Large-scale genomic sequencing efforts have served to accelerate the discovery and characterization of secondary metabolite biosynthetic pathways in pathogenic bacteria. The production of many of these secondary metabolites can be influential to the virulence of the pathogen, and thus the enzymes that biosynthesize them represent potential targets of antibacterial intervention. However, as is observed for a plethora of organisms considered innocuous, the secondary metabolites themselves, or derivatives therein, may be useful molecules from an industrial perspective. The AsbF enzyme, a dehydroshikimate dehydratase, catalyzes a key step in the synthesis of the stealth siderophore petrobactin by pathogenic members of the Bacillus cereus sensu lato group. As strains deficient in petrobactin production are not able to attain virulence in a murine model, small molecule inhibition of the biosynthetic enzymes potentially represent attractive anti-Bacillus therapies. Inhibitor studies with purified AsbF were conducted in vitro using a UV kinetic assay, and have uncovered several molecular features important for binding by the enzyme. AsbF has also proven useful in the heterologous biosynthesis of the polymer precursor, β-carboxy-cis,cis-muconic acid. The target molecule was produced using a de novo biosynthetic pathway expressed from a multi-monocistronic construct in an E. coli host. Using this pathway, β-carboxy-cis,cis-muconic acid is produced from glucose after conversion to dehydroshikimic acid via the endogenous shikimate pathway. Furthermore, facile conversion of the triacid to 3-carboxymuconolactone, itself a promising industrial compound, is readily achievable upon treatment with acid.

Dehydroshikimate dehydratase from pathogenic Bacillus strains: Antibiotic target and catalyst for heterologous biosynthesis of polymer precursors

249th National Meeting (2015)

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