Picolinic acid (PA)， a typical C-2-carboxylated pyridine derivative， is a metabolite of l-tryptophan and many other aromatic compounds in mammalian and microbial cells. Microorganisms can degrade and utilize PA for growth. However， the precise mechanism of PA metabolism remains unknown. strain JQ135 utilizes PA as its carbon and nitrogen source for growth. In this study， we screened a 6-hydroxypicolinic acid (6HPA) degradation-deficient mutant through random transposon mutagenesis. The mutant hydroxylated 6HPA into an intermediate， identified as 3，6-dihydroxypicolinic acid (3，6DHPA)， with no further degradation. A novel decarboxylase， PicC， was identified to be responsible for the decarboxy... More
Picolinic acid (PA)， a typical C-2-carboxylated pyridine derivative， is a metabolite of l-tryptophan and many other aromatic compounds in mammalian and microbial cells. Microorganisms can degrade and utilize PA for growth. However， the precise mechanism of PA metabolism remains unknown. strain JQ135 utilizes PA as its carbon and nitrogen source for growth. In this study， we screened a 6-hydroxypicolinic acid (6HPA) degradation-deficient mutant through random transposon mutagenesis. The mutant hydroxylated 6HPA into an intermediate， identified as 3，6-dihydroxypicolinic acid (3，6DHPA)， with no further degradation. A novel decarboxylase， PicC， was identified to be responsible for the decarboxylation of 3，6DHPA to 2，5-dihydroxypyridine. Although， PicC belonged to the amidohydrolase 2 family， it shows low similarity (<45%) compared to other reported amidohydrolase 2 family decarboxylases. Moreover， PicC was found to form a monophyletic group in the phylogenetic tree constructed using PicC and related proteins. Further， the genetic deletion and complementation results demonstrated that was essential for PA degradation. The PicC was Zn-dependent nonoxidative decarboxylase that can specifically catalyze the irreversible decarboxylation of 3，6DHPA to 2，5-dihydroxypyridine. The and toward 3，6DHPA were observed to be 13.44 μM and 4.77 s， respectively. Site-directed mutagenesis showed that His163 and His216 were essential for PicC activity. This study provides new insights into the microbial metabolism of PA at molecular level. Picolinic acid is a natural toxic pyridine derived from l-tryptophan metabolism and other aromatic compounds in mammalian and microbial cells. Microorganisms can degrade and utilize picolinic acid for their growth， and thus a microbial degradation pathway of picolinic acid has been proposed. Picolinic acid is converted into 6-hydroxypicolinic acid， 3，6-dihydroxypicolinic acid， and 2，5-dihydroxypyridine in turn. However， there was no physiological and genetic validation for this pathway. This study demonstrated that 3，6-dihydroxypicolinic acid was an intermediate in picolinic acid catabolism and further identified and characterized a novel amidohydrolase 2 family decarboxylase PicC. PicC was also shown to catalyze the decarboxylation of 3，6-dihydroxypicolinic acid into 2，5-dihydroxypyridine. This study provides a basis for understanding picolinic acid degradation and its underlying molecular mechanism.
