Journal article
Engineering a Synthetic Escherichia coli Coculture for Compartmentalized de novo Biosynthesis of Isobutyl Butyrate from Mixed Sugars
ACS synthetic biology, Vol.13(1), pp.259-268
01/19/2024
DOI: 10.1021/acssynbio.3c00493
PMCID: PMC10804408
PMID: 38091519
Abstract
Short-chain esters are versatile chemicals that can be used as flavors, fragrances, solvents, and fuels. The
ester biosynthesis consists of diverging and converging pathway submodules, which is challenging to engineer to achieve optimal metabolic fluxes and selective product synthesis. Compartmentalizing the pathway submodules into specialist cells that facilitate pathway modularization and labor division is a promising solution. Here, we engineered a synthetic
coculture with the compartmentalized sugar utilization and ester biosynthesis pathways to produce isobutyl butyrate from a mixture of glucose and xylose. To compartmentalize the sugar-utilizing pathway submodules, we engineered a xylose-utilizing
specialist that selectively consumes xylose over glucose and bypasses carbon catabolite repression (CCR) while leveraging the native CCR machinery to activate a glucose-utilizing
specialist. We found that the compartmentalization of sugar catabolism enabled simultaneous co-utilization of glucose and xylose by a coculture of the two
specialists, improving the stability of the coculture population. Next, we modularized the isobutyl butyrate pathway into the isobutanol, butyl-CoA, and ester condensation submodules, where we distributed the isobutanol submodule to the glucose-utilizing specialist and the other submodules to the xylose-utilizing specialist. Upon compartmentalization of the isobutyl butyrate pathway submodules into these sugar-utilizing specialist cells, a robust synthetic coculture was engineered to selectively produce isobutyl butyrate, reduce the biosynthesis of unwanted ester byproducts, and improve the production titer as compared to the monoculture.
Details
- Title: Subtitle
- Engineering a Synthetic Escherichia coli Coculture for Compartmentalized de novo Biosynthesis of Isobutyl Butyrate from Mixed Sugars
- Creators
- Hyeongmin Seo - University of Tennessee at KnoxvilleGillian Castro - University of Tennessee at KnoxvilleCong T Trinh - University of Tennessee at Knoxville
- Resource Type
- Journal article
- Publication Details
- ACS synthetic biology, Vol.13(1), pp.259-268
- DOI
- 10.1021/acssynbio.3c00493
- PMID
- 38091519
- PMCID
- PMC10804408
- ISSN
- 2161-5063
- eISSN
- 2161-5063
- Language
- English
- Date published
- 01/19/2024
- Academic Unit
- Chemical and Biochemical Engineering
- Record Identifier
- 9984696711802771
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