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Title: THE PURIFICATION, CRYSTALLISATION AND PRELIMINARY STRUCTURAL CHARACTERISATION OF PHZF, A KEY ENZYME IN THE PHENAZINE BIOSYNTHESIS PATHWAY FROM PSEUDOMONAS FLUORESCENS 2-79.

Author
item MAVRODI, DMITRI - WASHINGTON STATE UNIV.
item BLEIMLING, NATHALIE - MAX-PLANK-INSTITUTE
item Thomashow, Linda
item BLANKENFELDT, WULF - MAX-PLANK-INSTITUTE

Submitted to: Acta Crystallographica Section B: Structural Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/6/2003
Publication Date: 1/20/2004
Citation: MAVRODI, D.M., BLEIMLING, N., THOMASHOW, L.S., BLANKENFELDT, W. THE PURIFICATION, CRYSTALLISATION AND PRELIMINARY STRUCTURAL CHARACTERISATION OF PHZF, A KEY ENZYME IN THE PHENAZINE BIOSYNTHESIS PATHWAY FROM PSEUDOMONAS FLUORESCENS 2-79.. ACTA CRYSTALLOGRAPHICA B. D60, 184-1862004.

Interpretive Summary: PhzF, the enzyme that catalyzes a key condensation reaction in the biosynthesis of phenazine-1-carboxylic acid in Pseudomonas fluorescens, was cloned, overexpressed, and crystallised. Data collection of native and seleno-L-methionine labelled crystals is reported.

Technical Abstract: Phenazines produced by members of several bacterial genera are biologically active metabolites that function in microbial competitiveness, the suppression of soilborne plant diseases, and virulence in infectious disease. Despite recent progress towards understanding the biochemistry of phenazine synthesis, the key reactions leading to the formation of the heterocyclic scaffold common to all phenazine compounds remain obscure. Pseudomonas fluorescens 2-79 contains seven phenazine (phz) genes that encode components of the pathway for biosynthesis of phenazine-1-carboxylic acid. A central step in this pathway involves the condensation of two identical precursor molecules derived from chorismic acid and is catalysed by the product of the phzF gene. In this study, recombinant PhzF was purified and crystallised from PEG 4000 / ammonium sulfate / sodium citrate pH 5.6. The crystals belong to spacegroup P3121 or P3221 with unit cell parameters of a=b=56.3 Å and c=156.4 Å. They contain one monomer in the asymmetric unit and diffract to better than 1.7 Å on synchrotron beamlines. Crystals of seleno-L-methionine-labelled PhzF have been obtained and SAD data are reported.