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。     

玉米源活性肽对脂质过氧化的抑制作用研究

玉米肽具有良好的自由基和活性氧清除等抗氧化活性，但对其脂质过氧化抑制作用的报道较少。本文利用碱性蛋白酶、风味蛋白酶、中性蛋白酶以及模拟胃肠道消化（胰液酶+胃蛋白酶）水解制备玉米源活性肽，测定了不同蛋白酶所制备的玉米肽对ABTS自由基的清除活性，系统评价了其对亚油酸过氧化、蛋黄脂质过氧化和动物肝脏脂质过氧化的抑制作用。结果显示不同蛋白酶制备的玉米肽均具有一定的ABTS自由基清除活性以及脂质过氧化抑制作用，其中，模拟胃肠道消化所制备的玉米肽ABTS自由基清除活性最强，清除率最高达到（78.61±0.56）%；风味蛋白酶制备的玉米肽的亚油酸过氧化抑制作用最强，抑制率最高达到（72.54±1.15）%；碱性蛋白酶制备的玉米肽的卵黄脂质过氧化的抑制作用最强，抑制率最高达到（82.50±0.46）%；模拟胃肠道消化制备的玉米肽的动物肝脏脂质过氧化抑制作用最强，抑制率最高达到（61.96±0.34）%。本研究为提升玉米蛋白粉在食品领域的应用价值提供新思路，为玉米源活性肽的脂质过氧化抑制作用及其在脂质食品中的应用提供了依据。     

牦牛血抗氧化肽制备方法对比及分离纯化研究

为研究牦牛血抗氧化肽最佳制备方法,以牦牛血为原料,分别采用枯草芽孢杆菌发酵法、碱性蛋白酶解法、菌酶联合法制备3种牦牛血抗氧化肽,依次简写为BF、AP、BA。应用羟基自由基(·OH)、超氧阴离子自由基(·O_2-)清除能力,脂质过氧化抑制能力、还原力4种体外抗氧化实验,对比3种牦牛血抗氧化肽的抗氧化活性。以超滤、凝胶层析法对制备的较高活性产物进行分离。结果表明:3种抗氧化肽活性大小为BABFAP,BA有最大的·O_2-清除能力、脂质过氧化抑制能力及还原力,表明制备牦牛血抗氧化肽的最佳制备方法为菌酶联合法;超滤分离菌酶联合制备产物获得分子量5 kD的抗氧化肽活性高于分子量10 kDa、介于5~10 kDa的抗氧化肽,对·OH的IC50值为1.62 mg/mL;该组分经凝胶层析分离后得到4个组分,其最高活性组分Ⅰ对·OH的IC50值达到0.72 mg/mL。     

抗氧化胡萝卜籽肽的分离鉴定及活性表征

为从胡萝卜籽中纯化具有较高活性的抗氧化肽。采用凝胶过滤色谱和反向高效液相色谱法对胡萝卜籽蛋白酶解液进行分离纯化,以DPPH自由基清除率为指标,最终获得2个活性较强的抗氧化肽组分F3-a和F3-b。通过液质联用LC-ESI-MS/M分析得到其氨基酸序列分别为Lys-Asp-Asn-Phe-Leu-Phe(KDNFLF)和LeuPhe(LF)。此外,对2个多肽的ABTS自由基清除率、超氧阴离子自由基清除率和脂质过氧化抑制能力进行了测定,KDNFLF和LF均表现出较强的自由基清除活性。说明纯化的胡萝卜籽抗氧化肽具有良好的活性,可以作为潜在的抗氧化剂应用于食品工业及医药行业中。     

活性玉米醇溶蛋白肽的分离纯化及其还原活性研究

碱性蛋白酶水解玉米醇溶蛋白,精制后的水解产物为玉米醇溶蛋白肽。根据玉米醇溶蛋白肽对邻苯三酚自氧化的抑制活性,采用葡聚糖凝胶层析法及高效液相层析(HPLC)法分离纯化得到活性蛋白肽的几种组分,探讨了几种组分对超氧阴离子自由基的清除活性。同时,以α-生育酚为阳性对照,探讨了活性蛋白肽及其各组分的还原力。结果表明,经SephedaxG-10葡聚糖凝胶层析分离所得组分4的抗氧化活性最强,具有较高的还原力;组分4富含谷氨酸、丙氨酸、亮氨酸、组氨酸、脯氨酸和苯丙氨酸。     

