5U58

Psf4 in complex with Fe2+ and (R)-2-HPP

Summary for 5U58

• Entry DOI: 10.2210/pdb5u58/pdb
• Related: 5U55 5U57 5U5D
• Descriptor: (S)-2-hydroxypropylphosphonic acid epoxidase, FE (III) ION, [(2R)-2-hydroxypropyl]phosphonic acid, ... (4 entities in total)
• Functional Keywords: fosfomycin, epoxidase, oxidase, oxidoreductase
• Biological source: Pseudomonas syringae
• Total number of polymer chains: 4
• Total formula weight: 85831.21

Authors

Chekan, J.R.,Nair, S.K. (deposition date: 2016-12-06, release date: 2017-01-04, Last modification date: 2024-03-06)

Primary citation

Olivares, P.,Ulrich, E.C.,Chekan, J.R.,van der Donk, W.A.,Nair, S.K.
Characterization of Two Late-Stage Enzymes Involved in Fosfomycin Biosynthesis in Pseudomonads.
ACS Chem. Biol., 12:456-463, 2017

PubMed Abstract: The broad-spectrum phosphonate antibiotic fosfomycin is currently in use for clinical treatment of infections caused by both Gram-positive and Gram-negative uropathogens. The antibiotic is biosynthesized by various streptomycetes, as well as by pseudomonads. Notably, the biosynthetic strategies used by the two genera share only two steps: the first step in which primary metabolite phosphoenolpyruvate (PEP) is converted to phosphonopyruvate (PnPy) and the terminal step in which 2-hydroxypropylphosphonate (2-HPP) is converted to fosfomycin. Otherwise, distinct enzymatic paths are employed. Here, we biochemically confirm the last two steps in the fosfomycin biosynthetic pathway of Pseudomonas syringae PB-5123, showing that Psf3 performs the reduction of 2-oxopropylphosphonate (2-OPP) to (S)-2-HPP, followed by the Psf4-catalyzed epoxidation of (S)-2-HPP to fosfomycin. Psf4 can also accept (R)-2-HPP as a substrate but instead performs an oxidation to make 2-OPP. We show that the combined activities of Psf3 and Psf4 can be used to convert racemic 2-HPP to fosfomycin in an enantioconvergent process. X-ray structures of each enzyme with bound substrates provide insights into the stereospecificity of each conversion. These studies shed light on the reaction mechanisms of the two terminal enzymes in a distinct pathway employed by pseudomonads for the production of a potent antimicrobial agent.

PubMed: 27977135
DOI: 10.1021/acschembio.6b00939

Experimental method

X-RAY DIFFRACTION (2.7 Å)