Production of the artemisinin precursor amorpha-4,11-diene by engineered Saccharomyces cerevisiae

Research output: Contribution to journalArticle

Abstract

The gene encoding for amorpha-4,11-diene synthase from Artemisia annua was transformed into yeast Saccharomyces cerevisiae in two fundamentally different ways. First, the gene was subcloned into the galactose-inducible, high-copy number yeast expression vector pYeDP60 and used to transform the Saccharomyces cerevisiae strain CEN.PK113-5D. Secondly, amorpha-4,11-diene synthase gene, regulated by the same promoter, was introduced into the yeast genome by homologous recombination. In protein extracts from galactose-induced yeast cells, a higher activity was observed for yeast expressing the enzyme from the plasmid. The genome-transformed yeast grows at the same rate as wild-type yeast while plasmid-carrying yeast grows somewhat slower than the wild-type yeast. The plasmid and genome-transformed yeasts produced 600 and 100 microg/l of the artemisinin precursor amorpha-4,11-diene, respectively, during 16-days' batch cultivation.

Details

Authors
  • Ann-Louise Lindahl
  • Mikael E Olsson
  • Per Mercke
  • Orjan Tollbom
  • Jenny Schelin
  • Maria Brodelius
  • Peter E Brodelius
External organisations
  • University of Kalmar
Research areas and keywords

Keywords

  • Alkyl and Aryl Transferases/genetics, Artemisia annua/enzymology, Artemisinins/chemistry, Blotting, Western, Gas Chromatography-Mass Spectrometry, Genetic Engineering/methods, Genome, Fungal/genetics, Molecular Structure, Plasmids/genetics, Polyisoprenyl Phosphates/chemistry, Recombination, Genetic, Saccharomyces cerevisiae/genetics, Sesquiterpenes/chemistry, Transformation, Genetic
Original languageEnglish
Pages (from-to)571-80
Number of pages10
JournalBiotechnology Letters
Volume28
Issue number8
Publication statusPublished - 2006 Apr
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes