A siderophore peptide synthetase gene from plant-growth-promoting Pseudomonas putida WCS358.
Under iron limiting conditions, Pseudomonas putida WCS358 produces and secretes a fluorescent siderophore called pseudobactin 358 which consists of a nonapeptide linked to a fluorescent dihydroxy quinoline moiety. Previous studies have identified a major gene cluster involved in pseudobactin 358 biosynthesis and several regulators responsible for the activation of biosynthetic genes under iron starving conditions. In this study, we identified the promoter transcribing the pseudobactin 358 synthetase gene. Promoter deletion experiments have demonstrated that the DNA region downstream of the initiation of transcription site is necessary for proper promoter functioning. This promoter controls the expression of a gene designated ppsD which encodes a 2,247-residue protein, PpsD, which has a predicted molecular weight of 247,610 Da and contains two highly homologous domains of approximately 1000 amino acids each. ppsD::Tn5 mutants of strain WCS358 are unable to synthesise pseudobactin 358 and can be complemented when ppsD is provided in trans. It is concluded that ppsD is a peptide synthetase involved in the biosynthesis of the peptide moiety of pseudobactin 358. PpsD displays a very high degree of similarity (52% aa identity) with PvdD from P. aeruginosa, a non-ribosomal peptide synthetase involved in the biosynthesis of pyoverdine, the fluorescent siderophore produced by P. aeruginosa. It also displayed homology with other peptide synthetases from other micro-organisms involved in the biosynthesis of siderophores and peptide antibiotics.
Devescovi
G
G
Bacteriology Group, International Centre for Genetic Engineering & Biotechnology, Trieste, Italy.
Aguilar
C
C
Majolini
M B
MB
Marugg
J
J
Weisbeek
P
P
Venturi
V
V
D000595:Amino Acid Sequence; D001483:Base Sequence; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D010453:Peptide Synthases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D016958:Pseudomonas putida / Q000235:genetics*; D017262:Siderophores / Q000096:biosynthesis / Q000235:genetics*
D017262:Siderophores; D010453:Peptide Synthases