3 research outputs found
Controlling Citrate Synthase Expression by CRISPR/Cas9 Genome Editing for <i>n</i>‑Butanol Production in <i>Escherichia coli</i>
Genome
editing using CRISPR/Cas9 was successfully demonstrated in <i>Esherichia coli</i> to effectively produce <i>n</i>-butanol in a defined medium under microaerobic condition. The butanol
synthetic pathway genes including those encoding oxygen-tolerant alcohol
dehydrogenase were overexpressed in metabolically engineered <i>E. coli</i>, resulting in 0.82 g/L butanol production.
To increase butanol production, carbon flux from acetyl-CoA to citric
acid cycle should be redirected to acetoacetyl-CoA. For this purpose,
the 5′-untranslated region sequence of <i>gltA</i> encoding citrate synthase was designed using an expression prediction
program, UTR designer, and modified using the CRISPR/Cas9 genome editing
method to reduce its expression level. <i>E. coli</i> strains with decreased citrate synthase expression produced more
butanol and the citrate synthase activity was correlated with butanol
production. These results demonstrate that redistributing carbon flux
using genome editing is an efficient engineering tool for metabolite
overproduction