A Functional Analysis of the Purine Salvage Pathway in Acetobacter fabarum

Acetobacter species are a major component of the gut microbiome of the fruit fly Drosophila melanogaster, a widely used model organism. While a range of studies have illuminated impacts of Acetobacter on their hosts, less is known about how association with the host impacts bacteria. A previous study identified that a purine salvage locus was commonly found in Acetobacter associated with Drosophila. In this study, we sought to verify the functions of predicted purine salvage genes in Acetobacter fabarum DsW_054 and to test the hypothesis that these bacteria can utilize host metabolites as a sole source of nitrogen. Targeted gene deletion and complementation experiments confirmed that genes encoding xanthine dehydrogenase (xdhB), urate hydroxylase (urhA), and allantoinase (puuE) were required for growth on their respective substrates as the sole source of nitrogen. Utilization of urate by Acetobacter is significant because this substrate is the major nitrogenous waste product of Drosophila, and its accumulation in the excretory system is detrimental to both flies and humans. The potential significance of our findings for host purine homeostasis and health are discussed, as are the implications for interactions among microbiota members, which differ in their capacity to utilize host metabolites for nitrogen. IMPORTANCEAcetobacter are commonly found in the gut microbiota of fruit flies, including Drosophila melanogaster. We evaluated the function of purine salvage genes in Acetobacter fabarum to test the hypothesis that this bacterium can utilize host metabolites as a source of nitrogen. Our results identify functions for three genes required for growth on urate, a major host waste product. The utilization of this and other Drosophila metabolites by gut bacteria may play a role in their survival in the host environment. Future research into how microbial metabolism impacts host purine homeostasis may lead to therapies because urate accumulation in the excretory system is detrimental to flies and humans.