响应面法优化超声辅助酶解制备藏系羊胎盘肽工艺及抗氧化能力分析

以藏系羊胎盘为原料,选取水解度、肽得率和抗氧化能力为指标,在蛋白酶种类筛选、双酶组合方案确定的基础上,通过单因素试验和响应面试验优化超声波辅助复合酶解制备羊胎盘肽的工艺条件,并分析其氨基酸组成和分子质量分布。结果表明,木瓜蛋白酶和中性蛋白酶同步复合酶解效果优于其他蛋白酶组合,水解度和肽得率分别为37.24%和13.54%。响应面优化结果显示各因素对水解度影响的主次顺序为：超声时间＞超声温度＞酶解时间＞超声功率;优化后的最优工艺参数为超声时间16.5 min、超声温度25.5 ℃、超声功率438 W、酶解时间4.9 h,该最优条件下的水解度为55.22%,肽得率为18.52%,抗氧化能力（Trolox当量）为1.08 mmol/L。藏系羊胎盘肽中富含谷氨酸、甘氨酸和天冬氨酸等,分子质量低于2 000 Da的肽比例为95.02%,绝大多数为小分子低聚寡肽。可见,超声波辅助酶解制备羊胎盘肽的工艺不仅能有效改善酶解效果,还能制得具有良好抗氧化能力的小分子肽。     

牡蛎低聚肽的结构表征及体外抗氧化作用

以去壳牡蛎肉为原料,通过二步酶解法制备牡蛎低聚肽。在对其基础理化成分、氨基酸组成、相对分子质量分布分析的基础上,用扫描电镜、紫外全波长扫描和圆二色光谱对其结构进行表征,从DPPH自由基清除能力、ABTS自由基清除能力、氧自由基吸收能力（ORAC）和还原能力4个方面研究其体外抗氧化能力。结果表明：牡蛎低聚肽中蛋白质和肽含量分别为（67.57±0.53）%,（55.34±0.45）%,必需氨基酸和疏水性氨基酸含量分别为22.11%,21.43%,分子质量1 000 u以下的组分占91.16%。牡蛎低聚肽呈明显的球体颗粒状,表面有不规则褶皱和气孔,在波长267 nm处有最大吸收峰。二级结构以多种构象并存,α-螺旋、平行式β-折叠、反平行式β-折叠、β-转角和无规卷曲含量分别为（5.5±0.2）%,（3.1±0.3）%,（35.7±2.8）%,（19.9±1.1）%和（34.7±1.9）%。牡蛎低聚肽对DPPH自由基的半抑制浓度（IC50值）约为9.7 mg/mL,ORAC值为（181.75±7.26）μmol Trolox/g。当其质量浓度为5.0 mg/mL时,对ABTS自由基清除能力与0.06 mg/mL的VC相当;当其质量浓度为3.0 mg/mL时,还原能力与0.01 mg/mL的VC相当。     

超声波辅助复合酶法制备藏系绵羊胎盘肽的工艺优化

为实现羊胎盘深度开发利用,以肽得率和水解度为指标,筛选确定多种蛋白酶复合水解的最佳组合方案,并利用响应面法优化超声波辅助复合酶法水解制备羊胎盘肽的预处理参数。结果表明,响应面法优化后的超声预处理参数为超声时间18 min、温度28℃和功率420 W,超声波预处理后羊胎盘经木瓜蛋白酶和风味酶联合同步复合酶解制备羊胎盘肽的肽得率为39.89%,水解度为21.78%;该最优条件下平均肽链长度为4.59,分子量约500~600。结论:超声波辅助复合酶法制备藏系绵羊胎盘肽的工艺可行且有效。      

