Biotechnology can serve as an attractive method to help replace products derived from nonrenewable, petroleum-derived processes with those created by microbes. One factor that must be considered when using microbes to generate a product is that a portion of the carbon supplied must be used to generate biomass. The proportion of available carbon flowing to biomass is not fixed, so reducing the fraction of carbon flowing to biomass by operational or genetic means can increase product yields. This work focuses on reducing, but not eliminating, the activity of citrate synthase to direct a greater fraction of glucose toward products derived from acetyl-CoA, while still maintaining cellular growth. The first study investigates the creation and screening of 552 citrate synthase gene mutants for increased acetate formation. A total of 16 candidate strains were identified for gene sequencing and further investigations in shake flasks; a strong negative correlation emerged between increased acetate yield and growth rate. Ultimately, 2 strains were found to produce over 6-fold more acetate than wild-type. Select strains were then chosen for batch growth studies to determine the course of acetate accumulation and to measure growth rate under controlled conditions, and the results largely paralleled those in shake flasks. Finally, chemostat experiments were performed with the wild-type strain and two strains with degraded citrate synthase activity. The most hindered strain exhibited lower biomass yield, 10-fold greater acetate yield, and 40% higher glucose uptake compared to the wild-type. The second study focuses on improving mevalonate production with wild-type E. coli, a citrate synthase deletion strain, and 9 citrate synthase variants. When grown in glucose medium supplemented with casamino acids, the wild-type strain produced the most biomass and least mevalonate, while the knockout strain demonstrated the opposite. All variant strains produced intermediate amounts of biomass and mevalonate. The wild-type, knockout strain, and three variant strains were then selected for batch growth in controlled bioreactors. A variant demonstrated the highest mevalonate volumetric productivity, 80% greater than the wild-type. This strain was selected for further process optimization using nitrogen-starved and nitrogen-limited conditions, where it ultimately produced 36.9 g/L of mevalonate in 31 h.