Metabolic engineering strategies to produce medium-chain oleochemicals via acyl-ACP:CoA transacylase activity

Yan, Qiang; Cordell, William T.; Jindra, Michael A.; Courtney, Dylan K.; Kuckuk, Madeline K.; Chen, Xuanqi; Pfleger, Brian F.

doi:10.1038/s41467-022-29218-3

Metabolic engineering strategies to produce medium-chain oleochemicals via acyl-ACP:CoA transacylase activity

Journal Article · Fri Mar 25 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-29218-3· OSTI ID:1857884

Yan, Qiang; Cordell, William T.; Jindra, Michael A.; Courtney, Dylan K.; Kuckuk, Madeline K.; Chen, Xuanqi;

Abstract

Microbial lipid metabolism is an attractive route for producing oleochemicals. The predominant strategy centers on heterologous thioesterases to synthesize desired chain-length fatty acids. To convert acids to oleochemicals (e.g., fatty alcohols, ketones), the narrowed fatty acid pool needs to be reactivated as coenzyme A thioesters at cost of one ATP per reactivation - an expense that could be saved if the acyl-chain was directly transferred from ACP- to CoA-thioester. Here, we demonstrate such an alternative acyl-transferase strategy by heterologous expression of PhaG, an enzyme first identified in Pseudomonads , that transfers 3-hydroxy acyl-chains between acyl-carrier protein and coenzyme A thioester forms for creating polyhydroxyalkanoate monomers. We use it to create a pool of acyl-CoA’s that can be redirected to oleochemical products. Through bioprospecting, mutagenesis, and metabolic engineering, we develop three strains of Escherichia coli capable of producing over 1 g/L of medium-chain free fatty acids, fatty alcohols, and methyl ketones.

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Research Organization:: Univ. of Illinois at Urbana-Champaign, IL (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018420

OSTI ID:: 1857884

Alternate ID(s):: OSTI ID: 1902706

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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Metabolic engineering strategies to produce medium-chain oleochemicals via acyl-ACP:CoA transacylase activity

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