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

Lindahl, Ann-Louise ; Olsson, Mikael E ; Mercke, Per LU ; Tollbom, Orjan ; Schelin, Jenny LU ; Brodelius, Maria and Brodelius, Peter E (2006) In Biotechnology Letters 28(8). p.80-571
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... (More)

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.

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author
; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
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
in
Biotechnology Letters
volume
28
issue
8
pages
10 pages
publisher
Springer
external identifiers
  • scopus:33645851015
  • pmid:16614895
ISSN
0141-5492
DOI
10.1007/s10529-006-0015-6
language
English
LU publication?
no
id
250aef2d-742d-45a7-a643-b031800f9a59
date added to LUP
2018-06-11 11:57:14
date last changed
2024-10-15 03:32:41
@article{250aef2d-742d-45a7-a643-b031800f9a59,
  abstract     = {{<p>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.</p>}},
  author       = {{Lindahl, Ann-Louise and Olsson, Mikael E and Mercke, Per and Tollbom, Orjan and Schelin, Jenny and Brodelius, Maria and Brodelius, Peter E}},
  issn         = {{0141-5492}},
  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}},
  language     = {{eng}},
  number       = {{8}},
  pages        = {{80--571}},
  publisher    = {{Springer}},
  series       = {{Biotechnology Letters}},
  title        = {{Production of the artemisinin precursor amorpha-4,11-diene by engineered Saccharomyces cerevisiae}},
  url          = {{http://dx.doi.org/10.1007/s10529-006-0015-6}},
  doi          = {{10.1007/s10529-006-0015-6}},
  volume       = {{28}},
  year         = {{2006}},
}