玉米醇溶蛋白肽的制备及其对亚油酸自氧化的抑制作用

以乙醇从脱脂玉米蛋白粉中萃取所得的醇溶蛋白为底物,以碱性蛋白酶为催化剂,通过水解反应制备玉米醇溶蛋白肽,经精制冻干后得到肽含量为85.73%的玉米醇溶蛋白肽固体.以α-生育酚为阳性对照,研究了玉米醇溶蛋白肽及其分离纯化组分对亚油酸自氧化的抑制作用.结果表明,玉米醇溶蛋白肽及其分离组分都具有抑制亚油酸自氧化的作用,抑制效果随其质量浓度的增大而增强.经Sephedax G-15葡聚糖凝胶层析分离所得组分2抑制亚油酸自氧化的效果最好,相对分子质量主要集中在200~800 Da之间.     

五味子蜂花粉不同萃取物对小鼠肝脏脂质过氧化及DNA氧化损伤的作用

五味子蜂花粉具有强抗氧化活性,为了探究其抗氧化活性成分分布,本研究采用体积分数75%乙醇溶液对五味子蜂花粉进行浸提,分别用石油醚、乙酸乙酯和正丁醇进行萃取,测定不同萃取物的活性成分和体外抗氧化活性,研究其对小鼠肝脏脂质过氧化的抑制作用和对·OH诱导的pBR322质粒DNA氧化损伤的保护作用。结果表明,五味子蜂花粉不同萃取物抗氧化活性及其对小鼠肝脏脂质过氧化的抑制作用和对DNA氧化损伤的保护作用均存在显著差异（P＜0.05）,其顺序为：正丁醇萃取物＞乙酸乙酯萃取物＞石油醚萃取物。正丁醇萃取物的总酚含量最高,为26.11 g/100 g,总黄酮含量为27.90 g/100 g;有很强的Fe2＋络合能力、还原力和对1,1-二苯基-2-三硝基苯肼自由基的清除能力;0.2 mg/mL正丁醇萃取物对小鼠肝脏脂质过氧化的抑制率可达85%以上。本研究为进一步分离纯化五味子蜂花粉高抗氧化活性酚类物质提供依据,同时为抗氧化功能食品的开发提供参考。     

沙棘多糖清除自由基及抗脂质过氧化作用研究

目的:探讨沙棘多糖体外清除自由基及抗脂质过氧化作用。方法:采用体外抗氧化活性法测定沙棘多糖对DPPH自由基、羟自由基和超氧阴离子自由基清除能力及沙棘多糖总还原能力;制备5%小鼠肝脏匀浆,通过模拟建立体外小鼠肝匀浆自发性脂质过氧化模型、CCl₄体外诱导小鼠肝匀浆脂质过氧化模型、H₂O₂体外诱导小鼠肝匀浆脂质过氧化模型、Fe²⁺-V_C 体外诱导小鼠肝脏脂质过氧化模型,利用TBA显色法,观察沙棘多糖对脂质过氧化的抑制作用。结果:沙棘多糖对DPPH自由基、OH自由基和超氧阴离子自由基都具有一定的清除能力,其IC₅₀值分别为1.55、1.23、8.31 mg/mL,其浓度为2 mg/mL时,还原能力稍高于同浓度V_C。沙棘多糖对小鼠肝匀浆自发性脂质过氧化及CCl₄、H₂O₂、Fe²⁺-V_C所诱导的肝脏脂质过氧化均具有抑制作用,其IC₅₀值分别为1.10、1.59、9.13、1.39 mg/mL。结论:沙棘多糖具有一定的抗氧化活性及抗脂质过氧化作用。     

玉米芯功能性低聚糖分级纯化及抗氧化活性研究

探究玉米芯功能性低聚糖分级纯化前后不同组分的组成、理化性质和抗氧化活性。以玉米芯为原料,采用活性炭柱层析法对玉米芯功能性低聚糖进行分级纯化,对纯化前后组分的聚合度、相对分子质量、阿魏酸基团以及红外光谱特性进行分析,并对酶解低聚糖粗提物(EH)及其纯化组分的自由基清除能力和抑制脂质过氧化能力进行比较分析。结果表明：活性炭水洗脱低聚糖组分(WO)、醇洗脱低聚糖组分(EO)均主要由木糖和阿拉伯糖组成,其中WO含有结合态阿魏酸,为阿魏酰阿拉伯低聚木糖;EO不含结合态阿魏酸,为阿拉伯低聚木糖。EH及其纯化组分WO、EO均具有一定的清除自由基(DPPH·、·OH、ABTS⁺·)能力和抑制脂质过氧化能力,且随浓度的增加而增加,WO抗氧化活性最强,EH次之,EO抗氧化活性较弱。     

