Global Gene Expression in Bradyrhizobium japonicum Cultured with Vanillin, Vanillate, 4-Hydroxybenzoate and Protocatechuate

Abstract

Pathways for aerobic degradation of naturally occurring aromatics were estimated from the entire genome sequence of Bradyrhizobium japonicum strain USDA110, a symbiotic nitrogen-fixing bacterium in soil. Many homologs for the genes encoding various oxygenases and enzymes for the β-ketoadipate pathway in the degradation of vanillin, vanillate, protocatechuate, and 4-hydroxybenzoate were scattered over nine loci of the genome. Using a macroarray developed for B. japonicum strain USDA110, we compared gene expression profiles in cells grown in each of these aromatic compounds as a sole carbon source with those of succinate-fed cells. One set of oxygenase genes homologous to pcaGH, pobA, and vanAB and structurally accompanied by transcriptional regulator homologs was markedly upregulated in their expression by one or more of the four aromatics, whereas no marked change was observed in the expression levels of pcaBCDIJF genes for the β-ketoadipate pathway. In addition, cells fed vanillin and vanillate showed high levels of expression of genes for a glutathione-dependent pathway of formaldehyde oxidation, suggesting that the formaldehyde generated from vanillate's demethylation is oxidized via C1 metabolism in B. japonicum. The expression of the above genes was confirmed by quantitative reverse transcription PCR. The implications of these results are discussed in terms of degradation pathways, gene regulation, and the soil environment.