Metabolic engineering of Aeromonas hydrophila for the enhanced production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)

Appl Microbiol Biotechnol. 2006 Jan;69(5):537-42. doi: 10.1007/s00253-005-0034-6. Epub 2005 Jun 28.

Abstract

Wild-type Aeromonas hydrophila 4AK4 produced 35-45 wt.% poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) consisting of 10-15 mol% 3-hydroxyhexanoate (3HHx). To enhance PHBHHx production, vgb gene encoding Vitreoscilla haemoglobin or fadD gene encoding Escherichia coli acyl-CoA synthase was co-expressed with polyhydroxyalkanoates (PHA) synthesis-related genes including phbAB from Wautersia eutropha and phaPCJ from A. hydrophila. Expression of vgb increased PHBHHx content from 46 to 53 wt.% without affecting the polymer monomers composition, whereas fadD increased both PHBHHx content from 46 to 64 wt.% and its 3HHx fraction from 15 to 24 mol%. Co-expression of vgb or fadD gene with PHA-synthesis-related genes generally increased PHBHHx content over 60 wt.%. Co-expression of phbAB with vgb increased PHBHHx content and concentration up to about 70 wt.% and 4.0 g l-1, respectively. Fermentor study also showed that in the recombinants harboring vgb, CDW, PHBHHx concentration and productivity were significantly elevated up to 54 g l-1, 28.5 g l-1 and 0.791 g l-1 h-1, respectively, suggesting that vgb could promote PHA synthesis. In this strain, lac promoter could be used to constitutively express foreign genes such as phbA and phbB encoding beta-ketothiolase and NADPH-dependent acetoacetyl-CoA reductase of W. eutropha, respectively, without use of IPTG. The results showed that combined expression of different genes was a successful strategy to enhance PHA production, which could be useful for strain development to construct other recombinant PHA-producing strains.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Hydroxybutyric Acid / chemistry
  • 3-Hydroxybutyric Acid / metabolism*
  • Acetyl-CoA C-Acyltransferase / genetics
  • Aeromonas hydrophila / genetics*
  • Aeromonas hydrophila / metabolism*
  • Aldehyde Oxidoreductases / genetics
  • Bacterial Proteins / genetics
  • Caproates / chemistry
  • Caproates / metabolism*
  • Cloning, Molecular
  • Coenzyme A Ligases
  • Cupriavidus necator / genetics
  • DNA-Binding Proteins / genetics
  • Escherichia coli / genetics
  • Escherichia coli Proteins
  • Fermentation
  • Gene Expression
  • Hemeproteins / genetics
  • Lac Operon / genetics
  • Plasmids
  • Promoter Regions, Genetic
  • Recombination, Genetic
  • Vitreoscilla / genetics

Substances

  • Bacterial Proteins
  • Caproates
  • DNA-Binding Proteins
  • Escherichia coli Proteins
  • Hemeproteins
  • PHAP protein, Bacteria
  • Vgb protein, Bacteria
  • poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)
  • Aldehyde Oxidoreductases
  • acetaldehyde dehydrogenase (acylating)
  • Acetyl-CoA C-Acyltransferase
  • Coenzyme A Ligases
  • acyl-coenzyme A synthetase, E coli
  • 3-Hydroxybutyric Acid