

从蛋白质酶解产物中制备抗氧化活性肽研究受到高度重视,对蛋白质资源开发利用具有重要意义。该文介绍近年来这一领域研究进展。     

鲶鱼骨酶解物抗氧化活性及其稳定性研究

探讨鲶鱼骨酶解物体外抗氧化活性及其抗氧化稳定性。测定不同浓度酶解物对羟自由基(.OH)、H2O2和超氧阴离子自由基(O2-)的清除效果,观察其抗氧化活性及其剂量效应关系;将酶解物溶液进行不同pH、温度、微波以及紫外光照射处理,观察其抗氧化活性的变化。结果表明:在试验浓度下,酶解物对.OH、H2O2和O2-均有较强的清除能力,其清除作用随着酶解物浓度的增加而增强,清除作用的IC50分别为6.47、15.96、12.50 mg/mL;在pH2~9范围内、80℃~121℃加热条件下,以及20 min~120 min紫外线照射和0 min~5 min微波处理时,酶解物的总体抗氧化活性变化不大。鲶鱼骨酶解物具有较强的抗氧化作用且抗氧化活性基本稳定。     

Purification and characterisation of an antioxidative peptide from enzymatic hydrolysates of duck processing by-products

Duck processing by-products were hydrolysed using eight proteases to produce an antioxidative peptide. Of the various hydrolysates produced, the pepsin extract exhibited the highest hydroxyl radical scavenging activity. The derived peptide was purified using high-performance liquid chromatography (HPLC) to identify any potent radical scavenging activity. The sequence of the antioxidative peptide obtained was identified as Asp-Val-Cys-Gly-Arg-Asp-Val-Asn-Gly-Tyr, with a molecular weight of 1096 Da. The IC₅₀ value of purified peptide for hydroxyl radical scavenging activity was 75 μg/ml as the measurement by an electron spin resonance (ESR) spectrometer. In addition, the purified peptide exhibited a protective effect on H₂O₂-induced DNA damage. These results indicate that the purified peptide possesses a potent antioxidative activity and protective effect of DNA against H₂O₂-induced oxidative damage.     

水酶法从油菜籽中提取油及水解蛋白的研究

采用水酶法工艺从双低油菜籽中提取油及水解蛋白,研究了不同细胞壁降解酶对油菜籽细胞壁的降解效果,并对蛋白酶进行了筛选.初步研究了水酶法工艺得到的菜籽水解蛋白的还原能力和清除DPPH自由基的能力.结果表明:不同细胞壁降解酶中,果胶酶c的作用效果最好.果胶酶c较佳的酶解条件为料液比1∶5(m/V)、加酶量2%、pH 3.8、50℃下酶解4 h,在此条件下油脂提取率达到92.38%.以植物蛋白酶进一步作用于果胶酶c酶解后的体系,蛋白提取率为95.72%,清油得率为88.09%,水解蛋白得率为82.50%.菜籽水解蛋白显示出很强的还原能力和清除DPPH自由基能力.     

沙棘籽酶解多肽的抗氧化性能研究

目的:研究沙棘籽三种蛋白酶水解多肽的抗氧化效果。方法:对三种蛋白酶的水解多肽进行体外清除自由基和总抗氧能力的实验及筛选,选取效果最好的一种进行体内抗氧化实验。结果:在胃蛋白酶、胰蛋白酶和木瓜蛋白酶水解沙棘籽得到的多肽中,胃蛋白酶水解的沙棘籽多肽的综合抗氧化结果最佳;动物实验表明,胃蛋白酶水解多肽灌胃的小鼠血液中GSH、CAT等抗氧化指标与阳性组相比差异显著(p<0.05),MDA指标降低效果显著(p<0.05)。结论:胃蛋白酶水解的沙棘多肽与其他两种多肽相比具有更好的抗氧化效果。      

Production of enzymatic hydrolysates with antioxidant and angiotensin-I converting enzyme inhibitory activity from pumpkin oil cake protein isolate

Protein isolate from pumpkin oil cake (PuOC PI) was hydrolysed by alcalase, flavourzyme and by sequential use of these enzymes, respectively, and the antioxidant properties and angiotensin-I converting enzyme (ACE) inhibitory activities of hydrolysates were evaluated. Under the same reaction conditions, alcalase hydrolysates showed a higher degree of hydrolysis (DH) than did flavourzyme hydrolysates. The highest DH’s by individual enzymes were 53.23 ± 0.7% and 37.17 ± 1.05%, respectively, both at 60 min. The increase of radical scavenging activity (RSA) in hydrolysates was positively correlated with the increase of DH, for both enzymes, though hydrolysates with flavourzyme showed two- or three-fold lower RSA than with alcalase. The highest bioactive potential was determined in the alcalase hydrolysate at 60 min, with RSA being 7.59 ± 0.081 mM TEAC/mg and ACE-inhibitory activity 71.05 ± 7.5% (IC₅₀ = 0.422 mg/ml). When this hydrolysate was further hydrolysed by flavourzyme, DH increased up to 69.29 ± 0.9%, but lower RSA (4.82 ± 0.21 mM TEAC/mg) and ACE-inhibitory activity (55.81 ± 6.196%) were determined in the final hydrolysate. This study suggested that the PuOC proteins could be converted into protein hydrolysates with antioxidant and ACE-inhibitory activities by enzymatic hydrolysis. Alcalase was shown as promising enzyme in further development of bioprocesses for the production of new bioactive food ingredients.     

