Abstract
The feasibility of 1,3-butanediol biosynthesis through the inverted cycle of fatty acid β-oxidation in Escherichia coli cells was investigated by the rational metabolic engineering approach. CoA-dependent aldehyde dehydrogenase MhpF and alcohol dehydrogenases FucO and YqhD were used as terminal enzymes catalyzing conversion of 3-hydroxybutyryl-CoA to 1,3-butanediol. Constitutive expression of the corresponding genes in E. coli strains, which are deficient in mixed acid fermentation pathways and expressing fàd regulon genes under control of P trc-ideal-4 promoter, did not lead to the synthesis of 1,3-butanediol during anaerobic glucose utilization. Additional inactivation of fadE and ydiO genes, encoding acyl-CoA dehydrogenases, also did not cause synthesis of the target product. Constitutive expression of aceEF-lpdA operon genes encoding enzymes of pyruvate dehydrogenase complex led to an increase in anaerobic synthesis of ethanol. Synthesis of 1,3-butanediol was observed with the overexpression of acetyl-CoA C-acetyltransferase AtoB. Constitutive expression of atoB gene in a strain with a basal expression of alcohol/aldehyde dehydrogenase leads to synthesis of 0.3 mM of 1,3-butanediol.
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Origina1,3-butanediol k]fatty acid β-oxidation k]Escherichia coli k]metabolic engineeringl Russian Text © A.Yu. Gulevich, A.Yu. Skorokhodova, A.A. Stasenko, R.S. Shakulov, V.G. Debabov, 2016, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2016, Vol. 52, No. 1, pp. 21–29.
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Gulevich, A.Y., Skorokhodova, A.Y., Stasenko, A.A. et al. Metabolic engineering of Escherichia coli for 1,3-butanediol biosynthesis through the inverted fatty acid β-oxidation cycle. Appl Biochem Microbiol 52, 15–22 (2016). https://doi.org/10.1134/S0003683816010051
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DOI: https://doi.org/10.1134/S0003683816010051