title: Tuning primary metabolism for heterologous pathway productivity.
abstract: Tuning expression of competing endogenous pathways has been identified as an effective strategy in the optimization of heterologous production pathways. However, intervention at the first step of glycolysis, where no alternate routes of carbon utilization exist, remains unexplored. In this work we have engineered a viable E. coli host that decouples glucose transport and phosphorylation, enabling independent control of glucose flux to a heterologous pathway of interest through glucokinase (glk) expression. Using community sourced and curated promoters, glk expression was varied over a 3-fold range while maintaining cellular viability. The effects of glk expression on the productivity of a model glucose-consuming pathway were also studied. Through control of glycolytic flux we were able to explore a number of cellular phenotypes and vary the yield of our model pathway by up to 2-fold in a controllable manner.
authors: Department of Chemical Engineering, Synthetic Biology Engineering Research Center (SynBERC), Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors: Kevin V
authors: KV
authors: Solomon
authors: Tae Seok
authors: TS
authors: Moon
authors: Brian
authors: B
authors: Ma
authors: Tarielle M
authors: TM
authors: Sanders
authors: Kristala L J
authors: KL
authors: Prather
mesh_terms: D001692:Biological Transport; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D005941:Glucokinase / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000235:genetics / Q000378:metabolism*; D006019:Glycolysis; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D015398:Signal Transduction
chemical_list: D005941:Glucokinase; D005947:Glucose