猪骨免疫活性肽的分离纯化

为了从猪骨中分离纯化出免疫活性肽,使用超滤、sephadex G-25凝胶层析、SP-sephadex C-25离子交换层析等手段对鲜猪骨的Alcalase碱性蛋白酶的酶解液进行分离纯化,采用MTT法测定各分离产物对小鼠脾淋巴细胞的增殖活性。结果显示:猪骨酶解液经超滤分离获得的分子质量小于2 ku的组分对小鼠脾淋巴细胞增殖率最高;对分子质量小于2 ku的组分采用凝胶过滤层析,对层析后活性最高的组分再进行离子交换层析,最终得到的组分质量浓度为100μg/m L时,对小鼠脾淋巴细胞的增殖率为114.30%。     

玉米蛋白抗氧化肽的制备及其在熟肉糜中的应用

采用碱性蛋白酶(Alcalase)水解玉米蛋白制备出具有抗氧化活性的玉米蛋白肽。通过测定玉米蛋白水解物对卵磷脂脂质氧化体系的抑制作用和还原能力研究其抗氧化活性,结果表明,当水解条件为底物浓度10%,加酶量(E/S)为1:100,水解时间5h时,所得水解产物抗氧化活性最强;采用凝胶层析法对5h水解物进行分离纯化,得出抗氧化最强部分的分子量范围为147～1745。将玉米蛋白肽加入到熟肉糜中,当添加量为2%时,与对照组相比可以保持肉糜鲜红的颜色,且延缓肉糜氧化。     

Purification and identification of antioxidant peptides from corn gluten meal

Corn gluten meal was hydrolyzed by alkaline protease and Flavourzyme to obtain the antioxidant peptides. The antioxidant activities of the hydrolysates or peptides were evaluated by free radical scavenging capacity (1,1-diphenyl-2-picrylhydrazyl/2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt/hydroxyl radical/superoxide radical anion), metal ion (Fe²⁺/Cu²⁺) chelating activity and lipid peroxidation inhibitory capacity. The hydrolysates were separated by ultrafiltration, and those with molecular weight <10 kDa exhibited highest antioxidant activity in all relevant assays. The hydrolysates were subsequently purified by gel filtration chromatography, and fraction F3 showed the highest antioxidant activity. Three peptides were identified from fraction F3 using LC–ESI–Q–TOF MS/MS as Leu-Pro-Phe (375.46 Da), Leu-Leu-Pro-Phe (488.64 Da) and Phe-Leu-Pro-Phe (522.64 Da). These peptides exhibited good free radical scavenging activity and lipid peroxidation inhibitory effect. Thus, corn gluten meal may be used as a potential source of antioxidant peptides for food and nutraceutical applications.     

肉味香基对人淋巴细胞的抗氧化作用

研究肉味香基的抗氧化效果,并对抗氧化成分进行分离,得到具有高抗氧化活性的组分。利用膜分离和硅胶柱对肉味香基进行分离,用人淋巴细胞的谷胱甘肽和脂肪过氧化水平评价各组分的抗氧化能力。根据相对分子质量将肉味香基分离得到3个组分,其抗氧化能力随相对分子质量的增加而增强,即:高相对分子质量组分(H-MRPs)>中相对分子质量组分(M-MRPs)>小相对分子质量组分(L-MRPs)。H-MRPs被进一步分离得到极性由弱到强的三个组分H-MA、H-MB、H-MC,其抗氧化能力从高到低为:H-MA>H-MB>H-MC。因此,肉味香基中主要抗氧化成分是高相对分子质量组分,其抗氧化能力随极性增强而减弱。     

