Abstract
High-throughput genetics is a powerful approach to identify new genes involved in bacterial carbon catabolism. Here, we used genome-wide fitness assays to identify a novel pathway for 2-deoxy-D-ribose catabolism in Pseudomonas simiae WCS417. The genes that were important for deoxyribose utilization but not in other conditions included two putative dehydrogenases, a lactonase, a β-keto acid cleavage enzyme, and a glycerate kinase. We propose that deoxyribose is oxidized twice to 2-deoxy-3-keto-D-ribonoate (a β-keto acid) before cleavage to D-glycerate, which is phosphorylated and enters lower glycolysis. We purified the two dehydrogenases and reconstituted the enzymatic pathway for the conversion of deoxyribose to 2-deoxy-3-keto-D-ribonoate in vitro.