Recombinant microorganisms having a methanol elongation cycle (MEC)
View Patent ↗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.
1. A recombinant prokaryote or yeast microorganism comprising a metabolic pathway for the synthesis of acetyl phosphate from methanol, methane or formaldehyde, wherein the microorganism is engineered to express the following enzymes: (a) a phosphoketolase (F/Xpk); (b) a transaldolase (Tal); (c) a transketolase (Tkt); (d) a ribose-5-phosphate isomerase (Rpi); (e) a ribulose-5-phosphate epimerase (Rpe); (f) a methanol dehydrogenase (Mdh); (g) a hexulose-6-phosphate synthase (Hps); (h) a hexulose-6-phosphate isomerase (Phi); (i) a dihydroxyacetone synthase (Das); and (j) a fructose-6-phosphate aldolase (Fsa), wherein at least one enzyme is heterologous to the microorganism, and wherein said microorganism has an acetyl-phosphate yield better than a wild-type or parental organism lacking Fpk and/or Xpk.
2. The recombinant microorganism of claim 1 , wherein the microorganism is derived from an E. coli microorganism.
3. The recombinant microorganism of claim 1 , wherein the microorganism is further engineered to express a phosphotransacetylase.
4. The recombinant microorganism of claim 1 , wherein the phosphoketolase comprises a sequence that is at least 49% identical to SEQ ID NO:2 and has phosphoketolase activity.
5. The recombinant microorganism of claim 1 , wherein the microorganism is E. coli comprising a genotype mdh, hps, phi and f/xpk.
6. The recombinant microorganism of claim 1 , wherein the pathway produces acetyl phosphate through carbon rearrangement of erythrose-4-phosphate (E4P) and/or glyceraldehyde-3-phosphate (G3P) and metabolism of a carbon source selected from methane, methanol, or formaldehyde.
7. The recombinant microorganism of claim 6 , wherein the microorganism is derived from an E. coli microorganism.
8. The recombinant microorganism of claim 7 , wherein the E. coli is engineered to express an Fpk, Xpk or a bifunctional F/Xpk phosphoketolase.
9. The recombinant microorganism of claim 8 , wherein the microorganism is further engineered to express a phosphotransacetylase.
10. The recombinant microorganism of claim 8 , wherein the phosphoketolase comprises a sequence that is at least 49% identical to SEQ ID NO:2 and has phosphoketolase activity.