干酪乳清酶解产物抗氧化肽的分离和纯化

干酪乳清Alcalase 2.4L酶解的最优条件,即酶解时间为2 h、pH 9.5、酶与底物比为4%、酶解温度为50 ℃。利用超滤、葡聚糖凝胶层析和三羟甲基氨基甘氨酸-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（tricine-sodiumdodecyl sulfate-polyacrylamide gelelectrophoresis,tricine SDS-PAGE）等方法从干酪乳清中提取抗氧化性蛋白肽。分别考察操作压力、料液温度、料液pH值、操作时间对超滤膜膜通量的影响。乳清酶解物超滤的最佳条件为：压力0.25 MPa、温度30 ℃、时间120 min、初始pH 9.0。分子质量4 000～6 000 D肽的水解液脂质过氧化抑制率最高,达到47.28%。利用Sephadex G-50型葡聚糖凝胶进行纯化,将分离的组分进行Tricine-SDS-PAGE分析和脂质过氧化抑制率的测定。第34管洗脱液的脂质过氧化抑制率最高,分子质量范围为4 000～4 100 D。     

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.     

卷丹乙醇提取物及其不同极性部位抗氧化活性的比较研究

对卷丹进行抗氧化活性研究,并筛选其主要的活性部位。采用体外抗氧化活性测试方法,以清除DPPH自由基和ABTS+ ·能力、还原Fe3+能力及抑制脂质过氧化能力为指标,评价卷丹乙醇总提取物及不同极性部位的抗氧化能力。卷丹及其乙酸乙酯部位和水饱和正丁醇提取部位具有较强的清除DPPH自由基和ABTS+ ·能力、还原Fe3+能力及抑制脂质过氧化能力。     

Virgin coconut oil supplemented diet increases the antioxidant status in rats

Virgin coconut oil (VCO) directly extracted from fresh coconut meat at 50 °C temperature was tested for its effect on the activities of antioxidant enzymes and lipid peroxidation levels in male Sprague–Dawley rats, compared to copra oil (CO) and groundnut oil (GO) as control. Oils were fed to rats for 45 days along with a semi-synthetic diet and after the experimental period various biochemical parameters were done. Individual fatty acid analyses of VCO and CO were done using gas chromatography. Effect of polyphenol fraction isolated from the oils was also tested for the ability to prevent in vitro microsomal lipid peroxidation induced by FeSO₄. The results showed that GO, rich in polyunsaturated fatty acids, reduced the levels of antioxidant enzymes and increased lipid peroxidation, indicated by the very high MDA and conjugate diene content in the tissues. PF fraction from VCO was found to have more inhibitory effect on microsomal lipid peroxidation compared to that from the other two oils. VCO with more unsaponifiable components viz. vitamin E and polyphenols than CO exhibited increased levels of antioxidant enzymes and prevented the peroxidation of lipids in both in vitro and in vivo conditions. These results showed that VCO is superior in antioxidant action than CO and GO. This study has proved that VCO is beneficial as an antioxidant.     

Apple favourably affects parameters of cholesterol metabolism and of anti-oxidative protection in cholesterol-fed rats

The effects of apples on lipid metabolism were studied on 40 male Wistar rats adapted to semi-purified diets containing 0.3% cholesterol. In the experimental ‘apple’ diet, a part of starch (15%) was replaced by lyophilized apple (Gala variety). In the control diet, 13% of carbohydrate was replaced by a mixture of fructose/glucose/saccharose to match the sugar supply from the apples. The lipid source was corn oil and the dietary supply of vitamin E was reduced to 1/3 of the recommended value. The rats were sampled after 21 days adaptation. The fibre supply of the apple diet was notably low (about 2%); nevertheless, there was a slight but significant cholesterol-lowering effect in plasma, as well as in liver where cholesterol esters accumulate with cholesterol diets. The lipoprotein profile was markedly altered in apple-fed rats: a reduction of cholesterol in the triglyceride rich lipoprotein (TGRLP) fraction, together with a rise in the HDL fraction; hence there was a favourable effect in a cardiovascular protection perspective. This was paralleled by effects of the apple on cholesterol apparent absorption, which was markedly depressed, whereas bile acid digestive balance was unaffected. In parallel, there was a positive effect of the apple diet on parameters of oxidative stress prevention: higher FRAP plasma levels than in controls, together with a reduced MDA excretion in urine. In conclusion, the present work indicates that a moderate supply of dessert apples elicits interesting effects on lipid and peroxidation parameters.