Graduate Thesis Or Dissertation
 

Metabolic uncoupling of Shewanella oneidensis MR-1, under the presence of excess substrate and 3,3’,4’,5 tetrachlorosalicylanilide

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  • Dissociation between catabolism and anabolism, generically termed “metabolic uncoupling”, has been studied for aerobic cultures of Shewanella oneidensis MR-1. Under conditions promoting metabolic uncoupling, the cell yield diminishes while the substrate utilization rate remains largely unchanged. The effects of excess substrate conditions and TCS addition on the metabolism of pyruvate by S. oneidensis MR-1 were examined and modeled. In the first phase, the observed cell yield (Yobs) was successfully modeled as a function of relative substrate concentration (S0/X0), under excess substrate conditions. The degree of uncoupling was determined by uncoupling coefficient (Eu). Cellular yield was shown to decrease with increasing substrate concentrations. Preliminary experiments illustrated the effectiveness of TCS in reducing the cell concentration. In the later phase, an expression is proposed to model the effect of excess substrate as well as TCS addition, simultaneously. This model was verified by using the experimental data obtained by introduction of three different TCS doses to cells grown under excess substrate conditions. The uncoupling coefficient was used to distinguish between the uncoupling effects of TCS addition and excess substrate conditions. In our experiments, it was observed that acetate accumulated as a metabolic intermediate. Average protein content of the cells was also found to increase with an increase in concentrations of either the substrate or the uncoupler. These could be the possible pathways for consumption of substrate for non-growth associated activities. Determining an optimum combination of substrate and uncoupler concentrations could be very useful in controlling the biomass growth in engineered microbial practices like wastewater treatment and bioremediation.
  • Keywords: protein, Shewanella oneidensis MR-1, Excess-substrate, yield, Metabolic Uncoupling
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