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Journal of Bacteriology, April 1999, p. 2166-2174, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Characterization of the Pyoluteorin Biosynthetic Gene Cluster of Pseudomonas fluorescens Pf-5

Brian Nowak-Thompson,1,2,dagger Nancy Chaney,1 Jenny S. Wing,1 Steven J. Gould,2,3,Dagger and Joyce E. Loper1,*

Agricultural Research Service, U.S. Department of Agriculture, Corvallis, Oregon 97330,1 and Department of Biochemistry and Biophysics2 and Department of Chemistry,3 Oregon State University, Corvallis, Oregon 97331

Received 27 October 1998/Accepted 19 January 1999

Ten genes (plt) required for the biosynthesis of pyoluteorin, an antifungal compound composed of a bichlorinated pyrrole linked to a resorcinol moiety, were identified within a 24-kb genomic region of Pseudomonas fluorescens Pf-5. The deduced amino acid sequences of eight plt genes were similar to the amino acid sequences of genes with known biosynthetic functions, including type I polyketide synthases (pltB, pltC), an acyl coenzyme A (acyl-CoA) dehydrogenase (pltE), an acyl-CoA synthetase (pltF), a thioesterase (pltG), and three halogenases (pltA, pltD, and pltM). Insertions of the transposon Tn5 or Tn3-nice or a kanamycin resistance gene in each of these genes abolished pyoluteorin production by Pf-5. The presumed functions of the eight plt products are consistent with biochemical transformations involved in pyoluteorin biosynthesis from proline and acetate precursors. Isotope labeling studies demonstrated that proline is the primary precursor to the dichloropyrrole moiety of pyoluteorin. The deduced amino acid sequence of the product of another plt gene, pltR, is similar to those of members of the LysR family of transcriptional activators. pltR and pltM are transcribed divergently from the pltLABCDEFG gene cluster, and a sequence with the characteristics of a LysR binding site was identified within the 486-bp intergenic region separating pltRM from pltLABCDEFG. Transcription of the pyoluteorin biosynthesis genes pltB, pltE, and pltF, assessed with transcriptional fusions to an ice nucleation reporter gene, was significantly greater in Pf-5 than in a pltR mutant of Pf-5. Therefore, PltR is proposed to be a transcriptional activator of linked pyoluteorin biosynthesis genes.

* Corresponding author. Mailing address: Horticultural Crops Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 3420 NW Orchard Ave., Corvallis, OR 97330. Phone: (541) 750-8771. Fax: (541) 750-8764. E-mail: loperj{at}bcc.orst.edu.

dagger Present address: Novartis Agribusiness Biotechnology Research, Inc., Research Triangle Park, NC 27709-2257.

Dagger Present address: Merck Research Laboratories, Rahway, NJ 07065.

Journal of Bacteriology, April 1999, p. 2166-2174, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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