甲鱼肽对果蝇寿命及其抗氧化活性的影响

目的:探讨甲鱼肽（soft-shelled turtle peptide,STP）对果蝇寿命及其抗氧化活性的影响。方法:通过酶解甲鱼肉得到甲鱼肽,测定其分子量和氨基酸组成。以不同剂量（0.2%、0.4%和0.8%）的甲鱼肽饲喂果蝇,通过生存试验以及测定果蝇体内超氧化物歧化酶（superoxide dismutase,SOD）活力、过氧化氢酶（catalase,CAT）活力和丙二醛（malondialdehyde,MDA）含量,评价甲鱼肽的抗氧化作用。结果:甲鱼肽以相对分子质量介于180~500 Da的肽段为主,含有较为丰富的脯氨酸、赖氨酸和亮氨酸等。以甲鱼肽饲喂果蝇,与对照组相比,高剂量组雌、雄果蝇的平均寿命分别显著延长了10.80%（P<0.05）和14.01%（P<0.01）,平均最高寿命分别显著延长了6.35%（P<0.01）和13.22%（P<0.01）。抗氧化实验结果表明,高剂量组雌、雄果蝇的SOD活力分别显著提高了28.40%（P<0.01）和15.08%（P<0.01）,CAT活力分别显著提高了33.50%（P<0.05）和38.05%（P<0.01）,MDA含量分别显著下降了33.33%（P<0.01）和34.62%（P<0.01）。结论:甲鱼肽可以提高雌、雄果蝇体内抗氧化酶活力,缓解脂质过氧化作用,从而延长果蝇寿命,具有潜在抗衰老作用。     

河蚬抗氧化肽—锌螯合物的制备及结构表征

以河蚬肉为蛋白源,添加中性蛋白酶进行酶解,采用葡聚糖凝胶G-50分离纯化得抗氧化肽,根据螯合最佳工艺制备抗氧化肽—锌螯合物,采用茚三酮法和硫化钠法对螯合产物进行定性分析,并通过紫外、红外、荧光光谱、X射线衍射和扫描电镜对产物结构进行分析。结果表明:所得螯合产物是一种抗氧化肽—锌螯合物;抗氧化肽中的羧基、氨基及酰胺键均参与了螯合反应,反应前后肽结构发生了显著变化;根据已得图谱分析可初步预测河蚬抗氧化肽—锌螯合物的结构。     

卵白蛋白低聚肽的体外抗氧化及ACE抑制作用

以卵白蛋白低聚肽为研究对象,测定了理化成分,分子质量分布和氨基酸组成基础之上;通过DPPH·法、邻苯三酚自氧化法和水杨酸钠法测定对DPPH自由基(DPPH·)、超氧阴离子自由基(O_2~-·)和羟自由基(·OH)的清除能力,分析了其体外抗氧化作用;另外通过反向高效液相色谱法测定了对血管紧张素转化酶(angiotensin converting enzyme,ACE)的抑制作用。结果显示,分子质量低于1 000 u的卵白蛋白低聚肽占总蛋白的92.58%,并主要集中在140～500 u(占71.24%);体外抗氧化实验中,抗氧化能力与质量浓度呈现出良好的剂量关系,DPPH·、O_2~-·和·OH的半抑制浓度(IC_(50))分别为0.25、25、0.64 mg/m L;另外卵白蛋白低聚肽显示出较好的ACE抑制作用,IC_(50)值为1.5 mg/m L。结果表明,卵白蛋白低聚肽具有良好的抗氧化和ACE抑制作用,为相关抗氧化和降血压等保健类食品的开发利用提供一定的理论依据。     

鱼鳞抗氧化肽的酶法制备工艺及特性

采用碱性蛋白酶(alcalase)水解草鱼鱼鳞制备鱼鳞抗氧化肽。用单因素及正交试验考察温度、加酶量、底物水平、酶解时间等因素对酶解反应的影响,确定最适条件。以肽得率、水解度和对羟自由基清除率为考察指标,得出酶解草鱼鱼鳞的较佳工艺条件为温度55℃、底物水平10%、加酶量3%、时间5h、pH9.0;此时酶解液中鱼鳞肽得率为55.2%,羟自由基清除率为98.86%;高效液相色谱法测定鱼鳞抗氧化肽的相对分子质量排布,分子质量在1000D以下的占91.8%,等电点为2.6左右,在pH2～12的条件下溶解度均在98%以上,可广泛应用于食品中。     

芸豆肽的抗氧化活性研究

采用Sephadex-G50 凝胶柱层析分离具有强抗氧化活性的小分子芸豆肽,以VC 为对照,研究3 种肽分离组分的还原能力及对羟自由基、超氧阴离子自由基和DPPH 自由基的清除能力。结果表明：随着相对分子质量的减小和肽质量浓度的增加,其还原能力及对羟自由基、超氧阴离子自由基和DPPH 自由基的清除能力逐渐增大;小分子芸豆肽(C 组分)的还原能力低于VC;对羟自由基、超氧阴离子自由基和DPPH 自由基的半清除质量浓度分别为2.301、2.553、1.386mg/mL。该小分子肽组分的相对分子质量主要集中在100～1000,该组分疏水性氨基酸含量为36.78%,其疏水值达到364.73 kcal/mol。说明小分子肽组分的抗氧化活性与其氨基酸种类和组成、相对分子质量分布密切相关。     

核黄素结合肽的生物活性及结构表征

以松子肽Ac-QWFCT和核黄素为原料制备核黄素结合肽.利用扫描电子显微镜观察其微观特征;通过测定RAW264.7细胞活力、中性红吞噬率和NO释放率评价核黄素结合肽的免疫活性;通过测定1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基和2,2'-联氮双(3-乙基...     

