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Genome-wide screening identifies promiscuous phosphatases impairing terpenoid biosynthesis in Escherichia coli.
- Source :
-
Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2018 Nov; Vol. 102 (22), pp. 9771-9780. Date of Electronic Publication: 2018 Sep 18. - Publication Year :
- 2018
-
Abstract
- Terpenoids are a large family of natural compounds that are important for both biotechnological applications and basic microorganism physiology. Inspired by the current literature, we hypothesized that recently deciphered phosphatase promiscuity may be an unexplored factor that negatively affects terpenoid biosynthesis by redirecting carbon flux away from the pathway via unrecognized catalytic activities on the phosphorylated intermediates. We used lycopene as a proof-of-concept to test this hypothesis. Based on an extensive bioinformatics analysis, we selected 56 phosphatase-encoding genes in Escherichia coli and constructed a knockdown library for these genes in a lycopene overproducer via CRISPR interference (CRISPRi). We screened this phosphatase knockdown library and observed enrichment (28 of 56) for genes that impair lycopene biosynthesis. Further scaled-up cultivation, combinatorial knockdown, and knockout assays in strains that overproduce lycopene or another terpenoid (β-carotene) confirmed the proposed relationship between promiscuous phosphatases and impaired terpenoid biosynthesis. This study hence suggests the necessity of reconsidering the interactions of promiscuous phosphatases with ubiquitous phosphorylated components of metabolic networks with respect to engineering metabolism.
- Subjects :
- Biosynthetic Pathways
Escherichia coli chemistry
Escherichia coli genetics
Escherichia coli metabolism
Escherichia coli Proteins genetics
Metabolic Networks and Pathways
Phosphoric Monoester Hydrolases genetics
Terpenes chemistry
Escherichia coli enzymology
Escherichia coli Proteins metabolism
Genome, Bacterial
Phosphoric Monoester Hydrolases metabolism
Terpenes metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1432-0614
- Volume :
- 102
- Issue :
- 22
- Database :
- MEDLINE
- Journal :
- Applied microbiology and biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 30229323
- Full Text :
- https://doi.org/10.1007/s00253-018-9330-9