Recent concerns over the sustainability of petrochemical-based processes for production of desired
chemicals have fueled research into alternative modes of production. Metabolic engineering of microbial
cell factories such as Saccharomyces cerevisiae and Escherichia coli offers a sustainable and flexible
alternative for the production of various molecules. Acetyl-CoA is a key molecule in microbial central
carbon metabolism and is involved in a variety of cellular processes. In addition, it functions as a
precursor for many molecules of biotechnological relevance. Therefore, much interest exists in
engineering the metabolism around the acetyl-CoA pools in cells in order to increase product titers.
Here we provide an overview of the acetyl-CoA metabolism in eukaryotic and prokaryotic microbes
(with a focus on S. cerevisiae and E. coli), with an emphasis on reactions involved in the production and
consumption of acetyl-CoA. In addition, we review various strategies that have been used to increase
acetyl-CoA production in these microbes.