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Identification and characterization of a mosquito pupicidal metabolite of a Bacillus subtilis subsp. subtilis strain

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Abstract

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 LC50 dose of 1 µg/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.

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Acknowledgments

We are indebted to Dr. P. Jambulingam, Director, Vector Control Research Centre, Dr. K. Balaraman (Retd)., Dr. S. L. Hoti, Scientist “F,” for their suggestions, encouragement, and the facilities provided throughout the study. The technical assistance rendered by Mr. S. Venugopalan is gratefully acknowledged. We thank Dr. Nisha Mathew, Scientist “C,” for FTIR spectroscopy. We would also like to thank Dr. Joachim Vater, Institut für Chemie, Arbeitsgruppe Biochemie und Molekulare Biologie, Technische Universität Berlin for MALDI-TOF analysis of the sample. This study was supported by a research grant from the Department of Science and Technology (DST), Government of India.

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  1. Unit of Microbiology and Biotechnology, Vector Control Research Centre, Indian Council of Medical Research (ICMR), Indira Nagar, Puducherry, 605 006, India

    I. Geetha, A. M. Manonmani & K. P. Paily

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  1. I. Geetha
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  2. A. M. Manonmani
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  3. K. P. Paily
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Correspondence to I. Geetha.

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Geetha, I., Manonmani, A.M. & Paily, K.P. Identification and characterization of a mosquito pupicidal metabolite of a Bacillus subtilis subsp. subtilis strain. Appl Microbiol Biotechnol 86, 1737–1744 (2010). https://doi.org/10.1007/s00253-010-2449-y

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