Method for the incorporation of formaldehyde into biomass
Inventors
Bar-Even, Arren • HE, Hai • Marliére, Philippe
Assignees
Max-Planck-Gesellschaft zur Förderung der Wissenschaften eV • Scientist of Fortune SA
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Publication Number
US-12709764-B2
Publication Date
2026-08-18
Expiration Date
Abstract
The present disclosure relates to a method for the incorporation of formaldehyde into biomass comprising the following enzymatically catalyzed steps: (1) condensation of pyruvate with formaldehyde into 4-hydroxy-2-oxobutanoic acid (HOB); (2) amination of the thus produced 4-hydroxy-2-oxobutanoic acid (HOB) to produce homoserine; (3) conversion of thus produced homoserine to threonine; (4) conversion of the thus produced threonine into glycine and acetaldehyde or acetyl-CoA; (5) condensation of the thus produced glycine with formaldehyde to produce serine; and (6) conversion of the thus produced serine to produce pyruvate, wherein said pyruvate can then be used as a substrate in step (1). The disclosure also relates to enzymes for catalyzing the corresponding enzymatic reactions and recombinant microorganisms which express the enzymes for catalyzing the corresponding enzymatic reactions.
Core Innovation
A recombinant microorganism catalyzes, in vivo, a sequence of reactions for formaldehyde incorporation into carbon compounds assimilated into metabolism. The sequence includes condensation of pyruvate with a first formaldehyde into 4-hydroxy-2-oxobutanoic acid (HOB), conversion of HOB to homoserine, then to o-phosphohomoserine, and dephosphorylation to produce threonine.
The sequence continues by converting threonine into glycine using a threonine aldolase selected from aldolases classified in EC 4.1.2.5, EC 4.1.2.6, EC 4.1.2.48, or EC 4.1.2.49, or by a combination of a threonine dehydrogenase classified in EC 1.1.1.103 and a 2-amino-3-ketybutyrate CoA ligase classified in EC 2.3.1.29. The glycine is condensed with a second formaldehyde to produce serine by a threonine aldolase selected from the same listed EC 4.1.2.5, EC 4.1.2.6, EC 4.1.2.48, and EC 4.1.2.49, and serine is then deaminated to produce pyruvate by a serine deaminase classified in EC 4.3.1.17 or a threonine deaminase classified in EC 4.3.1.19.
The microorganism contains at least one heterologous nucleic acid molecule encoding the aldolase catalyzing step (1), and overexpresses the enzyme catalyzing step (6) for condensation of glycine with formaldehyde to form serine. The formaldehyde-assimilating reaction sequence also relates formaldehyde incorporation through a methanol-to-formaldehyde capability using methanol dehydrogenase (EC 1.1.1.244 or EC 1.1.2.7) and/or methanol oxidase (EC 1.1.3.13).
Claims Coverage
The partial content identifies one independent claim. This claim is supported by dependent claims that narrow the engineered microorganism, add additional enzyme overexpression steps, define additional formaldehyde supply, specify endogenous deficiencies, restrict the host organism, or broaden the claim to a cell-free extract implementation.
Recombinant microorganism with in vivo formaldehyde incorporation reaction sequence
A recombinant microorganism expresses enzymes for catalyzing, in vivo, a sequence of reactions for in vivo incorporation of formaldehyde into carbon compounds assimilated into metabolism, including condensation of pyruvate with a first formaldehyde into 4-hydroxy-2-oxobutanoic acid (HOB), conversion of HOB to homoserine and then to o-phosphohomoserine and threonine, conversion of threonine into glycine, condensation of glycine with a second formaldehyde to produce serine, and deamination of serine to produce pyruvate, wherein pyruvate can then be used as a substrate in step (1), and wherein the microorganism contains at least one heterologous nucleic acid molecule encoding the aldolase catalyzing step (1) and overexpresses the enzyme catalyzing step (6) for the condensation of glycine with formaldehyde to form serine.
Independent claim centers on an engineered recombinant microorganism that performs a defined in vivo formaldehyde-to-metabolism reaction sequence (pyruvate → HOB → homoserine → o-phosphohomoserine → threonine → glycine → serine → pyruvate), with required heterologous nucleic acid for the aldolase in step (1) and overexpression of the enzyme for condensation of glycine with formaldehyde to form serine.
Stated Advantages
Solving the problem of enabling in vivo incorporation of formaldehyde into carbon compounds assimilated into metabolism.
Documented Applications
In vivo incorporation of formaldehyde into carbon compounds assimilated into metabolism.
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