蜂毒肽衍生物的固相合成与表征
Solid-phase synthesis and characterization of melittin derivatives
张翔 1黄磊 1宫玉坤 1陈维晟 1柯莹莹 1汪燕 1王爱东1
作者信息
- 1. 黄山学院化学化工学院，黄山 245041
- 折叠
摘要
蜂毒肽（Melittin）是蜜蜂毒素的主要活性衍生多肽，由26个氨基酸残基组成，具有抗炎、抗菌等多种生物活性。本研究采用采用Fmoc固相合成策略（SPPS），用全自动多肽合成仪合成蜂毒肽，以 Rink Amide MBHA 树脂为载体、HBTU/DIEA 为缩合体系，由C端至N端逐次偶联氨基酸合成蜂毒肽，通过Kaiser试验与四氯苯醌试验监测脱保护及缩合反应进程。对 Fmoc 脱除条件进行系统优化，确定适宜的反应温度与时间；采用 TFA 混合裂解液将多肽从树脂上切割，经乙醚沉淀、洗涤、冷冻干燥获得粗肽。建立蜂毒肽高效液相色谱标准曲线，实现对目标产物的准确定量。产物经高效液相色谱（HPLC）、傅里叶变换红外光谱（FT-IR）等表征，纯度良好、结构与蜂毒肽一致，同时ABTS+抗氧化实验表明，制备的蜂毒肽具有明显自由基清除能力，活性随浓度升高而增强。本研究建立了稳定可行的蜂毒肽固相合成与检测方法，为其结构修饰与活性研究提供实验依据。
Abstract
Melittin is the main biologically active polypeptide derived from bee venom, composed of 26 amino acid residues.It possesses multiple biological activities including anti-inflammatory and antibacterial effects. In this study, the Fmoc solid-phase peptide synthesis (SPPS) strategy was adopted, and melittin was synthesized by an automatic peptide synthesizer. With Rink Amide MBHA resin as the carrier and HBTU/DIEA as the condensation system, amino acids were coupled sequentially from the C-terminus to theN-terminus to synthesize melittin. The Kaiser test and chloranil test were used to monitor the progress of deprotection and condensation reactions. The Fmoc deprotection conditions were systematically optimized to determine the optimal reaction temperature and duration. A mixed trifluoroacetic acid (TFA) cleavage solution was used to cleave the polypeptide from the resin, and crude melittin was obtained via diethyl ether precipitation, washing and freeze-drying. A high-performance liquid chromatography (HPLC) standard curve of melittin was established to achieve accurate quantification of the target product. The product was characterized by HPLC and Fourier transform infrared spectroscopy (FT-IR), which verified its high purity and structural consistency with natural melittin. Meanwhile,ABTS? antioxidant assay results demonstrated that the prepared melittin exhibited significant free radical scavenging activity in a concentration-dependent manner. This study established a stable and feasible method for the solid-phase synthesis and detection of melittin, providing an experimental basis for its structural modification and further bioactivity research.关键词
蜂毒肽/固相合成/Kaiser试验/Fmoc策略/结构表征Key words
Melittin/solid-phasepeptide synthesis/Kaiser test/Fmocstrategy/structural characterization引用本文复制引用
张翔,黄磊,宫玉坤,陈维晟,柯莹莹,汪燕,王爱东.蜂毒肽衍生物的固相合成与表征[EB/OL].(2026-05-15)[2026-05-18].http://www.paper.edu.cn/releasepaper/content/202605-81.学科分类
生物科学研究方法、生物科学研究技术
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