Nitrogen assimilation in denitrifier Bacillus azotoformans LMG 9581T.

Abstract

Until recently, it has not been generally known that some bacteria can contain the gene inventory for both denitrification and dissimilatory nitrate (NO3-)/nitrite (NO2-) reduction to ammonium (NH4+) (DNRA). Detailed studies of these microorganisms could shed light on the differentiating environmental drivers of both processes without interference of organism-specific variation. Genome analysis of Bacillus azotoformans LMG 9581T shows a remarkable redundancy of dissimilatory nitrogen reduction, with multiple copies of each denitrification gene as well as DNRA genes nrfAH, but a reduced capacity for nitrogen assimilation, with no nas operon nor amtB gene. Here, we explored nitrogen assimilation in detail using growth experiments in media with different organic and inorganic nitrogen sources at different concentrations. Monitoring of growth, NO3- NO2-, NH4+ concentration and N2O production revealed that B. azotoformans LMG 9581T could not grow with NH4+ as sole nitrogen source and confirmed the hypothesis of reduced nitrogen assimilation pathways. However, NH4+ could be assimilated and contributed up to 50% of biomass if yeast extract was also provided. NH4+ also had a significant but concentration-dependent influence on growth rate. The mechanisms behind these observations remain to be resolved but hypotheses for this deficiency in nitrogen assimilation are discussed. In addition, in all growth conditions tested a denitrification phenotype was observed, with all supplied NO3- converted to nitrous oxide (N2O).