Poly(3-hydroxybutyrate) synthesis by recombinant Escherichia coli arcA mutants in microaerobiosis

Appl Environ Microbiol. 2006 Apr;72(4):2614-20. doi: 10.1128/AEM.72.4.2614-2620.2006.

Abstract

We assessed the effects of different arcA mutations on poly(3-hydroxybutyrate) (PHB) synthesis in recombinant Escherichia coli strains carrying the pha synthesis genes from Azotobacter sp. strain FA8. The arcA mutations used were an internal deletion and the arcA2 allele, a leaky mutation for some of the characteristics of the Arc phenotype which confers high respiratory capacity. PHB synthesis was not detected in the wild-type strain in shaken flask cultures under low-oxygen conditions, while ArcA mutants gave rise to polymer accumulation of up to 24% of their cell dry weight. When grown under microaerobic conditions in a bioreactor, the arcA deletion mutant reached a PHB content of 27% +/- 2%. Under the same conditions, higher biomass and PHB concentrations were observed for the strain bearing the arcA2 allele, resulting in a PHB content of 35% +/- 3%. This strain grew in a simple medium at a specific growth rate of 0.69 +/- 0.07 h(-1), whereas the deletion mutant needed several nutritional additives and showed a specific growth rate of 0.56 +/- 0.06 h(-1). The results presented here suggest that arcA mutations could play a role in heterologous PHB synthesis in microaerobiosis.

MeSH terms

  • Aerobiosis
  • Bacterial Outer Membrane Proteins / genetics*
  • Biomass
  • Culture Media
  • Escherichia coli / genetics
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / genetics*
  • Escherichia coli Proteins / metabolism
  • Gene Expression Regulation, Bacterial
  • Hydroxybutyrates / metabolism*
  • Mutation*
  • Polyesters / metabolism*
  • Recombination, Genetic*
  • Repressor Proteins / genetics*

Substances

  • Bacterial Outer Membrane Proteins
  • Culture Media
  • Escherichia coli Proteins
  • Hydroxybutyrates
  • Polyesters
  • Repressor Proteins
  • arcA protein, E coli
  • poly-beta-hydroxybutyrate