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Microorganisms and methods for enhancing the availability of reducing equivalents in the presence of methanol, and for producing succinate related thereto

  • US 10,640,795 B2
  • Filed: 02/23/2018
  • Issued: 05/05/2020
  • Est. Priority Date: 10/22/2012
  • Status: Active Grant
First Claim
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1. A non-naturally occurring microbial organism comprising:

  • (a) a methanol metabolic pathway, wherein said non-naturally occurring microbial organism comprises at least one exogenous nucleic acid encoding a methanol metabolic pathway enzyme expressed in a sufficient amount to enhance the availability of reducing equivalents in the presence of methanol or metabolize methanol as a carbon source for biosynthesis of fumarate, wherein said methanol metabolic pathway comprises;

    (i) a methanol methyltransferase and a methylenetetrahydrofolate reductase;

    (ii) a methanol dehydrogenase;

    or(iii) a methanol dehydrogenase and a formaldehyde activating enzyme; and

    (b) a fumarate pathway comprising exogenous nucleic acids wherein at least one of said exogenous nucleic acids is a heterologous nucleic acid encoding a fumarate pathway enzyme expressed in a sufficient amount to produce fumarate, said fumarate pathway comprising;

    (i) a phosphoenolpyruvate (PEP) carboxylase or a PEP carboxykinase, a malate dehydrogenase, and a fumarase;

    (ii) a pyruvate carboxylase, a malate dehydrogenase, and a fumarase;

    or(iii) a malic enzyme, and a fumarase.

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