Recombinant microorganisms having a methanol elongation cycle (mec)
AssigneeThe Regents of the University of California
InventorsJames C. Liao, Igor Bogorad
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Abstract
Provided are microorganisms that catalyze the synthesis of chemicals and biochemicals from a methanol, methane and/or formaldehyde. Also provided are methods of generating such organisms and methods of synthesizing chemicals and biochemicals using such organisms.
Claims47
A recombinant microorganism comprising a metabolic pathway for the synthesis of acetyl phosphate from methanol, methane or formaldehyde using a pathway comprising an enzyme having fructose-6-phosphoketolase (Fpk) activity and/or xylulose-5-phosphoketolase (Xpk) activity with an acetyl-phosphate yield better than a wild-type or parental organism lacking Fpk and/or Xpk.
A recombinant microorganism comprising a pathway that produces acetyl-phosphate through carbon rearrangement of E4P and/or G3P and metabolism of a carbon source selected from methane, methanol, or formaldehyde.
A recombinant microorganism expressing enzymes that catalyze the conversion described in (i)-(xi), wherein at least one enzyme or the regulation of at least one enzyme that performs a conversion described in (i)-(xi) is heterologous to the microorganism:
- (i) the production of acetyl-phosphate and erythrose-4-phosphate (E4P) from fructose-6-phosphate and/or the production of acetyl-phosphate and glyceraldehyde 3-phosphate (G3P) from xylulose 5-phosphate;
- (ii) the reversible conversion of fructose-6-phosphate and E4P to sedoheptulose 7-phosphate (S7P) and (G3P);
- (iii) the reversible conversion of S7P and G3P to ribose-5-phosphate and xylulose-5-phosphate;
- (iv) the reversible conversion of ribose-5-phosphate to ribulose-5-phosphate;
- (v) the reversible conversion of ribulose-5-phosphate to xylulose-5-phosphate;
- (vi) the reversible conversion of xylulose-5-phosphate and E4P to fructose-6-phosphate and glyceraldehyde-3-phosphate;
- (vii) the conversion of formaldehyde and ribulose-5-phosphate to D-arabino-3-Hexulose 6-phosphate;
- (viii) the reversible conversion of D-arabino-3-Hexulose 6-phosphate to fructose-6-phosphate;
- (ix) the conversion of formaldehyde and xylulose-5-phosphate to glyceraldehyde-3-phosphate and dihydroxyacetone;
- (x) the conversion of glyceraldehyde-3-phosphate and dihydroxyacetone to fructose-6-phosphate;
- and
- (xi) the conversion of methanol and a oxidized electron acceptor to formaldehyde and a reduced electron acceptor;
- wherein the microorganism produces acetyl-phosphate, or compounds derived from acetyl-phosphate using a carbon source selected from the group consisting of methanol, methane, and formaldehyde and any combination thereof.
A recombinant E. coli that produces acetyl-phosphate comprising a genotype mdh, hps, phi, and f/xpk.
A recombinant yeast that produces acetyl-phosphate comprising a genotype mdh, hps, phi, and f/xpk.
A recombinant Bacillus methanolicus that produces acetyl-phosphate comprising a genotype f/xpk.
A recombinant microorganism comprising a metabolic pathway for the synthesis of acetyl phosphate from methanol, methane or formaldehyde using a pathway comprising an enzyme having fructose-6-phosphoketolase (Fpk) activity and/or xylulose-5-phosphoketolase (Xpk) activity and wherein the microorganism produces a metabolite selected from the group consisting of citrate, isocitrate, alpha-ketoglutarate, glutamate and any combination thereof.
Dependencies are inferred from the claim text.