Overproduction of Geranylgeraniol by Metabolically Engineered Saccharomyces cerevisiae
Open Access
- 1 September 2009
- journal article
- research article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 75 (17), 5536-5543
- https://doi.org/10.1128/aem.00277-09
Abstract
(E, E, E)-Geranylgeraniol (GGOH) is a valuable starting material for perfumes and pharmaceutical products. In the yeast Saccharomyces cerevisiae, GGOH is synthesized from the end products of the mevalonate pathway through the sequential reactions of farnesyl diphosphate synthetase (encoded by the ERG20 gene), geranylgeranyl diphosphate synthase (the BTS1 gene), and some endogenous phosphatases. We demonstrated that overexpression of the diacylglycerol diphosphate phosphatase (DPP1) gene could promote GGOH production. We also found that overexpression of a BTS1-DPP1 fusion gene was more efficient for producing GGOH than coexpression of these genes separately. Overexpression of the hydroxymethylglutaryl-coenzyme A reductase (HMG1) gene, which encodes the major rate-limiting enzyme of the mevalonate pathway, resulted in overproduction of squalene (191.9 mg liter−1) rather than GGOH (0.2 mg liter−1) in test tube cultures. Coexpression of the BTS1-DPP1 fusion gene along with the HMG1 gene partially redirected the metabolic flux from squalene to GGOH. Additional expression of a BTS1-ERG20 fusion gene resulted in an almost complete shift of the flux to GGOH production (228.8 mg liter−1 GGOH and 6.5 mg liter−1 squalene). Finally, we constructed a diploid prototrophic strain coexpressing the HMG1, BTS1-DPP1, and BTS1-ERG20 genes from multicopy integration vectors. This strain attained 3.31 g liter−1 GGOH production in a 10-liter jar fermentor with gradual feeding of a mixed glucose and ethanol solution. The use of bifunctional fusion genes such as the BTS1-DPP1 and ERG20-BTS1 genes that code sequential enzymes in the metabolic pathway was an effective method for metabolic engineering.This publication has 45 references indexed in Scilit:
- Induction of multiple pleiotropic drug resistance genes in yeast engineered to produce an increased level of anti-malarial drug precursor, artemisinic acidBMC Biotechnology, 2008
- High-Level Production of Beta-Carotene in Saccharomyces cerevisiae by Successive Transformation with Carotenogenic Genes from Xanthophyllomyces dendrorhousApplied and Environmental Microbiology, 2007
- Production of the antimalarial drug precursor artemisinic acid in engineered yeastNature, 2006
- Geranylgeraniol, an Intermediate Product in Mevalonate Pathway, Induces Apoptotic Cell Death in Human Hepatoma Cells: Death Receptor‐independent Activation of Caspase‐8 with Down‐regulation of Bcl‐xL ExpressionJapanese Journal of Cancer Research, 2001
- Construction and Evaluation of a Novel Bifunctional N -Carbamylase– d -Hydantoinase Fusion EnzymeApplied and Environmental Microbiology, 2000
- Enhanced Secretion of Human Nerve Growth Factor from Saccharomyces cerevisiae using an Advanced δ–Integration SystemBio/Technology, 1991
- Mechanism of high-copy-number integration of pMIRY-type vectors into the ribosomal DNA of Saccharomyces cerevisiaeGene, 1991
- Influence of growth rate on the accumulation of ergosterol in yeast-cellsBiotechnology Letters, 1990
- High-copy-number integration into the ribosomal DNA of Saccharomyces cerevisiae: a new vector for high-level expressionGene, 1989
- Non enzymic formation of nerolidol from farnesyl pyrophosphate in the presence of bivalent cationsBiochemical and Biophysical Research Communications, 1971