Method for the incorporation of formaldehyde into biomass
AssigneesScientist of Fortune, S.A., Max-Planck-Gesellschaft zur Forderung der Wissenschaften e.V.
InventorsArren BAR-EVEN, Hai HE, Philippe MARLIÈRE
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Abstract
Described is 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).
Claims14
A method 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).
A recombinant microorganism expressing enzymes for catalyzing the following reactions:
- (1) condensation of pyruvate with formaldehyde into 4-hydroxy-2-oxobutanoic acid (HOB) by an aldolase classified in EC 4.1.2._;
- (2) amination of the thus produced 4-hydroxy-2-oxobutanoic acid (HOB) to produce homoserine by an aminotransferase enzyme classified in EC 2.6.1._ or by an amino acid dehydrogenase (EC 1.4.1._);
- (3) phosphorylation of thus produced homoserine to produce o-phosphohomoserine by a homoserine kinase (EC 2.7.1.39);
- (4) dephosphorylation of the thus produced o-phosphohomoserine to produce threonine by a threonine synthase (EC 4.2.3.1);
- (5) conversion of the thus produced threonine into glycine by a threonine aldolase (selected from the group consisting of EC 4.1.2.5, EC 4.1.2.6, EC 4.1.2.48 and EC 4.1.2.49) or by a combination of a threonine dehydrogenase (EC 1.1.1.103) and a 2-amino-3-ketybutyrate CoA ligase (EC 2.3.1.29);
- (6) condensation of the thus produced glycine with formaldehyde to produce serine by a threonine aldolase (selected from the group consisting of EC 4.1.2.5, EC 4.1.2.6, EC 4.1.2.48 and EC 4.1.2.49);
- and
- (7) deamination of the thus produced serine to produce pyruvate by a serine deaminase (EC 4.3.1.17 or a threonine deaminase (EC 4.3.1.19)), wherein said pyruvate can then be used as a substrate in step (1),
- wherein said microorganism contains at least one heterologous nucleic acid molecule encoding the aldolase catalyzing step (1) and
- overexpresses the enzyme catalyzing step (6), i.e. the condensation of glycine with formaldehyde to form serine.
Dependencies are inferred from the claim text.