Compositions and methods for 3-hydroxypropionic acid production
The present application discloses genetically modified yeast cells comprising an active 3-HP fermentation pathway, and the use of these cells to produce 3-HP.
1. A genetically modified yeast cell comprising
one or more 3-HP pathway genes selected from the group consisting of:
an exogenous gene encoding an enzymatically active polypeptide that catalyzes the conversion of pyruvate to oxaloacetate (OAA);
an exogenous gene encoding an enzymatically active polypeptide that catalyzes the conversion of aspartate to β-alanine; and
an exogenous gene encoding an enzymatically active polypeptide that catalyzes the conversion of β-alanine to malonate semialdehyde and comprises at least 85% sequence identity to an amino acid sequence from SEQ ID NO: 20, 21, or 24.
2. The genetically modified yeast cell of claim 1 , wherein the yeast cell is selected from a Crabtree-negative yeast, an Issatchenkia yeast, a Candida yeast, a Kluyveromyces yeast, a Pichia yeast, a Schizosaccharomyces yeast, a Torulaspora yeast, a Zygosaccharomyces yeast, or a Saccharomyces yeast.
3. The genetically modified yeast cell of claim 2 , wherein the yeast cell is selected from the group consisting of Issatchenkia orientalis, Candida lambica , and Saccharomyces bulderi.
4. The genetically modified yeast cell of claim 1 , wherein the exogenous gene encoding the enzymatically active polypeptide that catalyzes the conversion of aspartate to β-alanine comprises an aspartate decarboxylase (ADC) gene that encodes a polypeptide with at least 50% sequence identity to an amino acid sequence from SEQ ID NO: 17, 18, 133, 135, 137, or 139.
5. The genetically modified yeast cell of claim 1 , wherein the genetically modified yeast cell further comprises an exogenous gene encoding an enzymatically active polypeptide that catalyzes the conversion of oxaloacetate (OAA) to aspartate.
6. The genetically modified yeast cell of claim 5 , wherein the exogenous gene encoding the enzymatically active polypeptide that catalyzes the conversion of oxaloacetate (OAA) to aspartate comprises an aspartate aminotransferase AAT gene that encodes a polypeptide with at least 50% sequence identity to an amino acid sequence from SEQ ID NO: 14, 15, or 16.
7. The genetically modified yeast cell of claim 1 , wherein the exogenous gene encoding the enzymatically active polypeptide that catalyzes the conversion of pyruvate to oxaloacetate (OAA) comprises an pyruvate carboxylase gene PYC gene that encodes a polypeptide with at least 50% sequence identity to an amino acid sequence from SEQ ID NO: 2, 3, 4, 5, 6, 7, or 8.
8. The genetically modified yeast cell of claim 1 , wherein the genetically modified yeast cell further comprises an exogenous gene encoding an enzymatically active polypeptide that catalyzes the conversion of phosphoenoylpyruvate (PEP) to oxaloacetate (OAA).
9. The genetically modified yeast cell of claim 8 , wherein the exogenous gene encoding the enzymatically active polypeptide that catalyzes the conversion of phosphoenoylpyruvate (PEP) to oxaloacetate (OAA) comprises an PEP carboxylase gene (PCK) that encodes a polypeptide comprising an amino acid sequence from SEQ ID NO: 35, 36, 37, 38, or 39.
10. The genetically modified yeast cell of claim 1 , wherein the genetically modified yeast cell further comprises a deletion or disruption of one or more native genes involved in ethanol fermentation.
11. The genetically modified yeast cell of claim 10 , wherein the one or more native genes involved in ethanol fermentation comprises pyruvate decarboxylase (PDC) or alcohol dehydrogenase (ADH).
12. The genetically modified yeast cell of claim 1 , wherein the genetically modified yeast cell further comprises a deletion or disruption of one or more native genes encoding glycerol 3-phosphate dehydrogenase (GPD), glycerol 3-phosphatase (GPP), glycerol kinase, dihydroxyacetone kinase, glycerol dehydrogenase, aldehyde dehydrogenase (ALD), or butanediol dehydrogenase.