食源性抗氧化肽的结构及应用研究进展

食源性抗氧化肽是指直接或间接地来源于植物或动物性蛋白质,并且具有抗氧化活性的小分子肽。近年来,食源性抗氧化肽由于其分子量小、成本低、活性强、易吸收、稳定性强,已成为一种健康、安全的抗氧化剂,也将会成为控制人体体内各种氧化过程的潜在能源。本文综述了食源性抗氧化肽的原料种类,抗氧化肽的抗氧化活性与其结构的关系,并举例说明了抗氧化肽的应用现状。     

Antioxidant and antimicrobial capacity of Chinese medicinal herb extracts in raw sheep meat

Antioxidant and antimicrobial properties of 10 Chinese medicinal herb extracts were evaluated by dipping raw sheep meat in extracts, packaging the samples in polyethylene, and refrigerating them at 4 degrees C. The optimum concentrations of Codonopsis pilosula, Platycodon grandiflorum, Artemisia capillaris, Cinnamomum cassia, Rheum palmatum, Ziziphus jujuba, Gardenia jasminoides, Santalum album, Angelica sinensis, and Bletilla striata were 0.10, 0.10, 0.25, 0.10, 0.25, 0.25, 0.25, 0.10, 0.25, and 0.25%, respectively. Analysis revealed that test ingredients were more effective in reducing lipid oxidation and microbial counts in raw sheep meat. Correlation analysis revealed a significant negative linear relationship between the inhibition of hydroxyl and lipid oxidation, and inhibition of hydroxyl was the main factor affecting lipid oxidation. A. capillaris (0.25%), C. pilosula (0.10%), and P. grandiflorum (0.10%) were identified as the most effective antioxidants. S. album (0.10%), A. capillaris (0.10%), and C. cassia (0.10%) were the most effective antimicrobials. A. capillaris (0.25%), C. pilosula (0.10%), and P. grandiflorum (0.10%) increased meat redness significantly (P < 0.05) when compared with the control samples on days 0, 3, 5, 7, 9, 10, and 11. The pH values of sheep meat treated with C. pilosula (0.10%) and A. capillaris (0.10%) were lower than those of meat treated with other extracts.     

谷朊粉钙离子螯合肽制备工艺优化及其结构表征

目的 优化谷朊粉钙离子螯合肽制备工艺并对其结构进行表征。方法 以水解度为指标,探究温度、时间、pH及加酶量等因素对谷朊粉酶解的影响;以钙离子螯合肽生成量为指标,探究不同因素对钙离子螯合肽生产量的影响;并根据结果进行Box-Behnken响应面实验优化螯合工艺。通过扫描电镜分析和红外分析对谷朊粉钙离子螯合肽进行表征。结果 最佳酶解工艺为:液料比17.5:1(mL/g)、碱性蛋白酶和风味蛋白酶添加比例为3:1、加酶量5000 U/g、酶解pH 9、酶解温度50℃、酶解时间4.0 h;优化后的最佳螯合工艺为:螯合温度38℃、肽液浓度13.5%、肽钙比5:1、螯合时间12.0 min、螯合pH 7、静置时间35 min、无水乙醇添加量为7倍体积,在此条件下钙离子螯合肽得率为50.85%。扫描电镜结果显示,螯合肽外观发生明显改变,更有利于吸收;红外光谱分析表明,钙离子可能与谷朊粉多肽上的-NH、-COOH结合。结论 本研究建立的谷朊粉酶解工艺、钙离子螯合工艺可行,可为谷朊粉深加工和产品研发提供思路与理论参考。     

Antioxidant capacity of the extracts from pulp of Osmanthus fragrans and its components

