Metabolic pathway monitoring of phenalinolactone biosynthesis from Streptomyces sp. Tü6071 by liquid chromatography/mass spectrometry coupling. (22nd May 2014)

• Record Type: Journal Article
• Authors: Kiske, Christiane; Erxleben, Anika; Lucas, Xavier; Willmann, Lucas; Klementz, Dennis; Günther, Stefan; Römer, Winfried; Kammerer, Bernd
• Is Part Of: Rapid communications in mass spectrometry. Volume 28:Number 13(2014)
• Page Start: 1459
• Page End: 1467
• Publication Date: 2014-05-22
• Subjects: Mass spectrometry -- Periodicals; 543.65
• DOI: 10.1002/rcm.6920
• Languages: English
• ISSNs: 0951-4198

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="rcm6920-sec-0001" sec-type="section"> <title>RATIONALE</title> <p>A rapid and precise analytical method for the investigation of natural products is required for pathway monitoring of the biosynthesis of secondary metabolites. Phenalinolactones, used in antibiotic research, are produced by <italic>Streptomyces</italic> sp. Tü6071. For the analysis of those compounds, prior to mass spectrometric analysis, an efficient separation technique is required.</p> </sec> <sec id="rcm6920-sec-0002" sec-type="section"> <title>METHODS</title> <p>For the identification of phenalinolactones from liquid cultures of <italic>Streptomyces</italic> sp. Tü6071, a new method comprising the combination of solid‐phase extraction (SPE) prior to liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI‐MS/MS) was established. MS/MS product ion scans were applied for phenalinolactone detection and structure elucidation, performed in negative mode and optimized for sensitivity and specificity. For the discovery of new intermediates, a MS/MS precursor ion scan was applied.</p> </sec> <sec id="rcm6920-sec-0003" sec-type="section"> <title>RESULTS</title> <p>Analysis of the extracts revealed that the Oasis® MAX cartridge, containing a quaternary amine functionality, is the most efficient SPE material for purification of phenalinolactones, since it allowed sufficient enrichment and detection of intermediates from the biosynthetic pathway by LC/ESI‐MS/MS. Using the precursor ion scan technique, two new secondary metabolites, PL IM1 with <italic>m/z</italic> 672.6 and PL IM2 with <italic>m/z</italic> 433.3, have been detected. The structures of the new intermediates are postulated and arranged into the biosynthetic pathway of phenalinolactones.</p> </sec> <sec id="rcm6920-sec-0004" sec-type="section"> <title>CONCLUSIONS</title> <p>A precise analytical method was established for the identification of phenalinolactones by combining purification from <italic>Streptomyces</italic> using SPE prior to LC/ESI‐MS/MS. By optimising LC/ESI‐MS/MS settings, this method has been successfully applied for pathway monitoring of secondary metabolites. Application of a precursor ion scan allowed for the identification of unknown intermediates in biosynthetic pathways. Copyright © 2014 John Wiley & Sons, Ltd.</p> </sec> </abstract>