Fermentative production of alcohols
View Patent ↗The invention relates to the development of microorganisms capable of producing fermentation products via an engineered pathway in the microorganisms. The invention also relates to microorganisms with improved cell viability and methods to improve cell viability and cell productivity of a microorganism.
1. A recombinant yeast host cell comprising a modified polypeptide having yeast activator protein activity, wherein the modified polypeptide having yeast activator protein activity is Yap 1 and wherein the polypeptide comprises one or more amino acid substitutions corresponding to residues 411,584, and 636 of SEQ ID NO: 11 and the polypeptide comprises an insertion of one or more tripeptide repeat units selected from Ser-Thr-Asp and Ser-Asp-Gly.
2. The recombinant yeast host cell of claim 1 , wherein the polypeptide further comprises one or more amino acid substitutions corresponding to residues 80, 132, 168, 237, 254, 257, 297, 302, 367, 404, 444, 487, 498, 499, 548, or 617 of SEQ ID NO: 11.
3. The recombinant yeast host cell of claim 1 , wherein the polypeptide encoded by the nucleotide sequence in the recombinant yeast host cell comprises SEQ ID NO: 24.
4. The recombinant yeast host cell of claim 1 , wherein the recombinant yeast host cell further comprises a butanol biosynthetic pathway.
5. The recombinant yeast host cell of claim 4 , wherein the butanol biosynthetic pathway is an isobutanol biosynthetic pathway.
6. The recombinant yeast host cell of claim 4 , wherein the recombinant yeast host cell further comprises a deletion in an endogenous polynucleotide encoding a polypeptide having pyruvate decarboxylase activity.
7. The recombinant yeast host cell of claim 6 , wherein the polypeptide having pyruvate decarboxylase activity is selected from the group consisting of: PDC1, PDC5, PDC6, and combinations thereof.
8. The recombinant yeast host cell of claim 1 , wherein the recombinant yeast host cell is a member of a genus of yeast selected from the group consisting of: Saccharomyces, Schizosaccharomyces, Hansenula, Candida, Kluyveromyces, Yarrowia, Issatchenkia , and Pichia.
9. A composition comprising the recombinant yeast host cell of claim 1 .
10. The recombinant yeast host cell of claim 1 , wherein the one or more amino acid substitutions are N411S, D584G, or I636V.
11. The recombinant yeast host cell of claim 5 , wherein the isobutanol biosynthetic pathway comprises the following substrate to product conversions:
a) pyruvate to acetolactate;
b) acetolactate to 2,3-dihydroxyisovalerate;
c) 2,3-dihydroxyisovalerate to α-ketoisovalerate;
d) α-ketoisovalerate to isobutyraldehyde; and
e) isobutyraldehyde to isobutanol.
12. A process for producing an alcohol comprising
(a) providing the recombinant host cell of claim 1 , wherein the host cell produces an alcohol;
(b) contacting the recombinant host cell with one or more carbon substrates under conditions wherein the alcohol is produced; and
(c) recovering the alcohol.
13. The process of claim 12 , wherein the alcohol is selected from methanol, ethanol, propanol, butanol, pentanol, and hexanol.