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Journal of Bacteriology, November 2000, p. 6233-6238, Vol. 182, No. 21
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Pseudobactin Biogenesis in the Plant Growth-Promoting Rhizobacterium Pseudomonas Strain B10: Identification and Functional Analysis of the L-Ornithine N5-Oxygenase (psbA) Gene

C. Ambrosi,1 L. Leoni,2 L. Putignani,1 N. Orsi,1 and P. Visca2,3,*

Istituto di Microbiologia, Università di Roma "La Sapienza",1 Dipartimento di Biologia, Università di Roma Tre,2 and Unità di Microbiologia Molecolare, I.R.C.C.S. "Lazzaro Spallanzani",3 00100 Rome, Italy

Received 27 March 2000/Accepted 1 August 2000

PseudobactinB10, the fluorescent siderophore produced by the rhizobacterium Pseudomonas strain B10, contains the hydroxamate ligand D-N5-hydroxyornithine (D-N5-OH-Orn). We cloned the L-Orn N5-oxygenase (psbA) gene from a genomic library of Pseudomonas strain B10 and demonstrated that PsbA is involved in the conversion of L-Orn to its N5-OH derivative. PsbA shows significant similarity to microbial omega -amino acid hydroxylases containing flavin adenine dinucleotide and NADP cofactor-binding sites and the FATGY signature of the putative substrate recognition pocket. The psbA gene is monocistronic, and its transcription is negatively controlled by iron. A site-specific psbA mutant of Pseudomonas strain B10 was biochemically complemented with the precursor L-N5-OH-Orn, suggesting that L-Orn is hydroxylated before conversion to the D isomer. The L-Orn N5-hydroxylase-defective mutants of Pseudomonas strain B10 and Pseudomonas aeruginosa PAO1 were much less effective than the parental strains in suppressing the growth of the phytopathogen Erwinia carotovora in iron-poor medium. The extent of in vitro inhibition of E. carotovora was strictly iron dependent and directly correlated with the amount of released siderophores. These data strengthen the role of fluorescent siderophores in biocontrol of deleterious rhizomicroorganisms.

* Corresponding author. Mailing address: Dipartimento di Biologia, Università di Roma Tre, Viale G. Marconi 446, 00146 Rome, Italy. Phone: 39.6.5517.6331. Fax: 39.6.5517.6321. E-mail: visca{at}bio.uniroma3.it.

Journal of Bacteriology, November 2000, p. 6233-6238, Vol. 182, No. 21
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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