Characterization of a novel β-alanine biosynthetic pathway consisting of promiscuous metabolic enzymes

• Published in: The Journal of biological chemistry 2022-07, Vol.298 (7), p.102067-102067, Article 102067
• Main Authors: Perchat, Nadia, Dubois, Christelle, Mor-Gautier, Rémi, Duquesne, Sophie, Lechaplais, Christophe, Roche, David, Fouteau, Stéphanie, Darii, Ekaterina, Perret, Alain
• Format: Article
• Language: English
• Subjects: Biochemistry, Molecular Biology; CoA; enzyme kinetics; gene knockout; genomics; Life Sciences; metabolism; metabolomics; polyamine; β-alanine
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/j.jbc.2022.102067

Bacteria adapt to utilize the nutrients available in their environment through a sophisticated metabolic system composed of highly specialized enzymes. Although these enzymes can metabolize molecules other than those for which they evolved, their efficiency toward promiscuous substrates is considered too low to be of physiological relevance. Herein, we investigated the possibility that these promiscuous enzymes are actually efficient enough at metabolizing secondary substrates to modify the phenotype of the cell. For example, in the bacterium Acinetobacter baylyi ADP1 (ADP1), panD (coding for l-aspartate decarboxylase) encodes the only protein known to catalyze the synthesis of β-alanine, an obligate intermediate in CoA synthesis. However, we show that the ADP1 ΔpanD mutant could also form this molecule through an unknown metabolic pathway arising from promiscuous enzymes and grow as efficiently as the wildtype strain. Using metabolomic analyses, we identified 1,3-diaminopropane and 3-aminopropanal as intermediates in this novel pathway. We also conducted activity screening and enzyme kinetics to elucidate candidate enzymes involved in this pathway, including 2,4-diaminobutyrate aminotransferase (Dat) and 2,4-diaminobutyrate decarboxylase (Ddc) and validated this pathway in vivo by analyzing the phenotype of mutant bacterial strains. Finally, we experimentally demonstrate that this novel metabolic route is not restricted to ADP1. We propose that the occurrence of conserved genes in hundreds of genomes across many phyla suggests that this previously undescribed pathway is widespread in prokaryotes.