Multienzyme Biosynthesis of Dihydroartemisinic Acid

Molecules. 2017 Aug 28;22(9):1422. doi: 10.3390/molecules22091422.


One-pot multienzyme biosynthesis is an attractive method for producing complex, chiral bioactive compounds. It is advantageous over step-by-step synthesis, as it simplifies the process, reduces costs and often leads to higher yield due to the synergistic effects of enzymatic reactions. In this study, dihydroartemisinic acid (DHAA) pathway enzymes were overexpressed in Saccharomyces cerevisiae, and whole-cell biotransformation of amorpha-4,11-diene (AD) to DHAA was demonstrated. The first oxidation step by cytochrome P450 (CYP71AV1) is the main rate-limiting step, and a series of N-terminal truncation and transcriptional tuning improved the enzymatic activity. With the co-expression of artemisinic aldehyde dehydrogenase (ALDH1), which recycles NADPH, a significant 8-fold enhancement of DHAA production was observed. Subsequently, abiotic conditions were optimized to further enhance the productivity of the whole-cell biocatalysts. Collectively, approximately 230 mg/L DHAA was produced by the multi-step whole-cell reaction, a ~50% conversion from AD. This study illustrates the feasibility of producing bioactive compounds by in vitro one-pot multienzyme reactions.

Keywords: CYP71AV1; dihydroartemisinic acid; whole cell biocatalysis.

MeSH terms

  • Aldehyde Dehydrogenase 1
  • Artemisinins / chemistry
  • Artemisinins / metabolism*
  • Biocatalysis
  • Biosynthetic Pathways
  • Cytochrome P-450 Enzyme System / metabolism
  • Fungal Proteins / metabolism*
  • Isoenzymes / metabolism
  • Oxidation-Reduction
  • Polycyclic Sesquiterpenes
  • Retinal Dehydrogenase / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development*
  • Sesquiterpenes / metabolism


  • Aldehyde Dehydrogenase 1
  • Artemisinins
  • Fungal Proteins
  • Isoenzymes
  • Polycyclic Sesquiterpenes
  • Sesquiterpenes
  • amorpha-4,11-diene
  • dihydroartemisinic acid
  • Cytochrome P-450 Enzyme System
  • Retinal Dehydrogenase