Microorganisms and methods for enhancing the availability of reducing equivalents in the presence of methanol, and for producing 1,4-butanediol related thereto

A method of producing an intermediate of a metabolic pathway that can be used in the formation of biomass, comprising culturing a non-naturally occurring microbial organism under conditions and for a sufficient period of time to produce the intermediate,

• wherein the non-naturally occurring microbial organism has a methanol metabolic pathway and comprises at least three exogenous nucleic acids each 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 formaldehyde, wherein said methanol metabolic pathway comprises:
• (i) a methanol methyltransferase, which catalyzes the conversion of methanol to methyltetrahydrofolate (methyl-THF),
• (ii) a methylenetetrahydrofolate reductase, which catalyzes the conversion of methyl-THF to methylene-THF, and
• (iii) a methylenetetrahydrofolate dehydrogenase, which catalyzes the conversion of methylene-THF to methenyl-THF;
• and
• wherein the non-naturally occurring microbial organism has a formaldehyde assimilation pathway and comprises at least two exogenous nucleic acids each encoding a formaldehyde assimilation pathway enzyme expressed in a sufficient amount to produce an intermediate of a metabolic pathway that can be used in the formation of biomass, wherein the formaldehyde assimilation pathway comprises:
• (i) a hexulose-6-phosphate synthase and a 6-phospho-3-hexuloisomerase;
• or
• (ii) a dihydroxyacetone synthase and a dihydroxyacetone kinase.