混合氨基酸月桂酰胺的合成

本文以玉米淀粉生产副产物为原料,提取蛋白质水解制取混合氨基酸,与酰化剂月桂酰氯合成混合氨基酸月桂酰胺表面活性剂,研究了反应条件,并测定了它的pH值、表面张力、乳化力等。     

一种构建高质量随机肽库的有效方法

为构建完全随机化的基因工程肽库 ,克服现有简并方案中终止密码子和序列组成偏歧的不足 ,提出了一种新的简并DNA文库合成方式。通过这种分组式合成方式构建的肽库可以避免终止密码子的出现和氨基酸组成偏歧的发生 ,还可以控制随机化过程中不同氨基酸的参入比例。以一个 13肽库的合成过程为例对分组式合成法进行了实验 ,测序结果和对 19种氨基酸出现频率的统计表明没有终止密码子和半胱氨酸密码子出现 ,各氨基酸的出现频率接近均值 ,表明这种分组 混合 分组 ,辅以简并合成的方法是行之有效的 ,能满足各类高容量基因工程随机肽库要求。     

生长激素释放肽的合成及促生长活性

用固相多肽合成法合成了生长激素释放肽(GHRP),这是一种含D-型氨基酸的外源性激素,具有促进脑下垂体分泌生长激素功能的人工合成六肽,其氨基酸序列为His-D-Trp-Ala-Trp-D-Phe-Lys-NH₂.观察了使用不同剂量的GHRP对不同日龄小鼠的促生长效应,当使用最佳剂量(1000μg/kg)时,可使25日龄小鼠体重较对照组增加14.8%,同时发现鼠龄越小对GHRP越敏感,当使用剂量高达10mg/kg时仍然安全无毒.     

15种氨基酸随机肽库的小盒式DNA编码文库的构建

采用小盒式DNA编码文库的构建策略,选取在进化上可能起源较早的15种氨基酸,按照其简并密码子合成了一个为10个随机氨基酸编码的小盒式DNA模板,经过连续3轮的PCR扩增、酶切及连接的小盒式文库组装过程,成功构建了一个文库容量达1.31×10¹²/ml,随机编码区长达97个氨基酸的小盒式DNA编码文库。     

降钙素基因相关肽的合成

降钙素基因相关肽是一个包含有37个氨基酸残基的具有较强降血压生理功能的活性多肽。我们采用常规的固相合成方法,以简单的装置最后经无水氟化氢处理,将肽从树脂上切下,同时脱除所有侧链保护基。粗产物在30％乙酸溶液中用碘作为氧化剂使二个半胱氨酸氧化,形成二硫键。合成的α-人降钙素基因相关肽经反向高效液相层析分离,获得在高效液相层析为单一峰的产物。经酸水解氨基酸分析证明与理论值相符并具有全部生理活性。     

利用噬菌体随机肽库筛选抗柯萨奇病毒B3多肽的研究

本实验将柯萨奇病毒B3型(CVB3)大量扩增,应用蔗糖密度梯度离心法纯化病毒.利用噬菌体随机9肽库进行筛选,3轮淘洗后,测定噬菌体克隆抗病毒复制能力.提取阳性克隆DNA并进行测序,推导外源多肽的氨基酸序列.结果表明3个具有明显抗病毒复制能力的噬菌体阳性克隆被筛选出来,使TCID50由10-7.5SFU/mL分别降至10-5.25、10-6、10-5.5SFU/mL,由此证明可以应用噬菌体肽库来筛选具有抗病毒作用的多肽,本研究为抗病毒多肽制剂的研究奠定了基础.     

利用噬菌体随机肽库筛选抗柯萨奇病毒B3多肽的研究

本实验将柯萨奇病毒B3型(CVB3)大量扩增,应用蔗糖密度梯度离心法纯化病毒。利用噬菌体随机9肽库进行筛选,3轮淘洗后,测定噬菌体克隆抗病毒复制能力。提取阳性克隆DNA并进行测序,推导外源多肽的氨基酸序列。结果表明:3个具有明显抗病毒复制能力的噬菌体阳性克隆被筛选出来,使TCID50由10-7.5SFU/mL分别降至10-5.25、10-6、10-5.5SFU/mL,由此证明可以应用噬菌体肽库来筛选具有抗病毒作用的多肽,本研究为抗病毒多肽制剂的研究奠定了基础。     

人胰岛素原C肽的合成及专一性抗体制备

利用固相多肽合成中的Boc途径合成了人胰岛素原C肽 ,经TSK柱层析一步纯化 ,产物达HPLC及毛细管电泳均一 ,蛋白质序列分析和质谱分析符合理论值 ,总回收率高达 41 %。为了提高寡肽抗体的专一性 ,我们将丙烯酰化的C肽经催化聚合形成了以聚丙酰为骨架 ,C肽为侧链的聚合体 ,分子量约为 2 5kD。免疫新西兰大白兔 1月 ,获得专一性抗体。用酶联免疫吸附法测得抗血清滴度达 2 .5× 1 0 4 ,与BSA无交叉反应。聚丙酰C肽抗原是制备C肽抗体的一种新的尝试 ,而且也可能成为新的多肽工程疫苗之一 ,为其他传染性疾病多肽疫苗的研究提供新的途径     

The Synthesis of Phosphopeptides Using Microwave-assisted Solid Phase Peptide Synthesis

A series of phosphorylated peptides were synthesised using microwave mediated solid phase peptide synthesis. Acidic cleavage of peptides from the solid support using microwave irradiation often resulted in reattachment of the phosphate benzyl protecting group to the peptide chain. However for most phosphopeptide sequences performing the cleavage reaction at room temperature in order to minimize this undesired alkylation was successful. Notably for phosphopeptides containing a methionine residue flanking the phosphorylated residue (for serine and threonine) the trifluoroacetic acid mediated cleavage afforded the benzylated side product as a major component. This detrimental process was not observed for a corresponding tyrosine containing sequence.     

