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Genetic and functional characterization of the gene cluster directing the biosynthesis of putisolvin I and II in Pseudomonas putida strain PCL1445

¹ Institute of Biology, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands
² Institute of Infection, Immunity & Inflammation, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG27 2RD, UK
³ Wageningen University, Laboratory of Bioinformatics, Genome Informatics Group, PO Box 8128, 6700 ET Wageningen, The Netherlands
⁴ Institute of Medical Microbiology, University of Zürich, Gloriastr. 32, CH-8006 Zürich, Switzerland

Correspondence
Guido V. Bloemberg
bloemberg{at}immv.uzh.ch

Pseudomonas putida PCL1445 secretes two cyclic lipopeptides, putisolvin I and putisolvin II, which possess a surface-tension-reducing ability, and are able to inhibit biofilm formation and to break down biofilms of Pseudomonas species including Pseudomonas aeruginosa. The putisolvin synthetase gene cluster (pso) and its surrounding region were isolated, sequenced and characterized. Three genes, termed psoA, psoB and psoC, were identified and shown to be involved in putisolvin biosynthesis. The gene products encode the 12 modules responsible for the binding of the 12 amino acids of the putisolvin peptide moiety. Sequence data indicate that the adenylation domain of the 11th module prioritizes the recognition of Val instead of Leu or Ile and consequently favours putisolvin I production over putisolvin II. Detailed analysis of the thiolation domains suggests that the first nine modules recognize the D form of the amino acid residues while the two following modules recognize the L form and the last module the L or D form, indifferently. The psoR gene, which is located upstream of psoA, shows high similarity to luxR-type regulatory genes and is required for the expression of the pso cluster. In addition, two genes, macA and macB, located downstream of psoC were identified and shown to be involved in putisolvin production or export.

The GenBank/EMBL/DDBJ accession number for the nucleotide sequence of the P. putida PCL1445 oprM-psoR-psoA-psoB-psoC-macA-macB DNA region is DQ151887.