Pyruvate is a three-carbon ketoacid produced at the end of glycolysis. This research involves the metabolic engineering of the bacterium Escherichia coli in order to generate pyruvic acid (pyruvate). The Plackett-Burman design was used for a media optimization. Feeding strategies were also optimized. The addition of uncoupler 2,4-dinitrophenol increased pyruvate production and decreased lactate accumulation for CGSC6162 and CGSC6162 ppc, but the improvement is not significant. Using the optimal media for four strains CGSC6162, CGSC6162 ppc, CGSC6162 poxB, and CGSC6162 ldhA showed no significant difference in cell growth or on final pyruvate concentration. Fermentations at 37C generally produced more pyruvate and less acetate than at 40C. 37C is the optimal temperature compared to 34C and 40C.