In this study, a significant inhibitory effect (EC₅₀ = 50.29 μg/ml) on the ABTS free radicals was detected in the crude ethanol extracts (CEE) of Osmanthus fragrans' pulp. Then, CEE was separated into methanol soluble fraction (MSF) and a methanol insoluble fraction (MISF), and their scavenging activities on the ABTS free radicals ascended in the order: MSF > CEE > MISF, respectively. Accordingly, the total phenolic (TP) contents of MSF, CEE and MISF were 40.53 ± 0.27, 35.37 ± 0.18 and 29.62 ± 0.23 mg/g (dry weight), respectively. Furthermore, salidroside (tyrosol 8-O-β-glucopyranoside), which showed powerful antioxidant activities in all test system was isolated from MSF. The antioxidant activity of salidroside (81.54 μg/ml) on DPPH free radicals was found to be nearly twice of that of MSF (144.78 μg/ml), and its total antioxidant activity is superior to that of the synthetic antioxidant butylated hydroxytoluene (BHT). In conclusion, our results suggested that O. fragrans' pulp, which was often considered as agricultural wastes, could be a promising source of natural antioxidants, and that MSF and salidroside may be good candidates for further development as natural antioxidants.     

棉籽多肽的制备及其自由基清除能力研究

以碱溶酸沉法提取的棉籽蛋白为原料,分别用碱性蛋白酶、中性蛋白酶、胃蛋白酶、木瓜蛋白酶对其进行水解,比较4种蛋白酶水解棉籽蛋白的水解度及棉籽多肽自由基清除能力。结果表明:中性蛋白酶水解所得棉籽多肽自由基清除能力较强。在单因素试验基础上,采用正交优化试验,得到中性蛋白酶水解制备棉籽多肽最佳条件为:水解时间5 h,水解温度45℃,pH 7.5,加酶量8 000U(100 mL 5%棉籽蛋白溶液)。在最佳条件下棉籽蛋白的水解度为11.23%,制备的棉籽多肽对超氧阴离子自由基清除率为72.48%,对羟自由基清除率为89.01%,其10倍稀释液对DPPH自由基清除率为84.67%,与0.5 mg/mL VC的自由基清除能力相当。     

三种谷物的不同乙醇浓度提取物抗氧化活性研究

以高粱、紫米、大黄米为原料,以水、60%、95%乙醇为提取介质,对提取物的总酚含量、DPPH·清除能力和总抗氧化能力进行了测定。同时,分析了三种谷物60%提取物对小鼠红细胞溶血性的影响。结果表明:高粱水提物的总酚含量为2.34mg/g(GAE/DW),分别是紫米和大黄米的4倍和12.23倍;DPPH·清除率达到43.58%,而紫米和大黄米的水提物基本无清除DPPH自由基的能力。高粱、紫米、大黄米的60%乙醇提取物的DPPH·清除能力分别为89.88%、90.16%、64.70%,高粱的总抗氧化能力为0.16mmol/g(TEAC/DW),分别是紫米和大黄米的3.99倍和24.89倍。大黄米的95%乙醇提取物总酚含量较高,达到1.38mg/g(GAE/DW),具有较强的总抗氧化能力,但DPPH·清除能力较低。高粱、紫米、大黄米在2mg/mL时对H2O2诱导的红细胞氧化溶血的抑制率分别为50.77%、16.73%、22.79%。可见,高粱具有一定的保护红细胞膜的作用。     

Antioxidant capacity of extract from edible flowers of Prunus mume in China and its active components

Prunus mume flowers are used as traditional edible and medicinal materials in China. In this study, phenolic compounds and antioxidant activity of ethanolic extract from flowers of P. mume in China were investigated for the first time. The total phenol content was estimated as gallic acid equivalents by the Folin-Ciocalteu reagent method. The antioxidant activity was measured by DPPH, ABTS⁺, and OH free radicals scavenging and ferric-reducing antioxidant power (FRAP). Three chlorogenic acid isomers, namely, 3-O-caffeoylquinic, 4-O-caffeoylquinic and 5-O-caffeoylquinic acids, were isolated and purified by preparative HPLC from the ethanolic extract and identified by UV, MS and NMR. The contents of these isolated compounds were quantified by HPLC. Results showed that 5-O-caffeoylquinic acid was of the highest level in these three isomers. The ethanolic extract demonstrated activity to some degree in all the antioxidant assays. In all tested assays, all of the isolated chlorogenic acid isomers exhibited strong antioxidant activities, which were almost the same. The results showed that chlorogenic acid isomers are the key phenolic compounds which are responsible for antioxidant activity of the ethanolic extract from Chinese P. mume flowers.     

