Genetic linkage in Pseudomonas aeruginosa of algT and nadB: Mutation in nadB does not affect NAD biosynthesis or alginate production

Abstract

The 68-min region of the chromosome of Pseudomonas aeruginosa ( Pa) contains the gene algT, encoding a putative alternate sigma factor similar to σ E in Escherichia coli, that is required for the expression of several genes in the alginate biosynthetic regulon. Sequences immediately upstream from algT were found to contain a divergently expressed open reading frame encoding a 60-kDa protein with 64 and 36% identity to the nadB gene products of E. coli and Bacillus subtilis, respectively. The nadB gene encodes L-aspartate oxidase and has been shown in several bacteria to be essential for de novo nicotinamide-adenine dinucleotide (NAD) biosynthesis. Pa nadB complemented the growth requirement for nicotinic acid in a nadB mutant strain of E. coli, suggesting that this gene encodes a functional homologue of l-aspartate oxidase. A nadB::Tn 501 mutant was constructed by gene replacement in the alginate-producing strain, Pa FRD. This NadB − mutant still produced alginate and appeared normal with respect to the regulation of alginate synthesis. Interestingly, the NadB − mutant did not have an auxotrophic phenotype for nicotinic acid, indicating that this nadB was not essential for NAD biosynthesis in Pa. These results suggest the possibility that Pa has an alternate mechanism for de novo NAD biosynthesis.