Biocatalytic production of para-hydroxybenzoic acid from methanol and methane
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorNils AVERESCH
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Abstract
A method of producing para-hydroxybenzoic acid (pHBA) or a derivative thereof includes culturing the recombinant microorganism in a fermentation broth, wherein said recombinant microorganism comprising a genetically engineered pathway expressing at least one nucleic acid sequence encoding a polypeptide selected from: an exogenous chorismate pyruvate lyase of EC 5.4.4.2 or EC 4.1.3.40; an exogenous 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase of EC 4.1.2.15, or EC 2.5.1.54; an exogenous shikimate kinase of EC 2.7.1.71; or an exogenous 3-dehydroquinate dehydratase (DHQ) of EC 4.2.1.10; adding a carbon source to the fermentation broth; and isolating the pHBA from the fermentation broth.
Claims30
A method of producing para-hydroxybenzoic acid (pHBA) or a derivative thereof, the method comprising:
- culturing a recombinant microorganism in a fermentation broth;
- adding a carbon source to the fermentation broth;
- and
- isolating the pHBA from the fermentation broth;
- wherein the recombinant microorganism comprises a genetically engineered pathway comprising a nucleic acid sequence encoding an exogenous 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase of EC 4.1.2.15, or EC 2.5.1.54;
- and
- wherein the exogenous DAHP synthase of EC 4.1.2.15, or EC 2.5.1.54 comprises a feedback-inhibition resistant mutation in the wild-type amino acid sequence of the DAHP synthase.
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The method of claim 48, wherein
- the chorismate pyruvate lyase is P. rustigianii UbiC, C. sakazakii UbiC, or comprises the amino acid sequence of SEQ ID NO:
- 4 or 5;
- the DAHP synthase is E. coli AroG, or comprises the amino acid sequence of SEQ ID NO:
- 1;
- the shikimate kinase is E. coli AroL, or comprises the amino acid sequence of SEQ ID NO:
- 3;
- or
- the DHQ is E. coli AroD, or comprises the amino acid sequence of SEQ ID NO:
- 2.
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A recombinant microorganism for producing para-hydroxybenzoic acid (pHBA) or a derivative thereof, the recombinant microorganism comprising a genetically engineered pathway comprising a nucleic acid sequence encoding an exogenous 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase of EC 4.1.2.15, or EC 2.5.1.54,
- wherein the exogenous DAHP synthase of EC 4.1.2.15, or EC 2.5.1.54 comprises a feedback-inhibition resistant mutation in the wild-type amino acid sequence of the DAHP synthase.
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A vector comprising a nucleic acid sequence set forth in SEQ ID NOs:
- 6, 45, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, or a combination of any two or more thereof.
Dependencies are inferred from the claim text.