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Per Mercke. Fotograf Johan Persson.

Per Mercke

Patent advisor

Per Mercke. Fotograf Johan Persson.

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

Author

  • Ann-Louise Lindahl
  • Mikael E Olsson
  • Per Mercke
  • Orjan Tollbom
  • Jenny Schelin
  • Maria Brodelius
  • Peter E Brodelius

Summary, in English

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.

Publishing year

2006-04

Language

English

Pages

80-571

Publication/Series

Biotechnology Letters

Volume

28

Issue

8

Document type

Journal article

Publisher

Springer

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

Status

Published

ISBN/ISSN/Other

  • ISSN: 0141-5492