优选南极磷虾蛋白肽抗氧化活性组分

目的制备南极磷虾抗氧化肽,优选出抗氧化活性最好的南极磷虾肽成分。方法采用脱脂、酶解、超滤等手段制备南极磷虾抗氧化肽;以DPPH自由基清除率、ABTS自由基清除率、抗氧化能力指数3个抗氧化指标和超氧化物歧化酶(superoxide dismutase, SOD)、过氧化氢酶(catalase, CAT)活力2个酶活力指标为评价指标,优选出抗氧化活性最好的南极磷虾肽组分;基于G-25凝胶层析技术对抗氧化活性最好的南极磷虾肽组分进行分离纯化,进一步基于电子顺磁共振(electron paramagnetic resonance,EPR)方法测定洗脱后各峰的DPPH自由基清除率,得到抗氧化活性最好的南极磷虾抗氧化肽组分。结果通过抗氧化指标测定,截留分子量3～10 KDa的肽的DPPH自由基清除率最高,为(30.10±1.10)%,截留分子量3 KDa的南极磷虾肽的ABTS清除能力和抗氧化指数较好,则IC50值为(0.74±0.08) mg/mL、氧化自由基吸收能力(oxygen radical absorbance capacity, ORAC)值为6.39±0.21;通过酶活力指标测定,截留分子量3 KDa的肽的SOD活力和CAT活力最好,分别为(45.7±0.13)U/mg和(17.1±0.19)U/mg蛋白质。对截留分子量3KDa和3～10KDa的南极磷虾肽进行G-25分离纯化后,测定各组分的DPPH自由基清除率,可知截留分子量3～10KDa的F2-2峰清除率最好,为(51.55±1.54)%。结论基于EPR方法优选出分子量为3～10 KDa的南极磷虾肽的F2-2组分的DPPH自由基清除率最高。     

Antioxidant Activity of Oxygen Evolving Enhancer Protein 1 Purified from Capsosiphon fulvescens

This study was conducted to determine the antioxidant activity of a protein purified from Capsosiphon fulvescens. The purification steps included sodium acetate (pH 6) extraction and diethylaminoethyl‐cellulose, reversed phase Shodex C4P‐50 column chromatography. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis indicated that the molecular weight of the purified protein was 33 kDa. The N‐terminus and partial peptide amino acid sequence of this protein was identical to the sequence of oxygen evolving enhancer (OEE) 1 protein. The antioxidant activity of the OEE 1 was determined in vitro using a scavenging test with 4 types of reactive oxygen species (ROS), including the 2,2‐diphenyl‐1‐picrylhydrazyl radical, hydroxyl radical, superoxide anion, and hydrogen peroxide (H₂O₂). OEE 1 had higher H₂O₂ scavenging activity, which proved to be the result of enzymatic antioxidants rather than nonenzymatic antioxidants. In addition, OEE 1 showed less H₂O₂‐mediated ROS formation in HepG2 cells. In conclusion, this study demonstrates that OEE 1 purified from C. fulvescens is an excellent antioxidant.     

蛋清抗氧化肽分离纯化及结构鉴定

为了获得高活性、高纯度的蛋清抗氧化肽,以蛋清酶解物为原料,依次釆用超滤、离子交换色谱、凝胶色谱分离纯化抗氧化活性较强的肽段,运用基质辅助激光解吸离子化质谱解析肽链的氨基酸序列。结果表明:超滤法分离纯化EWPH所得的三个组分中,EWPH-Ⅲ(M_W<3 kDa)组分的DPPH自由基清除率最高,达到79.62%。离子交换层析分离纯化EWPH-III所得到的碱性组分B的DPPH自由基清除率最高,达到82.05%。凝胶过滤色谱分离EWPH-III-B所得到4组分中E组分的DPPH自由基清除率最高,为88.49%。高活性高纯度EWPH-III-B-E组分的相对分子质量为237.575,该二肽的氨基酸序列为丙氨酸-甲硫氨酸。     

信阳毛尖茶末多糖的分离纯化和体外抗氧化活性研究

为实现废弃茶叶资源的再利用以及探究信阳毛尖茶叶末活性多糖的体外抗氧化活性,本研究首先利用不同的乙醇终浓度沉淀优化毛尖茶粗多糖提取,然后通过DEAE-52纤维素和Sephadex-100葡聚糖凝胶分级两次纯化,并进行纯度鉴定、分子量的测定、红外光谱分析以及体外抗氧化活性的测定等。研究结果表明,90%的乙醇终浓度得率最高,为2.47%;两次分级纯化后得到XPS-5B,纯度分别为92.4%;XPS-5B符合活性植物多糖结构特征,为β-型糖苷键多糖,分子量为41208 Da;XPS-5B体外抗氧化活性随着组分浓度的增加而逐渐增强,当XPS-5B浓度为1.20 mg/mL时对DPPH自由基和羟自由基的清除率分别为89.18%、90.62%,总还原力吸光值为0.536,与未纯化的毛尖粗多糖相比,具有较强的抗氧化活性。     

