Dynamics of Substrate Consumption and Enzyme Synthesis in Chelatobacter heintzii during Growth in Carbon-Limited Continuous Culture with Different Mixtures of Glucose and Nitrilotriacetate

Abstract

Regulation of nitrilotriacetate (NTA) degradation and expression of NTA monooxygenase (NTA-MO) in the NTA-degrading strain Chelatobacter heintzii ATCC 29600 in continuous culture at a dilution rate of 0.06 h(sup-1) under transient growth conditions when the feed was switched between media containing NTA, glucose, or different mixtures thereof as the sole carbon and energy sources was investigated. A transition from NTA to glucose was accompanied by a rapid loss of NTA-MO. A transition from glucose to NTA resulted in a lag phase of some 25 h until NTA-MO expression started, and approximately 100 h was needed before a steady state for NTA-MO specific activity was reached. This transient lag phase was markedly shortened when mixtures of NTA plus glucose were supplied instead of NTA only; for example, when a mixture of 90% glucose and 10% NTA was used, induction of NTA-MO was detected after 30 min. This suggests a strong positive influence of alternative carbon substrates on the expression of other enzymes under natural environmental conditions. Regulation of NTA-MO expression and the fate of NTA-MO were also studied during starvation of both glucose-grown and NTA-grown cultures. Starvation of NTA-grown cells led to a loss of NTA-MO protein. No synthesis of NTA-MO (derepression) was observed when glucose-grown cells were starved.