Biosynthesis of fosfomycin in pseudomonads reveals an unexpected enzymatic activity in the metallohydrolase superfamily

Significance Fosfomycin is an antibiotic used for the treatment of cystitis. Its activity against both gram-positive and gram-negative pathogens has received strong recent interest. The compound is biosynthesized by both pseudomonads and streptomycetes via two different pathways that converge in the last step to form the natural product. This study investigated the biosynthesis of fosfomycin by Pseudomonas and revealed an oxidative decarboxylation activity within this pathway. Structural and sequence analysis of the enzyme identified key residues important for its activity and demonstrated its role in the biosynthesis of other phosphonate natural products. This newly characterized protein family expands the chemical space of catalysis by metallohydrolase superfamily enzymes.