地锦草有效成分的提取及其生物活性的研究

采用乙醇浸提法来提取地锦草中的有效成分,并对其生物活性进行研究。探讨了地锦草总黄酮的最佳提取工艺;通过体外抗氧化实验,对地锦草提取物的抗氧化活性进行研究;同时考察了提取物对大肠杆菌、枯草芽孢杆菌、青霉、曲霉的抗菌性能,并对地锦草提取物对α-淀粉酶的抑制活性进行研究。实验结果表明:地锦草总黄酮最佳提取工艺为乙醇体积分数为85%,料液比为1∶15,提取次数为3次;体外抗氧化实验表明,地锦草具有抗氧化活性,且在浓度为8 mg/mL最为明显;抗菌性能实验表明,地锦草具有抗菌性能,且对青霉、曲霉的抗菌效果比较显著;α-淀粉酶的抑制活性的实验表明,地锦草提取物对α-淀粉酶的活性有抑制能力,在pH 6.8、浓度为4.8 mg/mL、温度为70℃时,其对α-淀粉酶的抑制效果达到最大。     

两种含氮类大豆苷元衍生物对人体红细胞的抗溶血作用

合成了2种含氮类大豆苷元衍生物,4,7-二（（甘氨酸钠）羰基）甲氧基异黄酮（L1）和4'',7-二（肼羰基）甲氧基异黄酮（L2）并用元素分析、红外光谱和核磁共振氢谱对其进行了表征。研究了大豆苷元衍生物的抗氧化性能,评价了其对自由基、超氧阴离子自由基的吸附能力,以及对人体血红细胞抗氧化损伤的保护作用。实验结果表明,大豆苷元衍生物在生理pH条件下的抗氧化活性优于维生素C,特别是在清除羟自由基和抑制人血红细胞溶血方面,大豆苷元衍生物抗氧化能力表现更为突出。大豆苷元衍生物的羟自由基清除活性IC₅₀值是维生素C的104倍。     

核桃蛋白酶解物的制备及抗氧化活性

采用碱性蛋白酶对核桃蛋白进行酶解, 检测了所得酶解物的抗氧化能力, 包括对1,1-二苯基-2-三硝基苯肼(DPPH)和羟基自由基(·OH)的清除能力; 利用Sephadex G-25 凝胶层析柱和反相柱对核桃蛋白酶解物进行分离纯化; 采用液相色谱-质谱(LC-ESI-Q-TOF)联用方法测得抗氧化能力最强的多肽的序列为Ala-Gly-Gly-Ala, 其还原力和还原型谷胱甘肽相当.     

虎耳草活性成分的提取及功效评价

对虎耳草全草中总黄酮物质进行有效提取,并将蒸发浓缩后的虎耳草提取液分别进行了体外抗氧化、抗炎、抑菌性能以及美白功效等研究。结果表明,虎耳草提取液在较高浓度时的抗氧化功效与维生素C相当;具有良好的抑制炎症性能,其乙酸乙酯萃取物消炎性能更优;虎耳草提取物能够抑制常见的普通细菌的生长,其正丁醇萃取物抑菌性能最佳;在美白功效测试实验中提取液浓度为4000μg/mL时对酪氨酸酶活性的抑制率为79.25%。     

乙烯基桥联的8-羟基喹啉衍生物的合成、抗氧化活性及促进鼠骨髓间质干细胞增殖的研究

设计合成了2-乙烯基取代的8-羟基喹啉衍生物(6), 用IR, UV-Vis, 1H NMR, MS和元素分析确认其结构; 利用二苯代苦味酰基自由基(DPPH·)法和噻唑蓝比色(MTT)法分别测定了目标产物的抗氧化活性及调控鼠骨髓间质干细胞(MSCs)增殖的作用. 结果表明, 目标产物有较强的抗氧化活性, 随着产物浓度的增加, 其抗氧化活性先增强后减弱; 在低浓度时对鼠骨髓间质干细胞有促进作用. 研究发现, 化合物6抗氧化活性的提高与其含量相关.     

