IDENTIFICATION AND MODE OF ACTION OF ANTIBACTERIAL COMPONENTS FROM MELASTOMA MALABATHRICUM LINN LEAVES
IDENTIFICATION AND MODE OF ACTION OF ANTIBACTERIAL COMPONENTS FROM MELASTOMA MALABATHRICUM LINN LEAVES
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DOI：
10.3844/ajidsp.2013.46.58
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发表时间：
2013-01-01
期刊：
影响因子：
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通讯作者：
Ahmad, Wan
中科院分区：
文献类型：
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作者：
Alwash, Mourouge;Ibrahim, Nazlina;Ahmad, Wan
本研究的目的是评估的甲醇提取物Malabathricum叶（MMML）的抗菌活性，M。malabathricum leaves（ML 5）和分离的山奈酚-3-O-（2”，6”-di-O-p-trans-coumaroyl）-beta-glucopyranoside（Kf），并描述对革兰氏阳性和革兰氏阴性细菌的初步作用模式。MMML提取物通过真空液相色谱（VLC）分级以提供M。malabathricum叶级分（ML 1-ML 6）的极性增加。生物测定引导的分级分离（直接TLC生物自显影）显示M. malabathricum Leaves（ML 5）的抑菌活性最高。因此，将ML 5进一步进行柱层析（CC），得到山奈酚-3-O-（2”，6”-二-O-对-反式-香豆酰基）-β-吡喃葡萄糖苷（Kf）。化合物的结构用H-1 NMR和C-13 NMR谱进行了鉴定。MMML、ML 5和活性成分Kf对金黄色葡萄球菌参考菌株耐甲氧西林S.采用纸片扩散法和最低抑菌浓度（MIC）法对金黄色葡萄球菌（MRSA）、11株MRSA临床分离株、3株铜绿假单胞菌临床分离株和铜绿假单胞菌参考株进行评价。采用时间杀灭试验和场发射扫描电镜初步确定作用方式。纸片扩散法和MIC测定结果表明，MMML、ML 5和Kf对所有供试菌均表现出不同程度的抗菌活性。总体上，ML 5和Kf对S.金黄色葡萄球菌和所有测试的MRSA。MMML对所有测试细菌都表现出最强的杀灭效果，而ML 5和Kf的总体性能对S.金黄色葡萄球菌和MRSA比铜绿假单胞菌。场发射扫描电子显微镜显示不规则的细菌表面，表明MMML，ML 5和Kf破坏膜的完整性。结果表明，MMML、ML 5和Kf具有潜在的抗生素替代品。
The present study aimed to assess the antibacterial activity of the methanol extract of Melastoma Malabathricum Leaves (MMML), active fraction of M. malabathricum leaves (ML5) and isolated kaempferol-3-O-(2",6"-di-O-p-trans-coumaroyl)-beta-glucopyranoside (Kf) and to describe the preliminary mode of action against Gram-positive and Gram-negative bacteria. The MMML extract was fractionated by Vacuum Liquid Chromatography (VLC) to afford M. malabathricum leaves fractions (ML1-ML6) of increasing polarities. Bioassay-guided fractionation (direct-TLC bioautography) revealed that fraction of M. malabathricum Leaves (ML5) had the highest antibacterial activity. Therefore, the ML5 was further subjected to Column Chromatography (CC) to afford Kaempferol-3-O-(2",6"-di-O-p-trans-coumaroyl)-beta-glucopyranoside (Kf). The compound structure was elucidated using data obtained from H-1 NMR and C-13 NMR spectra. Antibacterial activity of MMML, ML5 and bioactive constituents Kf against Staphylococcus aureus reference strain, Methicillin-Resistant S. Aureus reference strain (MRSA), 11 clinical MRSA isolates, 3 clinical Pseudomonas aeruginosa isolates and P. aeruginosa reference strain was evaluated through disk diffusion method and Minimum Inhibitory Concentration (MIC). The time kill assay and field emission scanning electron microscopy was used to determine the preliminary mode of action. The data obtained from disk diffusion method and MIC showed that the MMML, ML5 and Kf showed antibacterial activity against all test bacteria with various values. Generally, the ML5 and Kf exhibited the highest antibacterial activity with the largest zones of inhibition and lowest MIC values against S. aureus and all tested MRSA. The MMML exerted the strongest killing effect against all tested bacteria while the overall performance of ML5 and Kf were significantly more effective against S. aureus and MRSA than P. aeruginosa. Field emission scanning electron microscopy revealed irregular bacterial surface indicating that the MMML, ML5 and Kf disrupted the integrity of membrane. The results obtained showed that MMML, ML5 and Kf have potential alternative to antibiotics.