Competitive growth and/or production advantage for butanologen microorganism
View Patent ↗Provided herein are recombinant yeast host cells and methods for their use for production of fermentation products. Host cells provided herein comprise a pyruvate-utilizing pathway and a competitive growth advantage over other microorganisms in solution.
1. A method for production of isobutanol in a fermentation process comprising:
providing a fermentation mix comprising a recombinant yeast production microorganism which comprises an engineered isobutanol biosynthetic pathway, a heterologous polynucleotide encoding a polypeptide having acetolactate synthase activity, wherein said polypeptide has the amino acid sequence of SEQ ID NO: 9 and confers resistant to sulfonylureas, and a heterologous polynucleotide encoding a polypeptide having 3-phosphoshikimate 1-carboxylvinyltransferase activity; and
contacting the fermentation mix with at least one sulfonylurea which preferentially inhibits at least one contaminant yeast microorganism, wherein a fermentation product of the contaminant yeast microorganism is ethanol;
wherein production competitiveness of the recombinant yeast production microorganism is associated with a higher isobutanol-to-ethanol ratio as compared to a fermentation process without addition of one or more inhibitors, antibiotics, or combinations thereof.
2. The method of claim 1 , wherein the specific growth rate of the at least one contaminant yeast microorganism is reduced more than the specific growth rate of the recombinant yeast production microorganism.
3. The method of claim 1 , wherein production of the fermentation product of the at least one contaminant yeast microorganism is reduced more than the isobutanol production of the recombinant yeast production microorganism.
4. The method of claim 1 , wherein the contaminant yeast microorganism is Saccharomyces cerevisiae .
5. The method of claim 1 , wherein the sulfonylurea is an inhibitor of an ethanol biosynthesis pathway.
6. The method of claim 1 , wherein the sulfonylurea is an inhibitor of an amino acid biosynthesis pathway.
7. The method of claim 1 , wherein the sulfonylurea is selected from a group consisting of: nicosulfuron methyl, metsulfuron methyl, chlorimuron ethyl, sulfometuron methyl, chlorsulfuron, thifensulfuron methyl, and mixtures thereof.
8. The method of claim 1 , wherein the recombinant yeast production microorganism is selected from Schizosaccharomyces, Issatchenkia, Kluyveromyces, Yarrowia, Pichia, Candida, Hansenula, Aspergillus, Pachysolen, Rhodotorula, Zygosaccharomyces, Galactomyces, Torulaspora, Debayomyces, Williopsis, Dekkera, Kloeckera, Metschnikowia , and Saccharomyces .
9. The method of claim 1 , 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.
10. The method of claim 1 , wherein the recombinant yeast production microorganism further comprises one or more of the following modifications:
a deletion in one or more endogenous polynucleotides encoding a polypeptide having pyruvate decarboxylase activity;
a deletion, mutation, or substitution in an endogenous polynucleotide encoding a polypeptide having acetolactate reductase activity;
a deletion, mutation, or substitution in an endogenous polynucleotide encoding a polypeptide having aldehyde dehydrogenase activity;
a deletion in an endogenous polynucleotide encoding a polypeptide having hexokinase activity;
a deletion in an endogenous polynucleotide encoding a polypeptide having glycerol-3-phosphate dehydrogenase activity; or
a deletion in an endogenous gene encoding a polypeptide affecting Fe-S cluster biosynthesis, wherein the polypeptide is FRA2.