TY - JOUR
T1 - Efficient diterpene production in yeast by engineering Erg20p into a geranylgeranyl diphosphate synthase
AU - Ignea, Codruta
AU - Trikka, Fotini A.
AU - Nikolaidis, Alexandros K.
AU - Georgantea, Panagiota
AU - Ioannou, Efstathia
AU - Loupassaki, Sofia
AU - Kefalas, Panagiotis
AU - Kanellis, Angelos K.
AU - Roussis, Vassilios
AU - Makris, Antonios M.
AU - Kampranis, Sotirios C.
N1 - Copyright © 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
PY - 2015/1/1
Y1 - 2015/1/1
N2 - Terpenes have numerous applications, ranging from pharmaceuticals to fragrances and biofuels. With increasing interest in producing terpenes sustainably and economically, there has been significant progress in recent years in developing methods for their production in microorganisms. In Saccharomyces cerevisiae, production of the 20-carbon diterpenes has so far proven to be significantly less efficient than production of their 15-carbon sesquiterpene counterparts. In this report, we identify the modular structure of geranylgeranyl diphosphate synthesis in yeast to be a major limitation in diterpene yields, and we engineer the yeast farnesyl diphosphate synthase Erg20p to produce geranylgeranyl diphosphate. Using a combination of protein and genetic engineering, we achieve significant improvements in the production of sclareol and several other isoprenoids, including cis-abienol, abietadiene and β-carotene. We also report the development of yeast strains carrying the engineered Erg20p, which support efficient isoprenoid production and can be used as a dedicated chassis for diterpene production or biosynthetic pathway elucidation. The design developed here can be applied to the production of any GGPP-derived isoprenoid and is compatible with other yeast terpene production platforms.
AB - Terpenes have numerous applications, ranging from pharmaceuticals to fragrances and biofuels. With increasing interest in producing terpenes sustainably and economically, there has been significant progress in recent years in developing methods for their production in microorganisms. In Saccharomyces cerevisiae, production of the 20-carbon diterpenes has so far proven to be significantly less efficient than production of their 15-carbon sesquiterpene counterparts. In this report, we identify the modular structure of geranylgeranyl diphosphate synthesis in yeast to be a major limitation in diterpene yields, and we engineer the yeast farnesyl diphosphate synthase Erg20p to produce geranylgeranyl diphosphate. Using a combination of protein and genetic engineering, we achieve significant improvements in the production of sclareol and several other isoprenoids, including cis-abienol, abietadiene and β-carotene. We also report the development of yeast strains carrying the engineered Erg20p, which support efficient isoprenoid production and can be used as a dedicated chassis for diterpene production or biosynthetic pathway elucidation. The design developed here can be applied to the production of any GGPP-derived isoprenoid and is compatible with other yeast terpene production platforms.
KW - Diterpenes
KW - Geranyltranstransferase
KW - Metabolic Engineering
KW - Polyisoprenyl Phosphates
KW - Saccharomyces cerevisiae
KW - Saccharomyces cerevisiae Proteins
U2 - 10.1016/j.ymben.2014.10.008
DO - 10.1016/j.ymben.2014.10.008
M3 - Journal article
C2 - 25446975
SN - 1096-7176
VL - 27
SP - 65
EP - 75
JO - Metabolic Engineering
JF - Metabolic Engineering
ER -