Unit of Microbiology and Biotechnology, Vector Control Research Centre, Indian Council of Medical Research, Indira Nagar, Puducherry, 605 006, India.
Appl Microbiol Biotechnol. 2010 May;86(6):1737-44. doi: 10.1007/s00253-010-2449-y. Epub 2010 Feb 4.
The culture supernatant of a strain of Bacillus subtilis subsp. subtilis isolated from mangrove forests of Andaman and Nicobar islands, India was found to kill larval and pupal stages of mosquitoes. A chloroform extract of the culture supernatant of the bacterium showed pupicidal effects at an LC(50) dose of 1 microg/ml. The mosquitocidal metabolite(s) produced by this strain were purified by gel permeation chromatography. The purified fraction was subjected to Fourier transform infrared (FTIR) spectroscopy and Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. The FTIR spectrum of active fraction/CHCl3 residue showed strong band characteristic of peptides. MALDI-TOF spectrum of the sample showed well-resolved group of peaks at m/z values 1,030.6, 1,046.7, 1,044.6, 1,060.5, 1,058.6, 1,058.7, and 1,074.6. The results indicated production of different isoforms of surfactin, ranging from C13-C15. Further, the sfp gene responsible for the production of surfactin was amplified and sequenced. In conclusion, this study showed that the mosquito pupicidal metabolite(s), produced by B. subtilis subsp. subtilis is the cyclic lipopeptide, surfactin. The mode of action of surfactin on pupae of mosquitoes is discussed. This is the first report on the mosquito pupicidal activity of surfactin produced by B. subtilis subsp. subtilis.
从印度安达曼和尼科巴群岛的红树林中分离到的一株枯草芽孢杆菌亚种的培养物上清液被发现能杀死蚊子的幼虫和蛹期。该细菌培养物上清液的氯仿提取物在 LC(50)剂量为 1μg/ml 时显示出杀蛹作用。该菌株产生的杀蚊代谢物通过凝胶渗透色谱进行了纯化。纯化的部分进行了傅里叶变换红外（FTIR）光谱和基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱分析。活性部分/CHCl3 残留物的 FTIR 光谱显示出强烈的肽特征带。样品的 MALDI-TOF 光谱显示出在 m/z 值 1,030.6、1,046.7、1,044.6、1,060.5、1,058.6、1,058.7 和 1,074.6 处有很好分辨的峰群。结果表明，产生了不同的表面活性肽异构体，范围从 C13-C15。此外，还扩增和测序了 sfp 基因，该基因负责表面活性肽的产生。总之，本研究表明，枯草芽孢杆菌亚种产生的蚊子杀蛹代谢物是环状脂肽表面活性肽。讨论了表面活性肽对蚊子蛹的作用方式。这是枯草芽孢杆菌亚种产生的表面活性肽对蚊子杀蛹活性的首次报道。
