Antibacterial Activity of a Cardanol from Thai Apis mellifera Propolis
Antibacterial Activity of a Cardanol from Thai Apis mellifera Propolis
复制标题
DOI：
10.7150/ijms.7373
复制
发表时间：
2014-01-01
影响因子：
3.6
通讯作者：
Chanchao, Chanpen
中科院分区：
文献类型：
--
作者：
Boonsai, Pattaraporn;Phuwapraisirisan, Preecha;Chanchao, Chanpen
背景：蜂胶是一种粘稠的，深棕色的树脂残留物，由蜜蜂从植物树脂中提取。它被用来建造和修复巢，此外还具有多种生物活性。本研究对泰国南省蜜蜂蜂胶对革兰氏（+ve）（金黄色葡萄球菌和芽孢杆菌幼虫）和革兰氏（-ve）（大肠杆菌）细菌的抗菌活性进行了测试。材料与方法：通过革兰氏染色和16S rDNA部分序列分析，证实3株分离菌属属。用甲醇、二氯甲烷和己烷依次提取蜂胶。以链霉素为阳性对照，采用琼脂孔扩散法和微汤稀释法测定其抑菌活性。通过快速柱和吸附层析进一步纯化活性最强的粗提物。采用薄层色谱法测定各生物活性部位的表观纯度。利用核磁共振（NMR）对分离得到的活性化合物进行了化学结构分析。结果：蜂胶粗甲醇提取物对金黄色葡萄球菌和大肠杆菌的最低抑菌浓度（MIC）为5 mg/mL，对P.幼虫的最低抑菌浓度（MIC）为6.25 mg/mL。快速柱层析后，只有3个活性部位对金黄色葡萄球菌和大肠杆菌的生长有抑制作用，MIC值分别为6.25和31.3 mu g/mL。进一步吸附层析得到一个纯化的生物活性组分（A1A），对大肠杆菌的IC50值为0.175 μ g/mL，对P.幼虫的IC50值为0.683 μ g/mL，经核磁共振分析确定为果仁酚。扫描和透射电镜分析显示，在腰果酚处理的大肠杆菌中，细胞形状异常（尤其是分裂细胞），细胞受损和死亡。结论：泰国蜂胶是一种很有前景的抗菌剂。
Background: Propolis is a sticky, dark brown resinous residue made by bees that is derived from plant resins. It is used to construct and repair the nest, and in addition possesses several diverse bioactivities. Here, propolis from Apis mellifera from Nan province, Thailand, was tested for antibacterial activity against Gram(+ve) (Staphylococcus aureus and Paenibacillus larvae) and Gram(-ve) (Escherichia coli) bacteria.Materials and methods: The three bacterial isolates were confirmed for species designation by Gram staining and analysis of the partial sequence of 16S rDNA. Propolis was sequentially extracted by methanol, dichloromethane and hexane. The antibacterial activity was determined by agar well diffusion and microbroth dilution assays using streptomycin as a positive control. The most active crude extract was further purified by quick column and adsorption chromatography. The apparent purity of each bioactive fraction was tested by thin layer chromatography. The chemical structure of the isolated bioactive compound was analyzed by nuclear magnetic resonance (NMR).Results: Crude methanol extract of propolis showed the best antibacterial activity with a minimum inhibition concentration (MIC) value of 5 mg/mL for S. aureus and E. coli and 6.25 mg/mL for P. larvae. After quick column chromatography, only three active fractions were inhibitory to the growth of S. aureus and E. coli with MIC values of 6.25 and 31.3 mu g/mL, respectively. Further adsorption chromatography yielded one pure bioactive fraction (A1A) with an IC50 value of 0.175 mu g/mL for E. coli and 0.683 mu g/mL for P. larvae, and was determined to be cardanol by NMR analysis. Scanning and transmission electron microscopy analysis revealed unusual shaped (especially in dividing cells), damaged and dead cells in cardanol-treated E. coli.Conclusion: Thai propolis contains a promising antibacterial agent.