Production of the artemisinin precursor amorpha-4,11-diene by engineered Saccharomyces cerevisiae.
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.
Lindahl
Ann-Louise
AL
Department of Chemistry and Biomedical Science, University of Kalmar, SE-39182, Kalmar, Sweden.
Olsson
Mikael E
ME
Mercke
Per
P
Tollbom
Orjan
O
Schelin
Jenny
J
Brodelius
Maria
M
Brodelius
Peter E
PE
D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D031172:Artemisia annua / Q000201:enzymology* / Q000235:genetics; D037621:Artemisinins / Q000737:chemistry / Q000378:metabolism*; D015153:Blotting, Western; D008401:Gas Chromatography-Mass Spectrometry; D005818:Genetic Engineering / Q000379:methods*; D016681:Genome, Fungal / Q000235:genetics; D015394:Molecular Structure; D010957:Plasmids / Q000235:genetics; D011106:Polyisoprenyl Phosphates / Q000737:chemistry / Q000378:metabolism; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism*; D014170:Transformation, Genetic
D037621:Artemisinins; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C004808:farnesyl pyrophosphate; C031327:artemisinin; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase