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Applied and Environmental Microbiology, December 2002, p. 6210-6219, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6210-6219.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry of Lipopeptide Biosurfactants in Whole Cells and Culture Filtrates of Bacillus subtilis C-1 Isolated from Petroleum Sludge

Joachim Vater,^1* Bärbel Kablitz,¹ Christopher Wilde,¹ Peter Franke,² Neena Mehta,³ and Swaranjit Singh Cameotra³

Institut für Chemie, Arbeitsgruppe Biochemie und Molekulare Biologie, Technische Universität Berlin, D-10587 Berlin,¹ Institut für Biochemie, Freie Universität Berlin, D-14195 Berlin, Germany,² Institute of Microbial Technology, Sector 39A, Chandigarh-160 036, India³

Received 6 May 2002/ Accepted 1 August 2002

An innovative method was developed for rapid sensitive detection and efficient structural characterization of lipopeptide biosurfactants by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry by using whole microbial cells and crude culture filtrates as targets in combination with surface tension measurements. This was done for a bacterial strain that was isolated from petroleum sludge and efficiently produces biosurfactants. This organism was identified by using biochemical, physiological, and genetic parameters as a Bacillus subtilis strain, designated B. subtilis C-1. This assignment was supported by a mass spectrometric investigation of the secondary metabolite spectrum determined by whole-cell MALDI-TOF mass spectrometry, which revealed three lipopeptide complexes, the surfactins, the iturins, and the fengycins, which are well-known biosurfactants produced by B. subtilis strains. These compounds were structurally characterized by in situ structure analysis by using postsource decay MALDI-TOF mass spectrometry. The isoforms were separated by miniaturized high-resolution reversed-phase high-performance liquid chromatography for mass spectrometric characterization. Iturin compounds which contain unusual fatty acid components were detected.

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* Corresponding author. Mailing address: Institut für Chemie, Arbeitsgruppe Biochemie und Molekulare Biologie, Technische Universität Berlin, Franklinstr. 29, D-10587 Berlin, Germany. Phone: 49-30-31425609. Fax: 49-30-31424783. E-mail: Vater{at}chem.tu-berlin.de.

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Applied and Environmental Microbiology, December 2002, p. 6210-6219, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6210-6219.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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