HPD600型大孔树脂纯化麦麸多酚及其抗氧化活性和定性分析

目的:建立一种HPD 600型大孔树脂纯化麦麸多酚的工艺,提高麦麸多酚的利用价值。方法:采用碱水解法提取麦麸中的结合态多酚,以单因素试验优化大孔树脂的动态吸附—解吸条件,确定最佳纯化工艺;以ABTS自由基和DPPH自由基清除率反映抗氧化活性;UPLC-MS/MS完成纯化物中多酚组分的定性。结果:在最佳工艺条件下,纯化物的多酚纯度和抗氧化活性都显著提高。纯化物中主要包含6种多酚化合物,按保留时间依次为:p-羟基苯甲酸、咖啡酸、香草醛、p-香豆酸、反式阿魏酸和水杨酸。结论:建立的大孔树脂纯化工艺能有效实现对麦麸多酚的富集,所得纯化物的抗氧化能力显著提高,同时还保持了其中多酚种类的丰富性。     

Characterisation of antioxidant peptides from enzymatic hydrolysate of golden melon seeds protein

The purpose of present research was to study novel antioxidant peptides from Golden melon seeds. Alkaline protease was used to hydrolyse the Golden melon seeds protein to obtain the hydrolysed peptides. These antioxidant peptides were purified and identified by ultrafiltration, gel filtration chromatography and RP-HPLC-ESI-MS/MS. Results showed that the peptide fraction (GMSHp3) with molecular weight (MW) <3 kDa obtained by ultrafiltration had the highest antioxidant capacity. This fraction was further purified via gel filtration chromatography into six sub-fractions, among which GMSHp3-3 exhibited the highest hydroxyl radical scavenging effect. Fraction GMSHp3-3 was further purified via RP-HPLC-ESI-MS/MS and sequenced as six potential antioxidant peptides with amino acids sequences of RMSFPVMCRN, LMRVLAQLG, ALAPLVALPAA, LVGKPAPD, LPAAHKA and AHAAGYGG, among which LMRVLAQLG, LPAAHKA and AHAAGYGG possessed effective ferric reducing power. These results indicated that novel antioxidant peptides from golden melon seeds protein hydrolysates might be potential antioxidant source of functional foods or nutraceutical supplements.     

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。     

Characterization of antioxidant activity and volatile compounds of Maillard reaction products derived from different peptide fractions of peanut hydrolysate

Five peptide fractions isolated from peanut hydrolysate were purified by ultrafiltration and gel filtration chromatography. Their reaction degrees with glucose or lecithin in Maillard reaction were compared. Glucose consumption, oxygen radical absorbance capacity (ORAC) values and volatile compounds composition of peanut hydrolysate (PH) and its peptide fractions after thermal treatment and Maillard reaction were evaluated. Results revealed that the peptide fraction of 1-3 kDa showed the highest antioxidant activity. The peptides with smaller molecular weight showed a higher reaction degree to increase antioxidant activity and to produce more volatile compounds through Maillard reaction. Analysis of molecular weight distribution showed that peptide degradation and cross-linking simultaneously occurred during the Maillard reaction. In addition, lecithin addition to peptide-glucose system caused a significant increase of antioxidant activity possibly due to more nitrogen-containing compounds formed.     

裂褶菌产内切β-1,3-葡聚糖酶的分离纯化

采用不同饱和度的硫酸铵溶液对内切β-1,3-葡聚糖酶粗酶液进行分段盐析,以确定最佳盐析条件,将盐析处理后的酶液经透析浓缩、DEAE-Sephadex A-50离子交换层析、Sephadex G-75凝胶过滤层析等进一步分离纯化,浓缩纯化后的含酶组分,经过SDS-PAGE电泳分析其纯度并初步确定其分子量。结果显示:经过纯化后的内切β-1,3-葡聚糖酶的比活力由20.90U/mg提高到933.37U/mg,纯化倍数为44.7倍,酶活回收率为11.6%,电泳分析呈单一条带,分子量近似为45ku。     

罗汉果蛋白酶的分离纯化和部分性质研究

对罗汉果蛋白酶的分离纯化和部分性质进行研究.利用硫酸铵盐析、离子交换和凝胶过滤柱层析对罗汉果蛋白酶进行了分离纯化,对纯化所得的酶进行了分子量和等电点测定.结果表明,纯化酶为电泳纯,回收率为4.2％,纯化倍数为31.1,凝胶过滤层析和SDS-PAGE电泳测得酶的分子量分别为40.3 ku和39.0 ku,等电聚焦电泳测得其等电点为8.55.