Phillips, Brian2017-09-122017-09-122017-05http://udspace.udel.edu/handle/19716/21649The metabolism of alanine (Ala), aspartate (Asp), and glutamine (Gln) in Escherichia coli was studied using 13C-metabolic flux analysis (13C-MFA) to assist future efforts to engineer methylotrophy (i.e. methanol utilization) for this organism. Growth analysis were performed using Ala, Asp, and Gln as substrates which revealed that E. coli has biomass specific substrate uptake rates [mmol/g DW biomass/hr] of 11.6, 5.93, and 1.83, respectively, for the three respective substrates. The biomass specific growth rates [1/hr] for Ala, Asp, and Gln culture were 0.222, 0.138, and 0.033, respectively. Parallel 13C tracer experiments were also performed using 13C-Ala, 13C-Asp, and 13C-Gln tracers and the labeling data was analyzed using 13C-MFA. The resulting flux results revealed that amino acid metabolism of Ala, Asp, and Gln in E. coli is characterized by high fluxes through the TCA cycle in central carbon metabolism with much lower fluxes through other metabolic pathways.Biomolecular Engineering, E. coli, amino acid metabolismThesis