Enhancement of Farnesyl Diphosphate Pool as Direct Precursor of Sesquiterpenes Through Metabolic Engineering of the Mevalonate Pathway in Saccharomyces cerevisiae
Journal article, 2010
The mevalonate pathway in the yeast Saccharomyces cerevisiae was deregulated in order to enhance the intracellular pool of farnesyl diphosphate (FPP), the direct precursor for the biosynthesis of sesquiterpenes. Overexpression of the catalytic domain of HMG1, both from the genome and plasmid, resulted in higher production of cubebol, a plant originating sesquiterpene, and increased squalene accumulation. Down-regulation of ERG9 by replacing its native promoter with the regulatable MET3 promoter, enhanced cubebol titers but simultaneous overexpression of tHMG1 and repression of ERG9 did not further improve cubebol production. Furtheremore, the concentrations of squalene and ergosterol were measured in the engineered strains. Unexpectedly, significant accumulation of squalene and restoring the ergosterol biosynthesis were observed in the ERG9 repressed strains transformed with the plasmids harboring cubebol synthase gene. This could be explained by a toxicity effect of cubebol, possibly resulting in higher transcription levels for the genes under control of MET3 promoter, which could lead to accumulation of squalene and ergosterol. Biotechnol. Bioeng. 2010;106: 86-96. (C) 2010 Wiley Periodicals, Inc.
HIGH-LEVEL PRODUCTION
YEAST SNF1
ARTEMISIA-ANNUA
CATALYTIC DOMAIN
ACTIVATED PROTEIN-KINASE
squalene
sesquiterpene
Saccharomyces cerevisiae
SQUALENE SYNTHETASE
TRANSCRIPTIONAL REGULATION
metabolic engineering
mevalonate pathway
BIOSYNTHETIC-PATHWAY
farnesyl diphosphate
SYNTHASE
HYDROXYMETHYLGLUTARYL-COA REDUCTASE
Author
M. A. Asadollahi
University of Isfahan
Danmarks Tekniske Universitet
J. Maury
Fluxome Sciences AS
Danmarks Tekniske Universitet
M. Schalk
Firmenich SA
A. Clark
Firmenich Inc.
Jens B Nielsen
Chalmers, Chemical and Biological Engineering, Life Sciences
Biotechnology and Bioengineering
0006-3592 (ISSN) 1097-0290 (eISSN)
Vol. 106 1 86-96Subject Categories (SSIF 2011)
Biological Sciences
Areas of Advance
Life Science Engineering (2010-2018)
DOI
10.1002/bit.22668