连翘不同部位生物活性比较研究

对青翘、老翘、连翘根和连翘茎进行指纹图谱分析,总酚和总黄酮含量测定以及抗氧化和抑菌活性评价,并探讨生物活性的化学物质基础。采用高效液相进行指纹图谱分析;利用Folin-Ciocalteu和NaNO_2-Al(NO_3)_3-NaOH比色法测定总酚和总黄酮含量;分别采用DPPH·法和ORAC法评价抗氧化活性;以最小抑菌浓度(MIC)为评价指标评价抑菌活性。青翘中的总酚和总黄酮含量较高,且其抗氧化和抑菌能力较强;老翘、根、茎均对金黄色葡萄球菌具有抑制作用和具有较好的抗氧化能力,其中,连翘茎表现出对金黄色葡萄球菌较强的抑制能力。除青翘、老翘具有药用价值外,连翘根和连翘茎也具有潜在的利用价值。     

花青素类物质对pH响应的紫外光谱学和理论研究

本文以氯化矢车菊素和氯化飞燕草色素两种常见花青素为研究对象,检测了它们在不同pH水环境下的紫外-可见吸收光谱,通过研究吸收光谱峰位置的变化,对pH变化过程中物质结构的变化进行了分析;同时通过理论计算对两种物质的结构进行了优化,根据相关数据探讨了该类花青素的电子云分布及其对抗氧化活性的影响。结果表明,随着酸碱度的变化,氯化矢车菊素可能有四种稳定结构的中间态,氯化飞燕草色素可能只有三种,说明母核上羟基取代基的数量和位置对花青素类化合物的活性和稳定性有较大影响,这也会影响其抗氧化活性。理论计算得到的轨道能级数据说明氯化飞燕草色素可能有更好的抗氧化活性;羟基的电荷分布则分析了花青素不同位点的羟基与自由基结合的能力,这也表明氯化飞燕草色素可能有更好的抗氧化活性。     

新型Trolox-壳聚糖纳米颗粒的制备及抗氧化效应

氧化应激是很多慢性疾病的重要病因之一,如癌症、动脉粥样硬化和神经退行性疾病等。抗氧化剂对于这些疾病的抵御作用吸引人们对其进行了广泛的研究。然而,目前抗氧化剂在抗氧化应激方面的应用研究,仍存在两大问题亟需解决：一、水溶性差;二、生物利用效率低。而壳聚糖纳米颗粒具有可生物降解性、生物相容性好等优点,目前被作为一种载体广泛用于输送各种生物大分子,如蛋白多肽、DNA和siRNA等,同时也为提高非水溶性物质的生物利用效率提供了新的契机。作者所在实验室在前期工作中还曾发现以纳米金为核的Trolox分子可提高其抗氧化反应的效率。在本文我们制备出小尺寸的水溶性Trolox-壳聚糖纳米颗粒(20-40nm),细胞抗氧化对比实验进一步证实其抗氧化效率明显高于Trolox单体化合物。     

呋喃甲酰壳聚糖的制备、表征及其抗氧化活性

以壳聚糖与呋喃甲酰氯反应得到呋喃甲酰壳聚糖。通过FT-IR、1 H-NMR、X射线衍射、热重分析、溶解度实验、元素分析、抗氧化活性测试等手段对产物进行了结构和性能表征。结果表明：产物为目标产物且热稳定性好于壳聚糖;在水中的溶解性能良好,溶解度为0.04g/mL;取代度为0.69;当对羟基自由基的清除率达到50%时,呋喃甲酰壳聚糖的质量浓度为1.1mg/mL,其还原能力随质量浓度增加而增强,抗氧化活性优于壳聚糖。     

肉苁蓉多糖锌的制备、表征及抗氧化活性

采用超声酶法辅助提取肉苁蓉粗多糖,并用季铵盐沉淀法对肉苁蓉粗多糖进行精制,获得肉苁蓉精制多糖(UCEP1);以UCEP1和硫酸锌为原料,制备了肉苁蓉多糖锌配合物(UCEP1-Zn),对其结构进行了表征,并研究其抗氧化活性.结果表明,肉苁蓉粗多糖收率及含量分别为22.31%和48.50%,UCEP1中多糖含量为70.13%;UCEP1-Zn中-OH和-COO~-和Zn~(2+)发生了配位,其中锌含量为7.57%,且多糖链构象及多糖分子表面形貌发生变化;UCEP1-Zn的抗氧化能力显著高于UCEP1,且对ABTS~+自由基清除活性最好.该研究可为肉苁蓉多糖、新型锌营养素补充剂以及锌功能配合物的研究和利用提供参考.     

