Biosynthesis of isotopically labeled gramicidins and tyrocidins by Bacillus brevis |
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Authors: | Vogt T C Bas Schinzel Susan Bechinger Burkhard |
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Institution: | (1) Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, 82152 Martinsried, Germany;(2) Université Strasbourg, Faculté de chimie, Institut Le Bel, 4 rue Blaise Pascal, F-67000 Strasbourg, France |
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Abstract: | The three-dimensional structure of bilayer-associated gramicidin A is available from a structural data base. This and related peptides are, therefore, ideal model compounds to use during the implementation and development of new NMR techniques for the structural investigations of membrane proteins. As these methods rely on the isotopic labelling of single, selected or all sites, we have, investigated and optimised biochemical protocols using different strains of the Gram-positive bacterium Bacillus brevis. With newly developed schemes for isotopic labelling large amounts of gramicidin and tyrocidin enriched with stable isotopes such as 15N or 15N/13C have been obtained at low cost. A variety of analytical and spectroscopic techniques, including HPLC, mass spectrometry and NMR spectroscopy are used to characterise the resulting products. |
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Keywords: | antibiotic peptide fermentation and purification of peptides high-resolution NMR oriented lipid bilayer solid-state NMR structure determination |
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