Engineered mitochondrial production of monoterpenes in Saccharomyces cerevisiae.
Journal
Metabolic engineering
ISSN: 1096-7184
Titre abrégé: Metab Eng
Pays: Belgium
ID NLM: 9815657
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
24
04
2019
revised:
04
06
2019
accepted:
14
06
2019
pubmed:
22
6
2019
medline:
15
4
2020
entrez:
22
6
2019
Statut:
ppublish
Résumé
Monoterpene indole alkaloids (MIAs) from plants encompass a broad class of structurally complex and medicinally valuable natural products. MIAs are biologically derived from the universal precursor strictosidine. Although the strictosidine biosynthetic pathway has been identified and reconstituted, extensive work is required to optimize production of strictosidine and its precursors in yeast. In this study, we engineered a fully integrated and plasmid-free yeast strain with enhanced production of the monoterpene precursor geraniol. The geraniol biosynthetic pathway was targeted to the mitochondria to protect the GPP pool from consumption by the cytosolic ergosterol pathway. The mitochondrial geraniol producer showed a 6-fold increase in geraniol production compared to cytosolic producing strains. We further engineered the monoterpene-producing strain to synthesize the next intermediates in the strictosidine pathway: 8-hydroxygeraniol and nepetalactol. Integration of geraniol hydroxylase (G8H) from Catharanthus roseus led to essentially quantitative conversion of geraniol to 8-hydroxygeraniol at a titer of 227 mg/L in a fed-batch fermentation. Further introduction of geraniol oxidoreductase (GOR) and iridoid synthase (ISY) from C. roseus and tuning of the relative expression levels resulted in the first de novo nepetalactol production. The strategies developed in this work can facilitate future strain engineering for yeast production of later intermediates in the strictosidine biosynthetic pathway.
Identifiants
pubmed: 31226348
pii: S1096-7176(19)30182-X
doi: 10.1016/j.ymben.2019.06.004
pmc: PMC6717016
mid: NIHMS1532874
pii:
doi:
Substances chimiques
Mitochondrial Proteins
0
Monoterpenes
0
Plant Proteins
0
Vinca Alkaloids
0
strictosidine
20824-29-7
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
76-84Subventions
Organisme : NCCIH NIH HHS
ID : R01 AT010001
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM008496
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM067555
Pays : United States
Informations de copyright
Copyright © 2019 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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