24 Scopus citations

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.

Original languageEnglish
Pages (from-to)126-135
Number of pages10
JournalACS synthetic biology
Volume2
Issue number3
DOIs
StatePublished - Mar 15 2013

Keywords

  • flux optimization
  • glucose utilization
  • metabolic engineering
  • primary metabolism
  • tuning of gene expression

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