Organic acid synthesis from C1 substrates
Presented herein are biocatalysts and methods for converting C1-containing materials to organic acids such as muconic acid or adipic acid.
1. An engineered cell, comprising an exogenously added gene encoding a 3-dehydroshikimate dehydratase (AroZ), a protocatechuic acid decarboxylase (AroY), a catechol 1,2-dioxygenase (CatA), a phospho-2-dehydro-3-deoxyheptonate aldolase (AroG), an anthranilate synthase (TrpE), a phosphoenolpyruvate synthase, or a transketolase; wherein the engineered cell is a methanotroph and is able to convert a C1 substrate to a muconic acid and the cell comprises genes encoding the 3-dehydroshikimate dehydratase (AroZ), the protocatechuic acid decarboxylase (AroY), and the catechol 1,2-dioxygenase (CatA).
2. The engineered cell of claim 1 , wherein the 3-dehydroshikimate dehydratase (AroZ), is from Klebsiella variicola.
3. The engineered cell of claim 1 , wherein the protocatechuic acid decarboxylase (AroY) is from Enterobacter cloacae.
4. The engineered cell of claim 1 , wherein the catechol 1,2-dioxygenase (CatA) is from Acinetobacter.
5. The engineered cell of claim 1 , wherein the 3-dehydroshikimate dehydratase (AroZ) is from Klebsiella variicola , the protocatechuic acid decarboxylase (AroY) is from Enterobacter cloacae , and the catechol 1,2-dioxygenase (CatA) is from Acinetobacter.
6. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a phosphoketolase.
7. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a CbbL, CbbS, CbbQ or CbbP polypeptide.
8. The engineered cell of claim 7 , wherein the cell further comprises exogenously added genes encoding CbbL, CbbS, CbbQ and CbbP polypeptides.
9. The engineered cell of claim 1 , wherein the cell is from an organism from the genus Methylococcus.
10. The engineered cell of claim 9 , wherein the cell is Methylococcus capsulatus.
11. The engineered cell of claim 1 , wherein the C1 substrate is methane, methanol, carbon dioxide, carbon monoxide, syngas, natural gas, or biogas.
12. A method for producing a muconic acid, comprising:
a) culturing the engineered cell of claim 1 with a C1 substrate; and
b) recovering the muconic acid from the culture.