TY - JOUR
T1 - Reconstructing the chemical diversity of labdane-type diterpene biosynthesis in yeast
AU - Ignea, Codruta
AU - Ioannou, Efstathia
AU - Georgantea, Panagiota
AU - Loupassaki, Sofia
AU - Trikka, Fotini A.
AU - Kanellis, Angelos K.
AU - Makris, Antonios M.
AU - Roussis, Vassilios
AU - Kampranis, Sotirios C.
N1 - Copyright © 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
PY - 2015/3/1
Y1 - 2015/3/1
N2 - Terpenes are a large class of natural products, many of which are used in cosmetics, pharmaceuticals, or biofuels. However, terpene[U+05F3]s industrial application is frequently hindered by limited availability of natural sources or low yields of chemical synthesis. In this report, we developed a modular platform based on standardized and exchangeable parts to reproduce and potentially expand the diversity of terpene structures in Saccharomyces cerevisiae. By combining different module-specific parts, we exploited the substrate promiscuity of class I diterpene synthases to produce an array of labdane-type scaffolds. These were subsequently modified by a scaffold decoration module consisting of a mutant library of a promiscuous cytochrome P450 to afford a range of hydroxylated diterpenes. Further P450 protein engineering yielded dedicated and efficient catalysts for specific products. Terpenes produced include precursors of pharmacologically important compounds, molecules that are difficult to obtain from natural sources, or new natural products. The approach described here provides a platform on which additional gene mining, combinatorial biosynthesis, and protein engineering efforts can be integrated to sustainably explore the terpene chemical space.
AB - Terpenes are a large class of natural products, many of which are used in cosmetics, pharmaceuticals, or biofuels. However, terpene[U+05F3]s industrial application is frequently hindered by limited availability of natural sources or low yields of chemical synthesis. In this report, we developed a modular platform based on standardized and exchangeable parts to reproduce and potentially expand the diversity of terpene structures in Saccharomyces cerevisiae. By combining different module-specific parts, we exploited the substrate promiscuity of class I diterpene synthases to produce an array of labdane-type scaffolds. These were subsequently modified by a scaffold decoration module consisting of a mutant library of a promiscuous cytochrome P450 to afford a range of hydroxylated diterpenes. Further P450 protein engineering yielded dedicated and efficient catalysts for specific products. Terpenes produced include precursors of pharmacologically important compounds, molecules that are difficult to obtain from natural sources, or new natural products. The approach described here provides a platform on which additional gene mining, combinatorial biosynthesis, and protein engineering efforts can be integrated to sustainably explore the terpene chemical space.
KW - Cytochrome P-450 Enzyme System
KW - Diterpenes
KW - Hydroxylation
KW - Saccharomyces cerevisiae
KW - Saccharomyces cerevisiae Proteins
U2 - 10.1016/j.ymben.2014.12.001
DO - 10.1016/j.ymben.2014.12.001
M3 - Journal article
C2 - 25498547
SN - 1096-7176
VL - 28
SP - 91
EP - 103
JO - Metabolic Engineering
JF - Metabolic Engineering
ER -