Identification of surfactins and iturins produced by potent fungal antagonist, Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) tree using mass spectrometry.
Identification of surfactins and iturins produced by potent fungal antagonist, Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) tree using mass spectrometry.
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DOI：
10.1007/s13205-013-0151-3
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发表时间：
2014-06
期刊：
影响因子：
2.8
通讯作者：
Keharia, Hareshkumar
中科院分区：
文献类型：
--
作者：
Pathak, Khyati V.;Keharia, Hareshkumar
榕树内生细菌枯草芽孢杆菌K1产生一种复杂的脂肽混合物，显示出强大的抗真菌活性。这些脂肽经高效液相色谱分离纯化，并用MALDI-TOF-MS和高效液相色谱-电喷雾质谱联用进行分析。在枯草芽孢杆菌K1无细胞胞外提取液中检测到属于3个不同的环脂肽家族的脂肽的异质性混合物，即丰阳素、伊壁菌素和表面蛋白。详细的质谱学表征表明，从C13到C17的β-氨基酸链长度不同，存在四个变种的ITURIN A和三个变种的ITURIN C。一价碱金属离子加合物(Na和K)的MS/MS分析表明，Glu4是金属离子的结合部位。通过对表面活性物质的LC-ESI-MS/MS分析，可以鉴定出7种表面活性物质的变异体，它们的第4位和C13-C17位的β-羟基脂肪酸的Val/Ile/Leu发生了变化。这项研究展示了串联质谱仪在从天然来源的异质混合物中鉴定密切相关的脂肽的应用。此外，这是第一个关于内生芽孢杆菌菌株共同产生如此多的表面蛋白和iturins变体的报道。本文的在线版本(doi：10.1007/s13205-0130151-3)包含补充材料，授权用户可以使用。
The banyan endophyte, Bacillus subtilis K1, produces a complex mixture of lipopeptides exhibiting potent antifungal activity. These lipopeptides were purified by high-performance liquid chromatography and analyzed using MALDI-TOF-MS as well as liquid chromatography coupled with ESI-MS. A heterogenous mixture of lipopeptides belonging to three different families of cyclic lipopeptides, viz., fengycins, iturins and surfactins, was detected in the cell-free extracellular extract of B. subtilis K1. The detailed mass spectrometric characterization revealed the presence of four variants of iturin A and three variants of iturin C varying in the β-amino fatty acid chain length from C13 to C17. The MS/MS of monovalent alkali metal ion adducts (Na and K) of iturin suggested the Glu4 as a binding site for metal ion. The LC-ESI-MS/MS analysis of surfactins enabled the identification of seven surfactin variants with the variations in Val/Ile/Leu at position 4 and C13–C17 β-hydroxy fatty acids. This study demonstrates the application of tandem mass spectrometry in identification of closely related lipopeptides from a heterogenous mixture obtained from a natural source. Furthermore, this is the first report of an endophytic bacillus strain co-producing so many variants of surfactins and iturins. The online version of this article (doi:10.1007/s13205-013-0151-3) contains supplementary material, which is available to authorized users.
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