Engineering a glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O2 deprivation

Biotechnol Lett. 2016 Oct;38(10):1791-7. doi: 10.1007/s10529-016-2166-4. Epub 2016 Jul 9.

Abstract

Objective: To explore the glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O2 deprivation.

Result: Overexpression of a glycerol facilitator, glycerol dehydrogenase and dihydroxyacetone kinase from Escherichia coli K-12 in C. glutamicum led to recombinant strains NC-3G diverting glycerol utilization towards succinate production under O2 deprivation. Under these conditions, strain NC-3G efficiently consumed glycerol and produced succinate without growth. The recombinant C. glutamicum utilizing glycerol as the sole carbon source showed higher intracellular NADH/NAD(+) ratio compare with utilizing glucose. The mass conversion of succinate increased from 0.64 to 0.95. Using an anaerobic fed-batch fermentation process, the final strain produced 38.4 g succinate/l with an average yield of 1.02 g/g.

Conclusions: The metabolically-engineered strains showed an efficient succinate production using glycerol as sole carbon source under O2 deprivation.

Keywords: Corynebacterium glutamicum; Glycerol utilization; NADH generation; Succinate production.

MeSH terms

  • Anaerobiosis
  • Batch Cell Culture Techniques
  • Corynebacterium glutamicum / genetics*
  • Corynebacterium glutamicum / physiology
  • Escherichia coli K12 / enzymology
  • Escherichia coli K12 / genetics
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Fermentation
  • Glycerol / metabolism*
  • Metabolic Engineering / methods
  • Phosphotransferases (Alcohol Group Acceptor) / genetics*
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Succinates / metabolism*
  • Sugar Alcohol Dehydrogenases / genetics*
  • Sugar Alcohol Dehydrogenases / metabolism

Substances

  • Escherichia coli Proteins
  • Succinates
  • Sugar Alcohol Dehydrogenases
  • glycerol dehydrogenase
  • Phosphotransferases (Alcohol Group Acceptor)
  • glycerone kinase
  • Glycerol