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

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.

• Författare: Ann-Louise Lindahl; Mikael E Olsson; Per Mercke; Orjan Tollbom; Jenny Schelin; Maria Brodelius; Peter E Brodelius
• Publiceringsår: 2006-04
• Språk: Engelska
• Sidor: 80-571
• Publikation/Tidskrift/Serie: Biotechnology Letters
• Volym: 28
• Issue: 8
• Länkar: http://dx.doi.org/10.1007/s10529-006-0015-6
• Dokumenttyp: Artikel i tidskrift
• Förlag: Springer
• Nyckelord: 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
• Aktiv: Published
• ISBN/ISSN/Övrigt: ISSN: 0141-5492