Increase of copy number of HMG-CoA reductase and FPP synthase genes improves the amorpha4,11-diene production in engineered yeast / 药学学报
Acta Pharmaceutica Sinica
;
(12): 1314-1319, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-268185
ABSTRACT
The gene encoding amorpha-4, 11-diene synthase was cloned from Artemisia annua L. Other two genes encoding the FPP synthase (FPPS) and HMG-CoA reductase (HMGR) were cloned from Saccharomyces cerevisiae. The cloned cDNAs were confirmed by DNA sequencing. Two expression vectors were constructed, one is named pGBT9/A/HMG/FPP harboring genes for HMG-CoA reductase and FPP synthase and the other is pYeDP60/G/AS, containing the gene encoding amorpha-4,11-diene synthase. Two kinds of engineered yeast were constructed the first was named WHT [AS], which contained the plasmid pYeDP60/G/AS; the second was WHT [HMG + FPP + AS], in which the vectors pGBT9/A/ HMG/FPP and pYeDP60/G/AS were introduced by cotransformation mediated with LiOAc and PEG4000. The positive clones were identified for further fermentations. The samples from fermentations were analyzed by GC-MS for amorpha-4,11-diene. The results show that engineered yeasts could produce amorpha-4,11-diene. Moreover, the amorpha-4,11-diene production of WHT[ HMG + FPP + AS] and WHT[ AS] were 23.6 mg x L(-1) and 10 microg x L(-1), respectively. Its concentrations were reported as equivalents of valencene. The results showed the copy number increase of HMGR and FPPS genes can improve the production of amorpha-4, 11-diene in the fermentation of engineered yeasts.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Saccharomyces cerevisiae
/
Sesquiterpenes
/
Genetic Engineering
/
Genes, Plant
/
Gene Dosage
/
Alkyl and Aryl Transferases
/
Artemisia annua
/
Geranyltranstransferase
/
Fermentation
/
Genetic Vectors
Language:
Chinese
Journal:
Acta Pharmaceutica Sinica
Year:
2007
Type:
Article
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