Increased riboflavin production by knockout of 6-phosphofructokinase I and blocking the Entner-Doudoroff pathway in Escherichia coli

Biotechnol Lett. 2016 Aug;38(8):1307-14. doi: 10.1007/s10529-016-2104-5. Epub 2016 Apr 12.

Abstract

Objectives: To construct an Escherichia coli strain capable of producing riboflavin with high titer and yield.

Results: A low copy number plasmid pLS01 containing a riboflavin operon under the control of a constitutive promoter was constructed and introduced into Escherichia coli MG1655. Subsequently, the pfkA, edd and ead genes were disrupted, and the resulting strain LS02T produced 667 mg riboflavin/l in MSY medium supplied with 10 g glucose/l in flask cultivation. In a fed-batch process, riboflavin production of the strain reached 10.4 g/l with a yield of 56.8 mg riboflavin/g glucose.

Conclusion: To our knowledge, this is the first report of engineered E. coli strains that can produce more than 10 g riboflavin/l in fed-batch cultivation, indicating that E. coli has potential for riboflavin production.

Keywords: Escherichia coli; Fed-batch fermentation; Metabolic engineering; Riboflavin.

MeSH terms

  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Fermentation / genetics
  • Fermentation / physiology
  • Industrial Microbiology
  • Phosphofructokinases / genetics*
  • Phosphofructokinases / metabolism*
  • Riboflavin / biosynthesis*
  • Riboflavin / genetics

Substances

  • Escherichia coli Proteins
  • Phosphofructokinases
  • Riboflavin