Characterization of a (pckA) mutant of the non-nodulating bacterium Rhizobium pusense NRCPB10 induced by transposon Tn5 mutagenesis

Phosphoenolpyruvate carboxykinase (PCK) catalyzes the decarboxylation and phosphorylation of oxaloacetate to phosphoenolpyruvate in the gluconeogenic pathway in most organisms. A pckA gene encoding PCK was cloned and sequenced from strain Rhizobium pusense NRCPB100, a spontaneous rifampicin resistant mutant of R. pusense NRCPB10(T) (JCM16209(T)) of a recently described new species of a non-nodulating and non-tumorigenic bacterium. The mapping of the pck gene region following Tn5 mutagenesis located the gene downstream of a transcriptional regulatory protein gene (chvI) and upstream of a conserved hypothetical protein gene. The pck of 1,611 bp was deduced to encode 536 amino acids and showed high homology to the genes of known ATP-dependent PCK enzymes. Phylogenetic analysis of the gene placed it in a cluster with pck of other known members of Rhizobiales. Amino acid sequences of the putative functional regions of the deduced enzyme were found to be conserved.