Peracchi, A. | Search bioRxiv

bioRxiv2026

Human ASPDH is a 2-aminomuconate reductase that produces L-2-aminohex-3-enedioic acid in tryptophan catabolism

Most tryptophan catabolism in animals occurs through the kynurenine pathway, which generates the essential NAD cofactor and multiple bioactive metabolites. Knowledge of this pathway in eukaryotes ends at the unstable intermediate 2-aminomuconate (2-AM). Here, by leveraging evolutionary information from more than 5,000 eukaryotes, we identify two distinct genes acting downstream of 2-AM in fungi and metazoa. The fungal gene is homologous to bacterial 2-AM deaminase, whereas the metazoan gene is homologous to aspartate dehydrogenase (ASPDH), which in prokaryotes catalyses the first reaction of NAD biosynthesis. Biochemical and structural analyses show that human ASPDH has evolved an unprecedented function as an NAD(P)H-dependent 2-AM reductase (AMR) in tryptophan catabolism. The reaction forms L-2-aminohex-3-enedioic acid, an unsaturated -amino acid absent from current biological databases. Isotope-labeling NMR experiments and structural modelling support a mechanism in which hydride transfer is coupled to double-bond rearrangement of the conjugated system. These findings reveal a previously unknown metazoan branch of the kynurenine pathway, expand the repertoire of endogenous amino acids, and illustrate how comparative genomics can uncover hidden reactions in human metabolism.

biochemistry↗

bioRxiv2020

(R)-1-hydroxy-2-aminoethylphosphonate ammonia-lyase, a new PLP-dependent enzyme involved in bacterial phosphonate degradation

Phosphonates contain a particularly stable carbon-phosphorus bond, yet a number of microorganisms possess pathways to degrade these molecules and use them as source of phosphorus. One example is the widespread hydrolytic route for the breakdown of 2-aminoethylphosphonate (AEP). In this pathway, the aminotransferase PhnW initially converts AEP into phosphonoacetaldehyde (PAA), which is then cleaved by the hydrolase PhnX to yield acetaldehyde and phosphate. This work focuses on a novel enzyme (hereby termed PbfA), which is often encoded in bacterial gene clusters containing the phnW-phnX combination. Although PbfA is annotated as a transaminase, we report that it catalyzes an elimination reaction on the naturally occurring compound (R)-1-hydroxy-2-aminoethylphosphonate (R-HAEP). The reaction releases ammonia and generates PAA, which can be subsequently hydrolyzed by PhnX. Overall, the PbfA reaction represents a frequent accessory branch in the hydrolytic pathway for AEP degradation, which expands the scope and versatility of the pathway itself.

biochemistry↗