title: A hybrid non-ribosomal peptide/polyketide synthetase containing fatty-acyl ligase (FAAL) synthesizes the β-amino fatty acid lipopeptides puwainaphycins in the Cyanobacterium Cylindrospermum alatosporum.
abstract: A putative operon encoding the biosynthetic pathway for the cytotoxic cyanobacterial lipopeptides puwainphycins was identified in Cylindrospermum alatosporum. Bioinformatics analysis enabled sequential prediction of puwainaphycin biosynthesis; this process is initiated by the activation of a fatty acid residue via fatty acyl-AMP ligase and continued by a multidomain non-ribosomal peptide synthetase/polyketide synthetase. High-resolution mass spectrometry and nuclear magnetic resonance spectroscopy measurements proved the production of puwainaphycin F/G congeners differing in FA chain length formed by either 3-amino-2-hydroxy-4-methyl dodecanoic acid (4-methyl-Ahdoa) or 3-amino-2-hydroxy-4-methyl tetradecanoic acid (4-methyl-Ahtea). Because only one puwainaphycin operon was recovered in the genome, we suggest that the fatty acyl-AMP ligase and one of the amino acid adenylation domains (Asn/Gln) show extended substrate specificity. Our results provide the first insight into the biosynthesis of frequently occurring β-amino fatty acid lipopeptides in cyanobacteria, which may facilitate analytical assessment and development of monitoring tools for cytotoxic cyanobacterial lipopeptides.
authors.affiliation: Institute of Microbiology AS CR, v.v.i., Department of Phototrophic Microorganisms - ALGATECH, Třeboň, Czech Republic; Biology Centre of AS CR, v.v.i., Institute of Hydrobiology, České Budějovice, Czech Republic; University of South Bohemia, Faculty of Science, Department of Botany, České Budějovice, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Mareš
authors.affiliation: Institute of Microbiology AS CR, v.v.i., Department of Phototrophic Microorganisms - ALGATECH, Třeboň, Czech Republic; Biology Centre of AS CR, v.v.i., Institute of Hydrobiology, České Budějovice, Czech Republic; University of South Bohemia, Faculty of Science, Department of Molecular Biology and Genetics, České Budějovice, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Hájek
authors.affiliation: Institute of Microbiology AS CR, v.v.i., Department of Phototrophic Microorganisms - ALGATECH, Třeboň, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Urajová
authors.affiliation: Institute of Microbiology AS CR, v.v.i., Department of Phototrophic Microorganisms - ALGATECH, Třeboň, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Kopecký
authors.affiliation: Institute of Microbiology AS CR, v.v.i., Department of Phototrophic Microorganisms - ALGATECH, Třeboň, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Hrouzek
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology; D053898:Biosynthetic Pathways; D000458:Cyanobacteria / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D008025:Ligases / Q000235:genetics* / Q000502:physiology; D055666:Lipopeptides / Q000096:biosynthesis; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D005810:Multigene Family; D048630:Polyketide Synthases / Q000235:genetics* / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D055666:Lipopeptides; C577099:puwainaphycin F; C577100:puwainaphycin G; D048630:Polyketide Synthases; D008025:Ligases