Engineering a Synthetic Coculture for Compartmentalized Biosynthesis of Isobutyl Butyrate from Mixed Sugars

Overview

Journal ACS Synth Biol

Publisher American Chemical Society

Specialties Biotechnology
Molecular Biology

Date 2023 Dec 13

PMID 38091519

Authors

Hyeongmin Seo

Gillian Castro

Cong T Trinh

Affiliations

Soon will be listed here.

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.

Citing Articles

Strategies and tools to construct stable and efficient artificial coculture systems as biosynthetic platforms for biomass conversion.

Song X, Ju Y, Chen L, Zhang W Biotechnol Biofuels Bioprod. 2024; 17(1):148.

PMID: 39702246 PMC: 11660635. DOI: 10.1186/s13068-024-02594-2.

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