Heterologous Production of Fosfomycin and Identification of the Minimal Biosynthetic Gene Cluster

Abstract

Fosfomycin is a clinically utilized, highly effective antibiotic, which is active against methicillin- and vancomycin-resistant pathogens. Here we report the cloning and characterization of a complete fosfomycin biosynthetic cluster from Streptomyces fradiae and heterologous production of fosfomycin in S. lividans. Sequence analysis coupled with gene deletion and disruption revealed that the minimal cluster consists of fom1-4, fomA-D. A LuxR-type activator that was apparently required for heterologous fosfomycin production was also discovered approximately 13 kb away from the cluster and was named fomR. The genes fomE and fomF, previously thought to be involved in fosfomycin biosynthesis, were shown not to be essential by gene disruption. This work provides new insights into fosfomycin biosynthesis and opens the door for fosfomycin overproduction and creation of new analogs via biomolecular pathway engineering.

Discovery of phosphonic acid natural products by mining the genomes of 10,000 actinomycetes

“…acute cystitis; FR-900098 and fosmidomycin, antimicrobials undergoing clinical trials for malaria; and phosphinothricin, the active component in several commercial herbicides (Liberty, Basta, and Rely). Second, the methodology needed for gene-based discovery of phosphonate biosynthetic loci has been rigorously established (17)(18)(19)(20)(21)(22)(23). This method relies on the fact that all but two characterized phosphonate biosynthetic pathways begin with the enzyme phosphoenolpyruvate (PEP) mutase (encoded by pepM) (14).…”

Fosfomycin and Its Application in the Treatment of Multidrug-Resistant Enterobacteriaceae Infections

“…1). Initially released by a few bacterial species, such as Streptomyces spp [26][27][28] or Pseudomonas syringae, 29 it is now produced synthetically for pharmaceutical purposes. The disodium salt is administered parenterally and the trometamine salt orally.…”

Prospecting for new bacterial metabolites: a glossary of approaches for inducing, activating and upregulating the biosynthesis of bacterial cryptic or silent natural products

“…86 In a similar way, Erwinia carotovora produces the carbapenem antibiotics (53). 87 The triggered production of other compounds including fosfomycin (54), 88 geldanamycin (55), 89 pimaricin (56 90 pikromycin (57), 91 rapamycin (6) 92 and stambomycin (58) In a recent study to unlock a cryptic biosynthetic cluster encoding novel naphthalenic octaketide ansamycins, Shen and co-workers firstly investigated the introduction of the strong consitutive promoter ermE* (in a strategy similar to that reported by Bode in 4.2), this resulted in partial activation of the pathway with an increase in the transcriptional levels of only two of the genes. Analysis of the biosynthetic cluster indicated that a putative positive regulator of the LuxR family of transcription factors was encoded, nam1.…”