Metabolome and proteome analyses reveal transcriptional misregulation in glycolysis of engineered E. coli.
Algorithms
Bacterial Proteins
/ genetics
Base Sequence
Binding Sites
/ genetics
Carotenoids
/ metabolism
Escherichia coli
/ genetics
Gene Expression Regulation, Bacterial
Glycolysis
/ genetics
Metabolic Engineering
/ methods
Metabolomics
/ methods
Models, Genetic
Promoter Regions, Genetic
/ genetics
Proteomics
/ methods
Repressor Proteins
/ genetics
Transcription Factors
/ genetics
Transcription, Genetic
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
13 08 2021
13 08 2021
Historique:
received:
23
07
2020
accepted:
21
07
2021
entrez:
14
8
2021
pubmed:
15
8
2021
medline:
31
8
2021
Statut:
epublish
Résumé
Synthetic metabolic pathways are a burden for engineered bacteria, but the underlying mechanisms often remain elusive. Here we show that the misregulated activity of the transcription factor Cra is responsible for the growth burden of glycerol overproducing E. coli. Glycerol production decreases the concentration of fructose-1,6-bisphoshate (FBP), which then activates Cra resulting in the downregulation of glycolytic enzymes and upregulation of gluconeogenesis enzymes. Because cells grow on glucose, the improper activation of gluconeogenesis and the concomitant inhibition of glycolysis likely impairs growth at higher induction of the glycerol pathway. We solve this misregulation by engineering a Cra-binding site in the promoter controlling the expression of the rate limiting enzyme of the glycerol pathway to maintain FBP levels sufficiently high. We show the broad applicability of this approach by engineering Cra-dependent regulation into a set of constitutive and inducible promoters, and use one of them to overproduce carotenoids in E. coli.
Identifiants
pubmed: 34389727
doi: 10.1038/s41467-021-25142-0
pii: 10.1038/s41467-021-25142-0
pmc: PMC8363753
doi:
Substances chimiques
Bacterial Proteins
0
Repressor Proteins
0
Transcription Factors
0
FruR protein, Bacteria
138186-82-0
Carotenoids
36-88-4
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4929Informations de copyright
© 2021. The Author(s).
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