Enhanced butanol production by increasing NADH and ATP levels in Clostridium beijerinckii NCIMB 8052 by insertional inactivation of Cbei_4110

Appl Microbiol Biotechnol. 2016 Jun;100(11):4985-96. doi: 10.1007/s00253-016-7299-9. Epub 2016 Feb 1.

Abstract

Clostridium beijerinckii is identified as a promising Clostridium strain for industrialization of acetone and butanol (AB) fermentation. It has been reported that high reducing power levels are associated with high butanol yield. In this study, we regulated reducing power by blocking NAD(P)H consumption in C. beijerinckii NCIMB 8052. Gene Cbei_4110, encoding NADH-quinone oxidoreductase (nuoG), is a subunit of the electron transport chain complex I. After inactivation of gene Cbei_4110, the generated mutant strain exhibited a remarkable increase in glucose utilization ratio and enhanced butanol production to 9.5 g/L in P2 medium containing 30 g/L of glucose. NAD(P)H and ATP levels were also increased by one to two times and three to five times, respectively. Furthermore, a comparative transcriptome analysis was carried out in order to determine the mechanism involved in the enhanced activity of the Cbei_4110-inactivated mutant strain. This strategy may be extended for making industrial bio-butanol more economically attractive.

Keywords: AB fermentation; ATP; C. beijerinckii; NAD(P)H; hydA; nuoG.

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Butanols / metabolism*
  • Clostridium beijerinckii / genetics*
  • Clostridium beijerinckii / metabolism
  • Culture Media / chemistry
  • Electron Transport Complex I / genetics*
  • Fermentation
  • Gene Expression Regulation, Bacterial*
  • Genes, Bacterial
  • Industrial Microbiology
  • Mutagenesis, Insertional
  • NAD / metabolism
  • NADP / metabolism
  • Plasmids / genetics
  • Plasmids / metabolism
  • Sequence Alignment
  • Transcriptome

Substances

  • Butanols
  • Culture Media
  • NAD
  • NADP
  • Adenosine Triphosphate
  • Electron Transport Complex I