Identification of genes encoding for peptide synthetases in the gram-negative bacterium Lysobacter sp. ATCC 53042 and the fungus Cylindrotrichum oligospermum

Abstract

Genes encoding for the multifunctional peptide synthetases lysobactin synthetase and peptolide SDZ 214-103 synthetase were identified by hybridization of genomic libraries with oligonucleotides derived from consensus motifs of various genes encoding for β-(L-α amino-adipoyl)-L-cysteinyl-D-valine (ACV) synthetases and gramicidin S synthetase. The sequence of subcloned gene fragments revealed core motifs and a modular structure typical for the family of peptide synthetase genes. A fragment of 4.6 kb of the lysobactin synthetase gene was sequenced and one amino acid activating module was localized. The cloning of lysobactin synthetase was verified by marker-exchange mutagenesis and the lysobactin minus phenotype of the mutant. The sequenced 3.1 kb fragment of peptolide SDZ 214-103 synthetase contained parts of two modules and was highly homologous to corresponding regions of module 6 and 7 of cyclosporin synthetase. Therefore, the localized modules may activate the amino acids threonine and glycine.

Key Words:

• Peptide. Lactone Antibiotics
• Gliding Bacteria
• Filamentous Fungi