Preparation and Antioxidative Effects of Soybean Peptides*

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A Study on Anti-oxidative Activity of Soybean Peptides with Linoleic Acid Peroxidation Systems

Introduction Freeradicalsproducedbyperoxidationcancause cellulardamageandforfeitureofavarietyofphysiologi calfunctions[1,2].Inrecentyears,ithasbeenreported thatsomenon enzymicsmallmolecularcompounds,suchasVitaminC(VC)andVitaminE(VE),glutathi one(GSH),SODm…     

Preparation,Antioxidative Effects of Soybean Peptides

In this work an alkaline protease, alcalase, was used to hydrolyze the separated soybean protein at 50℃, pH 8.0. The dependence of hydrolysis time on hydrolysis degree, relative antioxidative activity was examined. The antioxidative activity of the hydrolysate reached the maximum after the hydrolysis for two hours, then decreased. The hydrolysate hydrolyzed for two hours was isolated on a Sephadex G-25 column. The range of molecular weight for the collected fractions was 100-1 300. A fur ther purification of the isolated antioxidative peptides was conducted on a CM-Cellulose column. Two active peaks were observed. One of them was taken to investigate the antioxidative properties for in vitro model systems. It was found that the autoxidation rate of pyrogallol under the alkaline condition was decreased by, at least, 10%, indicating that some free radicals in the tested system were removed. In the experiment of the antihemolysis of erythrocyte, it was observed that the hemolysis degree of erythrocyte caused by hydroxyl free radicals was decreased obviously. This result indicates the protective effect of the antioxidative peptides on the cell membrane damage of erythrocyte. Moreover, it was also found that the membrane fluidity could be depressed after a solution of the antioxidative peptides was mixed with a liposome solution, indicating that the antioxidative peptides could fur ther inhibit the membrane damage caused by lipid peroxidation when the small molecules of antioxidative peptides were embedded in liposome.     

梅花鹿茸多肽的化学结构及生物活性

在马鹿茸活性多肽结构与功能研究基础上, 从新鲜梅花鹿茸中分离纯化了活性单体多肽, 确定了其化学结构, 并与马鹿茸多肽进行结构与活性比较. 利用离子交换层析、 凝胶过滤层析及反相高效液相色谱层析等生物化学技术, 从梅花鹿茸中分离得到1个新多肽, SDS-PAGE电泳显示为一条带, HPLC图谱为单一峰, MALDI-TOF MS给出该多肽的精确分子量为3263.4, 其等电点pI=8.15. 一级结构研究表明, 该多肽是由32个氨基酸残基组成的直链多肽, 不含半胱氨酸, 富含缬氨酸、 赖氨酸、 亮氨酸和甘氨酸, 氨基酸序列为VLSATDKTNVLAAWGKVGGNAPAFGAEALERM. 生物活性检测结果表明, 该多肽可促进原代培养的表皮细胞和软骨细胞增殖, 也能刺激NIH3T3成纤维细胞株的分裂. 梅花鹿茸多肽与马鹿茸多肽在结构上均为32个氨基酸残基组成的直链多肽, 但第5, 8, 11和30位氨基酸残基不同. 2种多肽结构上的变化并未影响其促细胞增殖生物活性.     

Preparation and Anti-oxidative Effects of Corn Peptides

IntroductionBioactive peptides include the natural peptidesfrom organism itself and the active peptide fromhydrolysate of protein. In the pastfew years,someresearchers have found that small peptides derivedfrom the hydrolysates of food proteins play animportant role in regulating autonomic nervoussystem,activating the cellular immunity function,ameliorating the cardiovascular function,antioxidizing and antiaging,etc.[1— 7] . Smallpeptides prepared from food proteins becomenitrogenous source f…     

血管紧张素转化酶活性抑制剂──丝素肽的分离、纯化和结构鉴定

 可溶性丝素粉末经碱性蛋白酶Alcalase水解后 ,其酶解产物对血管紧张素转化酶 (ACE)的活性有很强的抑制作用。采用凝胶过滤色谱SephadexG 15和反相高效液相色谱 (RP HPLC)对水解度为 2 0 %的酶解产物进行分离纯化 ,利用质谱鉴定其中一种ACE抑制剂是肽 ,其结构为Gly Tyr。     