Characterization of growth and protein contents from microalgae Navicula incerta with the investigation of antioxidant activity of enzymatic hydrolysates

Microalgae are major primary producers of organic matters in aquatic environments through their photosynthetic activities. Benthic diatom Navicula incerta is the major component of phytoplankton and also relatively easy to cultivate, used as live food source in aquaculture. The growth characteristics of N. incerta were estimated under combinations of temperature, salinity, and light; and also its composition and antioxidant activities were determined. The maximum cell density of 87×10⁵cells/mL, was reached at 20°C, 250 μmol/m²·sec, 33‰ salinity, pH 8.3, 12:12 light:dark, and F/2 medium on 2 weeks of the culture period. The antioxidant enzymatic hydrolysates efficiently quenched different free radicals: 1,1-diphenyl-2-pycryl-hydrazyl (DPPH) (pepsin IC₅₀=196.0 μg/mL), hydroxyl (α-chymotrypsin IC₅₀=102.0 μg/mL), and superoxide (neutrase IC₅₀=169.0 μg/mL). These results suggest that the enzymatic hydrolysate from N. incerta acts as a candidate against antioxidant and could be used as a potential functional food ingredient.     

Physicochemical and rheological properties of interacted protein hydrolysates derived from tuna processing by-products with sodium alginate

The physicochemical properties of tuna protein hydrolysates were enhanced by interaction with sodium alginate. The increase in emulsifying capacity and stability was from 50 to 150 m² g⁻¹ and from 36 to 49 min, respectively. The increase in foaming capacity and stability was from 100% to 140% and from 65% to 70%, respectively. The reason for the increased physicochemical properties was the reduced zeta potential level of tuna protein hydrolysates after interaction with sodium alginate. The change in internal structure of tuna protein hydrolysates after interaction with sodium alginate was determined by SEM and FTIR. The SEM results showed that a net cross-linking structure was formed from a sheet structure after the tuna protein hydrolysates interacted with sodium alginate. FTIR demonstrated that parts of the β-sheet of tuna protein hydrolysates were changed into an irregular coiled structure or α-helix after interaction with sodium alginate. In order to understand the interacted complex better, the rheological properties of interacted tuna protein hydrolysates with sodium alginate were further determined. In this study, the one-step was developed, easy-to-operate and cost-effective process that can further add value to tuna protein hydrolysates derived from tuna processing by-products.     

鸡内金酶解物制备工艺优化及抗氧化活性研究

目的：研究鸡内金的最佳酶解工艺条件,并分析其酶解物的氨基酸组成、分子量分布及抗氧化活性。方法：以鸡内金多肽得率、酶解液水解度为指标,采用正交试验优化鸡内金的酶解工艺条件;采用氨基酸分析仪、高效凝胶渗透色谱法测定鸡内金酶解物的氨基酸组成和分子量分布;并考察鸡内金酶解物对DPPH自由基和ABTS+自由基的清除能力。结果：鸡内金最佳酶解工艺条件为以中性蛋白酶和碱性蛋白酶共同酶解,酶解温度60 ℃,酶解时间9 h,酶解pH值11,蒸煮时间6 h。该条件下鸡内金多肽得率为75.68%,水解度为14.43%;鸡内金酶解物的必需氨基酸与非必需氨基酸的比值为58.56%,具有较高的营养价值;鸡内金酶解物相对分子质量＜1 000 Da所占比例为90.2%,易于消化吸收;当鸡内金酶解物质量浓度为1.5 mg/mL时,其对ABTS+自由基的清除率达到94.19%。结论：最佳酶解工艺下,鸡内金多肽得率高达75.68%,具有较好的抗氧化活性。     

虾副产物制备的钙肽结合物理化性质研究

研究由虾加工副产物经过胰蛋白酶水解得到的多肽与氯化钙反应制备的钙肽结合物的理化性质。钙肽结合物中肽含量75.03 g/100g,钙含量11.56 g/100g,水分含量6.02 g/100g。温度对钙肽结合物的溶解度影响较小,在酸性或碱性条件下钙肽结合物溶解性很好,不溶于常见的有机溶剂。钙肽结合物的紫外、红外光谱均表明钙与肽之间形成了新的化学键。