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Banerjee, Deepro; Jindra, Michael A.; Linot, Alec J.; Pfleger, Brian F.; Maranas, Costas D. (, Current Research in Biotechnology)
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Yan, Qiang; Cordell, William T.; Jindra, Michael A.; Courtney, Dylan K.; Kuckuk, Madeline K.; Chen, Xuanqi; Pfleger, Brian F. (, Nature Communications)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 inPseudomonads, 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 ofEscherichia colicapable of producing over 1 g/L of medium-chain free fatty acids, fatty alcohols, and methyl ketones.more » « less
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Hernández Lozada, NéstorJ.; Simmons, Trevor R.; Xu, Ke; Jindra, Michael A.; Pfleger, Brian F. (, Metabolic Engineering)
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Yan, Qiang; Simmons, Trevor R.; Cordell, William T.; Hernández Lozada, Néstor J.; Breckner, Christian J.; Chen, Xuanqi; Jindra, Michael A.; Pfleger, Brian F. (, Metabolic Engineering)
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