Systems and methods for biotransformation of carbon dioxide into higher carbon compounds
AssigneeUniversity of Washington through its Center for Commercialization
InventorsJustin Siegel, David Baker, Amanda Lee Smith, Mary E. Lidstrom, Catherine Louw
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Abstract
Systems, compounds and methods for the conversion of C1 carbon compounds to higher carbon compounds useful for the generation of commodity compounds.
Claims19
A method for converting formaldehyde to dihydroxyacetone, comprising contacting formaldehyde with a thiamine diphosphate cofactor and a polypeptide comprising a motif capable of forming hydrogen bonds with the thiamine diphosphate cofactor,
- wherein the polypeptide comprises a sequence with at least 80% identity to the benzaldehyde lyase sequence set forth in SEQ ID NO:2;
- wherein the motif comprises a histidine residue in a first position corresponding to amino acid residue position 29 of SEQ ID NO:2, an asparagine or a glutamine residue in a second position corresponding to amino acid residue position 113 of SEQ ID NO:2, and a glutamic acid or an aspartic acid residue in a third position corresponding to amino acid residue position 50 of SEQ ID NO:2;
- wherein upon association with the thiamine diphosphate cofactor, the cofactor pyrimidine N1 is hydrogen bonded to the acid side chain of the glutamic acid residue or the acid side chain of the aspartic acid residue of the motif;
- and
- wherein the histidine N1 and the amide side chain of the asparagine residue or the amide side chain of the glutamine residue are capable of making a water-mediated hydrogen bond with the formaldehyde substrate and/or dihydroxyacetone product.
An engineered polypeptide to convert formaldehyde to dihydroxyacetone, comprising a motif capable of forming hydrogen bonds with a thiamine diphosphate cofactor,
- wherein the polypeptide comprises a sequence with at least 80% identity to the benzaldehyde lyase sequence set forth in SEQ ID NO:2;
- wherein the motif comprises a histidine residue in a first position corresponding to amino acid residue position 29 of SEQ ID NO:2, an asparagine or a glutamine residue in a second position corresponding to amino acid residue position 113 of SEQ ID NO:2, and a glutamic acid or an aspartic acid residue in a third position corresponding to amino acid residue position 50 of SEQ ID NO:2, and the motif further comprises at least one amino acid substitution corresponding to the substitutions selected from the group consisting of A394G, A480W, G419N, and A28I, with reference to the amino acid sequence set forth in SEQ ID NO:2;
- wherein upon association with the thiamine diphosphate cofactor, the cofactor pyrimidine N1 is hydrogen bonded to the acid side chain of the glutamic acid residue or the acid side chain of the aspartic acid residue of the motif;
- and
- wherein the histidine N1 and the amide side chain of the asparagine residue or the amide side chain of the glutamine residue are capable of making a water-mediated hydrogen bond with the formaldehyde substrate and/or dihydroxyacetone product.
Dependencies are inferred from the claim text.