Denitrifying bacteria that are switched from oxic to anoxic conditions can experience diauxic lag, which is the time required for re-synthesis of nitrate reductase and other denitrifying enzymes. Pseudomonas denitrificans were exposed to alternating oxic/anoxic phases in a continuous flow reactor with either 4-h or 8-h anoxic phase lengths, in comparison to a measured diauxic lag of 9.5 h following steady-state oxic conditions. The P. denitrificans were unable to sustain anoxic growth at either of the anoxic phase lengths tested. Diauxic lag observed after several cycles of alternating oxic/anoxic phases was significantly longer than the diauxic lag measured after steady-state oxic conditions. This may be attributed to increase of cell maintenance energy requirements due to substrate accumulation during anoxic phases and concomitant high specific growth rates during oxic phases.