Antioxidant capacity and total phenolic content of selected plants from Turkey

A total of 28 different plants from different regions of Çanakkale, Turkey, were investigated for their antioxidant capacity and total phenol contents to find new potential sources of natural antioxidants. The antioxidant capacity of methanolic extracts prepared from various parts of plants was evaluated by both trolox equivalent antioxidant capacity (TEAC) and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) assays, while the total phenolics were determined using the Folin‐Ciocalteu method. The TEAC values of plant extracts ranged in a large scale from 1472.36 to 17.61 μmol of trolox equivalents per g dry weight (dw), and EC₅₀ values (concentration at which 50% radical scavenging occurred) varied from 0.174 to 42.475 mg dw of plant, while the total phenol content of plant extracts ranged between 117.20 and 1.27 mg of gallic acid equivalents per g dw. There was a positive linear correlation between the TEAC and total phenols of plant materials (r = 0.916). The extracts of Hypericum perforatum, Arbutus andrachne and Paliurus spina‐christii showed higher antioxidant activities (both TEAC and DPPH assays). However, there was no clear relationship between TEAC and EC₅₀ values (r = 0.477) of plant extracts.     

黄参茎叶中酚类提取物的抗氧化活性研究

为研究黄参茎叶中酚类提取物的抗氧化活性,本文以分光光度法测定黄参茎叶中酚类提取物的总还原力、亚铁离子螯合能力、对β-胡萝卜素/亚油酸自氧化体系的抑制作用、对亚硝酸盐的清除作用,及对DPPH自由基（DPPH·）、羟基自由基（·OH）、超氧阴离子（O2-·）的清除作用,并与人工合成的抗氧化剂2,6-二叔丁基-4-甲基苯酚（BHT）和天然抗氧化剂VC进行对比。结果表明:黄参茎叶多酚具有较好的体外抗氧化性能,部分指标甚至优于阳性对照;相同浓度下黄参茎叶多酚粗提物和纯化物在不同测定体系中的清除能力有显著性差异（p<0.05）。黄参茎叶中酚类提取物可作为天然抗氧化剂进一步开发和利用。      

Antioxidant capacity of caseinophosphopeptides prepared from sodium caseinate using Alcalase

The antioxidant capacity of caseinophosphopeptide prepared at different pH from casein hydrolysate with Alcalase was evaluated using oxygen radical absorbance capacity (ORAC), reduction capacity and metal chelating activity. The yield of CPP production was dependent on the pH of supernatant from pH 4.6 clarified Alcalase hydrolysate of sodium caseinate. CPP of pH 3 had higher content of cysteine, aspartic acid, glycine, histidine, proline, tyrosine, and leucine, and less content of glutamic acid, serine, and isoleucine than those of other pH. The phosphate contents of all CPP were relatively similar, 5.0–5.4% (w/w) except at pH 3. The ORAC against peroxyl radical (ORACROO${\text{ORAC}}_{{\text{ROO}}^{\text{<glyph name="rad"></glyph>}}}$) of CPP decreased with increasing pH (3–8) in concentrations tested and was positively correlated with its reduction capacity. The ORAC against hydroxyl radical (ORACOH${\text{ORAC}}_{{\text{OH}}^{\text{<glyph name="rad"></glyph>}}}$) of CPP at 50 μg/ml increased with increasing pH to 7 and then decreased, and its metal chelating activity on ferrous ion increased linearly with increasing pH from 3 to 5, then slowly increased until pH 8, indicating that the ORACOH${\text{ORAC}}_{{\text{OH}}^{\text{<glyph name="rad"></glyph>}}}$ activity of CPP might be attributed to the scavenging activity on hydroxyl radical itself as well as the chelating activity of transition metals.     

海参多肽的抗氧化性能研究

利用海参自身的蛋白酶使其机体自溶,研究酶解液中不同分子量的海参多肽的抗氧化活性.采用超滤膜将海参酶解液分级分离,得到SPH I (＞10 kDa)、SPH II (5～10 kDa)、SPH III (3～5 kDa)、SPH IV (＜3 kDa) 4个分子量范围的海参多肽.采用化学发光体系,通过测定不同分子量海参多肽清除·OH/O2-·/H2O2的IC50,比较不同分子量海参多肽的抗氧化性,以VC做阳性对照.结果表明,4种组分对3种活性氧(ROS)的清除能力均强于VC,4种组分对3种活性氧的清除顺序分别是SPH IV＞ SPH III ＞ SPH II ＞ SPH I.说明海参多肽对3种活性氧均有良好的清除效果,并呈现量效关系.