Whole cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and in situ structure analysis of streptocidins, a family of tyrocidine-like cyclic peptides.

Streptocidins, a family of tyrocidine-like cyclic decapeptides, are an ideal demonstration object for the detection and in situ structure analysis of natural compounds directly in microbial cells using whole cell matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-TOFMS), an emerging technique that can be used for rapid sensitive metabolic profiling of microorganisms. Five main members of the streptocidin family (A-E) were detected in Brevibacillus cells picked from agar plates and identified by in situ structure analysis with post-source decay MALDI-TOFMS. This efficient modern method allows the precise detection of metabolites within minutes without the need to isolate and purify the target compounds. The generated mass spectra are of similar quality to those obtained for the purified peptides. In addition, surface extracts were prepared by treating Brevibacillus cells with 70% acetonitrile in the presence of 0.1% trifluoroacetic acid and fractionated by high-resolution reversed-phase high-performance liquid chromatography (HPLC). In this way ten minor streptocidins were detected demonstrating the full biosynthetic variety of streptocidin production on the cellular level. The streptocidins differ from the well-known tyrocidines essentially in position 3 of the decapeptide chain by replacement of the aromatic amino acid (F/W) found in tyrocidines by L-leucine or L-valine.

[1]  T. Stein,et al.  Rapid typing of Bacillus subtilis strains by their secondary metabolites using matrix‐assisted laser desorption/ionization mass spectrometry of intact cells , 1999 .

[2]  L. Craig,et al.  THE CHEMISTRY OF TYROCIDINE. VI. THE AMINO ACID SEQUENCE OF TYROCIDINE C. , 1965, Biochemistry.

[3]  Christopher Wilde,et al.  "Whole cell"--matrix-assisted laser desorption ionization-time of flight-mass spectrometry, an emerging technique for efficient screening of biocombinatorial libraries of natural compounds-present state of research. , 2003, Combinatorial chemistry & high throughput screening.

[4]  E. Katz,et al.  The peptide antibiotics of Bacillus: chemistry, biogenesis, and possible functions. , 1977, Bacteriological reviews.

[5]  L. Craig,et al.  The Chemistry of Tyrocidine. III. The Structure of Tyrocidine A , 1954 .

[6]  J. Hacker,et al.  Antimicrobial Activities and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry of Bacillus Isolates from the Marine Sponge Aplysina aerophoba , 2003, Marine Biotechnology.

[7]  B. Mach,et al.  Studies on amino acid substitution in the biosynthesis of the antibiotic polypeptide tyrocidine. , 1966, Biochemistry.

[8]  J. Lay,et al.  MALDI-TOF mass spectrometry of bacteria. , 2001, Mass spectrometry reviews.

[9]  H. von Döhren,et al.  Nonribosomal biosynthesis of peptide antibiotics. , 1990, European journal of biochemistry.

[10]  M. Erhard,et al.  Rapid typing and elucidation of new secondary metabolites of intact cyanobacteria using MALDI-TOF mass spectrometry , 1997, Nature Biotechnology.

[11]  T. Stein,et al.  4.13 – Structure, Function, and Biosynthesis of Gramicidin S Synthetase , 1999 .

[12]  S. Cameotra,et al.  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 , 2002, Applied and Environmental Microbiology.

[13]  M. Erhard,et al.  Rapid identification of the new anabaenopeptin G from Planktothrix agardhii HUB 011 using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. , 1999, Rapid communications in mass spectrometry : RCM.

[14]  G. Jung,et al.  Streptocidins A-D, novel cyclic decapeptide antibiotics produced by Streptomyces sp. Tü 6071. II. Structure elucidation. , 2001, The Journal of antibiotics.

[15]  M. Landy,et al.  Bacillomycin; an antibiotic from Bacillus subtilis active against pathogenic fungi. , 1948, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[16]  R. Roskoski,et al.  Purification of the polyenzymes responsible for tyrocidine synthesis and their dissociation into subunits. , 1973, Biochemistry.

[17]  L. Craig,et al.  The Chemistry of Tyrocidine. V. The Amino Acid Sequence of Tyrocidine B , 1955 .

[18]  T. Suzuki,et al.  Biosynthesis of tyrocidine by a cell-free enzyme system of Bacillus brevis ATCC 8185. II. Amino acid substitution in tyrocidine. , 1968, Biochimica et biophysica acta.

[19]  R. Pukall,et al.  Streptocidins A-D, novel cyclic decapeptide antibiotics produced by Streptomyces sp. Tü 6071. I. Taxonomy, fermentation, isolation and biological activities. , 2001, The Journal of antibiotics.

[20]  R. Roskoski,et al.  Tyrocidine biosynthesis by three complementary fractions from Bacillus brevis (ATCC 8185). , 1970, Biochemistry.

[21]  B. V. Baar.  Characterisation of bacteria by matrix-assisted laser desorption/ionisation and electrospray mass spectrometry. , 2000 .

[22]  Zhihong Guo,et al.  Facile solid-phase synthesis of cyclic decapeptide antibiotic streptocidins A–D , 2004 .

[23]  M. Erhard,et al.  Determination of Oligopeptide Diversity within a Natural Population of Microcystis spp. (Cyanobacteria) by Typing Single Colonies by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry , 2001, Applied and Environmental Microbiology.

[24]  R. Roskoski,et al.  Polypeptide biosynthesis form thioesters of amino acids. , 1971, Archives of biochemistry and biophysics.

[25]  R. Andersen,et al.  Loloatins A-D, cyclic decapeptide antibiotics produced in culture by a tropical marine bacterium. , 1999, Journal of natural products.