Synthesis and Screening of an Indexed Motif-Library Containing Non-proteinogenic Amino Acids

In an effort to increase the probability of finding novel peptides in resin-bound combinatorial libraries displaying affinity to various macromolecular targets, we increased the diversity of a solid-phase library considerably by synthesizing multiple structures on each bead – a motif-library – including 45 building blocks. The building blocks consist of L -aa, D -aa and eight hydrophobic non-proteinogenic α-amino acids. A library with the format O-Z_0–1-O-Z_0–1-O-XX-resin was synthesized giving the four motifs OOOXX, OZOOXX, OOZOXX, OZOZOXX corresponding to 364.500 different motifs (45³×4 theoretical combinations). The positions O are defined amino acids while Z represents three mixtures Π, Ω, ϖ, where Π is a mixture of polar and charged residues, Ω is a mixture of aliphatic residues and ϖ is a mixture of aromatic residues. X represents a mixture of all 45 residues. The library was screened with the macromolecular target streptavidin which served as a model receptor. Binding peptides were sequenced by microsequencing. We included small amounts of norvaline and norleucine in the library, which served as index residues to be able to distinguish between LD -amino acids and other residues with the same retention time in the HPLC system. Beads that interact with the receptor were found, and the binding motifs that appeared had no homology to known binding motifs found in either L -aa or D -aa libraries, instead motifs with the non-proteinogenic residues L -phenylglycine, O-benzyl-L -hydroxyproline and O-benzyl-L -tyrosine dominated. The novel peptides inhibit binding of biotin to streptavidin but do not bind to avidin, and the affinity is higher than the peptides found in linear all L -aa peptide libraries. © 1997 European Peptide Society and John Wiley & Sons, Ltd     

Synthesis of Fragments of the Peptide Component of Pseudobactin

Pseudobactin is a structurally complex and physiologically important siderophore (microbial iron chelator) from Pseudomonas putida- fluorescens. Various fragments of the unusual peptide component of pseudobactin listed below were prepared by solution-phase peptide synthesis. L -Lys· D -threo-β-OH Asp· L -Ala· D -allo-Thr· L -Ala L -Lys· D -threo-βOH Asp· L -Ala· D -allo-Thr D -threo-β-OH Asp· L -Ala· D - allo-Thr· L -Ala· D -N-OH-cycloOrn D -threo-β-OH-Asp· L -Ala· D -allo-Thr· L -Ala L -Ala· D -allo-Thr· L - Ala· D -N-OH-cycloOrn A class of related peptides named pseudomycins have shown promising antifungal activity. To examine if these peptide fragments above would elicit similar activity, the fragments were tested and found to have no antifungal activity in limited bioassays.     

支链氨基酸对运动大鼠氨基酸代谢和运动能力的影响

观察了支链氨基酸(BCAA)对大鼠运动能力和血清游离氨基酸代谢的影响。实验用21只雄性wistar大鼠,随机分为3组:正常组、游泳对照组和游泳补充BCAA组。2个运动组每天游泳训练1h,10天后游泳6h,观察补充BCAA对大鼠游泳运动能力和血清游离氨基酸水平的影响。实验结果表明,补充BCAA可明显提高大鼠游泳存活率,抑制血清中必需氨基酸、非必需氨基酸和总氨基酸水平升高,游泳运动后大鼠的血清中乳酸和LDH的升高幅度有所降低,抑制骨骼肌LDH活力的下降。说明补充BCAA可明显提高大鼠的运动能力,减少运动造成的蛋白质分解     

氨基酸对心血管功能的影响

介绍了氨基酸对心血管功能的保护作用,氨基酸不仅可减轻心肌在缺血再灌注、心力衰竭和心肌梗塞中的损伤,而且对血压、血脂和心律失常也有一定的调节作用。     

提取法生产氨基酸的最新技术进展

对提取法生产氨基酸的工艺路线、技术状况、工艺设备及我国采用提取法从毛发中生产氨基酸行业的最新技术进展进行了总结和展望。     

Bradykinin in Solid-phase Peptide Synthesis

Bradykinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg; BK) occupies a special place in the history of the development of solid-phase peptide synthesis (SPPS). It was the first biologically-active peptide to be synthesized by Merrifield using the new method. Since that time many hundreds of BK analogs have been synthesized by SPPS. Certain of these analogs show potential as drug candidates. Dedicated to the memory of Bruce Merrifield. John Stewart (left) and RB Merrifield (right) with the first automatic peptide synthesizer.      

从鸡毛中提取复合氨基酸的研究

运用正交实验研究了以鸡毛为原料硫酸水解生产复合氨基酸的水解条件。结果表明 :硫酸浓度为 8mol·L- 1 ,鸡毛质量 (g)与硫酸体积 (mL)之比为 1∶2 .5 ,水解时间 10h ,其水解率可达 5 5 .2 3%。得到的固体氨基酸质量分数为 95 .31%。     

乙内酰脲酶及其在氨基酸手性合成中的应用

乙内酰脲水解酶、氨甲酰化酶和乙内酰脲消旋酶构成的酶系能够以5-取代乙内酰脲类化合物为原料合成天然和非天然D-或L-氨基酸,用于各种手性氧基酸的生产。近来的研究重点在分离新酶或提高原酶的活性,包括定向突变、三维结构解析与结构功能关系研究、酶固定化、蛋白融合和构建完整细胞生物催化剂等。