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

Qiang Yan, William T. Cordell, Michael A. Jindra, Dylan K. Courtney, Madeline K. Kuckuk, Xuanqi Chen and Brian F. Pfleger ()
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Qiang Yan: University of Wisconsin-Madison
William T. Cordell: University of Wisconsin-Madison
Michael A. Jindra: University of Wisconsin-Madison
Dylan K. Courtney: University of Wisconsin-Madison
Madeline K. Kuckuk: University of Wisconsin-Madison
Xuanqi Chen: University of Wisconsin-Madison
Brian F. Pfleger: University of Wisconsin-Madison

Nature Communications, 2022, vol. 13, issue 1, 1-10

Abstract: 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.

Date: 2022
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DOI: 10.1038/s41467-022-29218-3

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