Journal
BIOTECHNOLOGY LETTERS
Volume 28, Issue 8, Pages 571-580Publisher
SPRINGER
DOI: 10.1007/s10529-006-0015-6
Keywords
amorpha-4; 11-diene synthase; Artemisia annua; enzyme expression; homologous recombination; plasmid; Saccharomyces cerevisiae; sesquiterpene production; transformation
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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 center dot 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 mu g/l of the artemisinin precursor amorpha-4,11-diene, respectively, during 16-days' batch cultivation.
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