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Study of the Potential of the Reversal of the Fatty-Acid Beta-Oxidation Pathway for Stereoselective Biosynthesis of (S)-1,3-Butanediol from Glucose by Recombinant Escherichia coli Strains

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Abstract

The possible contribution of collateral enzymes to the formation of the key precursor metabolite, 3-hydroxybutyryl-CoA, has been evaluated in a recombinant Escherichia coli strain engineered for 1,3-butanediol biosynthesis from glucose via the inverted fatty-acid beta-oxidation pathway. Inactivation of the 3-hydroxyadipyl-CoA dehydrogenase gene, paaH, did not prevent 1,3-butanol biosynthesis during anaerobic glucose utilization by a strain with an intact, essential gene, fabG. This gene encodes 3-ketoacyl-ACP reductase, which can catalyze the conversion of acetoacetyl-CoA to (R)-3-hydroxybutyryl-CoA. The subsequent inactivation in the strain of the fadB gene, which encodes (S)-stereospecific 3-hydroxyacyl-CoA dehydrogenase of the fatty-acid beta oxidation led to the cessation of 1,3-butanediol synthesis. The respective diol was also not found among the products secreted by the strain possessing the intact fabG and paaH genes upon the individual deletion of the fadB gene. It was established that the collateral enzymes did not participate in the formation of 3-hydroxybutyryl-CoA in the studied strains, and the respective CoA derivative was synthesized solely by the (S)-specific enzyme of the fatty-acid beta-oxidation pathway. The results indicate that reversal of the fatty-acid beta oxidation pathway can ensure the enantioselective biosynthesis of the (S)-stereoisomer of 1,3-butanediol in engineered E. coli strains.

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Funding

The work was supported by a grant from the Russian Foundation for Basic Research (project no.18-29-08059).

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Authors and Affiliations

  1. Kurchatov Institute National Research Center, State Research Institute for Genetics and Selection of Industrial Microorganisms (Kurchatov Institute NRC—GOSNIIgenetika), 117545, Moscow, Russia

    A. Yu. Gulevich, A. Yu. Skorokhodova & V. G. Debabov

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  1. A. Yu. Gulevich
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  2. A. Yu. Skorokhodova
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  3. V. G. Debabov
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Correspondence to A. Yu. Gulevich.

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Translated by I. Gordon

Abbreviations: ACP—acyl carrier protein; HPLC—high performance liquid chromatography; IPTG—isopropyl-β-D-thiogalactoside; PCR—polymerase chain reaction.

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Gulevich, A.Y., Skorokhodova, A.Y. & Debabov, V.G. Study of the Potential of the Reversal of the Fatty-Acid Beta-Oxidation Pathway for Stereoselective Biosynthesis of (S)-1,3-Butanediol from Glucose by Recombinant Escherichia coli Strains. Appl Biochem Microbiol 56, 822–827 (2020). https://doi.org/10.1134/S0003683820080049

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