Microorganisms and methods for enhancing the availability of reducing equivalents in the presence of methanol, and for producing succinate related thereto
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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Abstract
Provided herein is a non-naturally occurring microbial organism having a methanol metabolic pathway that can enhance the availability of reducing equivalents in the presence of methanol. Such reducing equivalents can be used to increase the product yield of organic compounds produced by the microbial organism, such as succinate. Also provided herein are methods for using such an organism to produce succinate.
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Citations
20 Claims
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1. A non-naturally occurring microbial organism comprising:
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(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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A non-naturally occurring microbial organism comprising:
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(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 consisting essentially of; (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. - View Dependent Claims (20)
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Specification