(228ap) Controlling Glycolytic Flux Regulation for Production of N-Acetylglucosamine in Saccharomyces Cerevisiae

We previously demonstrated that the production of N-acetylglucosamine (GlcNAc) in Saccharomyces cerevisiae was enabled by overexpressing an allosteric regulation-free GFA1 mutant and the haloacid dehalogenase-like phosphatase YqaB. Wefurther improved GlcNAc production with controlling glycolytic flux. Rructose 2,6-bisphosphate (F26BP) was known as a modulator for the activity of phosphofructokinase 1 (PFK-1), the rate limiting enzyme of glycolysis. Disruption of PFK-2 was attempted to abolish synthesis of F26BP and to increase GlcNAc production by reducing glycolytic flux, but no significant change of GlcNAc with glucose as a sole carbon source. However, when galactose was used as a sole carbon source, GlcNAc production was significantly increased in the strain without PFK-2, suggesting significant reduction of glycolytic flux is needed for improving GlcNAc production. Controlling glycolytic flux of S. cerevisiae can be applied to improve production of carbohydrate derivatives.