Identification and isolation of human insulin A and B chains by high-performance liquid chromatography

A method for the isolation, identification and quantification of human insulin A and B chains by high-performance liquid chromatography (HPLC) is described. These chains were isolated from a peptide mixture produced by E. coli with modified genes obtained by genetic engineering. The method is based on the use of hydrophilic reagents, forming ion pairs in a reversed-phase column. Because some undesirable effects resulting from the use of phosphoric acid were observed, especially with the B chain, a new HPLC method was developed for each of the two human insulin chains. The use of trifluoroacetic acid as a counter ion for the A chain and of formic acid for the B chain led to the rapid isolation and purification of each chain by HPLC. The advantage of this method is that it provides a highly pure product, which was identified by polyacrylamide gel electrophoresis and amino acid analysis.     

Use of phage display methods to identify heptapeptide sequences for use as affinity purification 'tags' with novel chelating ligands in immobilized metal ion affinity chromatography

This study describes the screening of a peptide phage display library for amino acid sequences that bind with different affinities to a novel class of chelating ligands complexed with Ni2+ ions. These chelating ligands are based on the 1,4,7-triazacyclononane (TACN) structure and have been chosen to allow enhanced efficiency in protein capture and decreased propensity for metal ion leakage in the immobilized metal ion affinity chromatographic (IMAC) purification of recombinant proteins. Utilising high stringency screening conditions, various peptide sequences containing multiple histidine, tryptophan, and/or tyrosine residues were identified amongst the different phage peptide sequences isolated. The structures, and particularly the conserved locations of these key amino acid residues within the selected heptapeptides, form a basis to design specific peptide tags for use with these novel TACN ligands as a new mode of IMAC purification of recombinant proteins.     

An approach to sequence-specific antibody proteases

We describe here a novel strategy for the isolation of antibodies with sequence-specific protease activity: the synthesis of dipeptide haptens in which the targeted peptide bond has been replaced by a ring-strained or torsionally strained hydroxyethylene transition-state analog. Thus, the analogs mimic both a peptide bond in a distorted, reactive conformation and the transition state for peptide bond hydrolysis. In order to obtain sequence-specific antibody proteases, these analogs have been flanked with additional amino acid residues in preparation for immunization. In particular, we have synthesized peptides containing analogs such as 2-cis-amino-3-cis-hydroxycyclobutane carboxylic acid andendo-(3-amino-2-hydroxy)bicyclo[2.2.1]heptane-7-anti-carboxylic acid. We have also prepared a series of peptide derivatives containing analogs, such as 2-[3-amino-2-oxo-1-azetidinyl]-3-methylbutanoic acid, in which the targeted peptide bond has been incorporated into a β-lactam ring. Since the “peptide bond” has been left intact, these species mimic only a distorted ground state. At present, antibodies are being elicited against a number of the above peptide derivatives.     

Combinatorial Peptide Library for Probing the Selectivity of the s-1 Subsite of Proteases

A combinatorial tetrapeptide library, Suc-Ala-Phe-Arg-AA₁-OR, in which R = p-formamidobenzyl ester and AA₁ = 17 of the 20 natural occurring amino acids, has been synthesized chemically and separated by a reverse phase HPLC. The library was used to study the s-1 subsite specificity of various proteases. The preferred substrate at the s-1 subsite of chymotrypsin is in the order of Trp > Tyr > Phe > Met > Leu. This agreed with the reported data that the favored substrate at the s-1 subsite for chymotrypsin-catalyzed hydrolysis is an aromatic amino acid residue. The hydrophobic amino acid residues at this subsite can be hydrolysized after a longer incubating time. This procedure of selective hydrolysis of a peptide library was used to probe the selectivity of s-1 subsites of four proteases isolated from Bacillus stearothermophilus, subtilisin Carlsberg, subtilisin BPN' and an engineered protease subtilisin 8397. The protease from Bacillus stearothermophilus favored the substrate with residue Lys, and Arg at the s-1 subsite as a trypsin-like protease. The relative reactivities of amino acid residues in the protease-catalyzed hydrolysis of the library can be used as a fingerprint to identify the protease in a protease family.