Production of D-(--)-3-hydroxyalkanoic acid by recombinant Escherichia coli

FEMS Microbiol Lett. 2003 Jan 21;218(1):59-64. doi: 10.1111/j.1574-6968.2003.tb11498.x.

Abstract

Pathways for extracellular production of chiral D-(-)-3-hydroxybutyric acid (3HB) and D-(-)-3-hydroxyalkanoic acid (mcl-3HA) were constructed by co-expression of genes of beta-ketothiolase (phbA), acetoacetyl-CoA reductase (phbB) and 3-hydroxyacyl-ACP CoA transacylase (phaG), respectively, in Escherichia coli strain DH5alpha. The effect of acrylic acid and glucose on production of both 3HB and mcl-3HA was investigated. It was found that the addition of acrylic acid significantly increased production of 3HB and mcl-3HA consisting of 3-hydroxyoctanoic acid and 3-hydroxydecanoic acid in a ratio of 1:3 from 199 mg x l(-1) to 661 mg x l(-1) and from 27 mg x l(-1) to 135 mg x l(-1), respectively, in shake flask studies when glucose was present in the medium at the very beginning of fermentation. The timing of glucose addition had no effect on 3HB production. In contrast, mcl-3HA production was affected by glucose addition, an mcl-3HA concentration of 193 mg x l(-1) was obtained when glucose was added to the culture at 12 h. A more than seven-fold increase was obtained when compared with that in medium containing glucose at the beginning of fermentation. However, a decrease in production of 3HB and mcl-3HA was found when glucose was added at 12 h to the culture containing acrylic acid. The repressive effect of acrylic acid on acetic acid production was also evaluated and discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Hydroxybutyric Acid / metabolism*
  • Acetyl-CoA C-Acyltransferase / genetics
  • Acetyl-CoA C-Acyltransferase / metabolism
  • Acyltransferases / genetics
  • Acyltransferases / metabolism
  • Alcohol Oxidoreductases / genetics
  • Alcohol Oxidoreductases / metabolism
  • Caprylates / metabolism*
  • Decanoic Acids / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Plasmids
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Caprylates
  • Decanoic Acids
  • Recombinant Proteins
  • 3-hydroxyoctanoic acid
  • Alcohol Oxidoreductases
  • acetoacetyl-CoA reductase
  • Acyltransferases
  • 3-hydroxyacyl-CoA-acyl carrier protein transferase
  • Acetyl-CoA C-Acyltransferase
  • myrmicacin
  • 3-Hydroxybutyric Acid