Polypeptide and yeast cell compositions and methods of using the same

Patent metadata

Field	Value
Publication number	US-11725213-B2
Application number	US-202117341224-A
Country	US
Kind code	B2
Filing date	Jun 7, 2021
Priority date	Apr 6, 2018
Publication date	Aug 15, 2023
Grant date	Aug 15, 2023

Abstract

Polypeptides comprising maltose/maltotriose transporters are provided. Additionally, polynucleotides, DNA constructs, and vectors encoding a maltose/maltotriose transporter, or yeast cells harboring such polynucleotides are provided. The yeast cell may be a Saccharomyces eubayanus cell modified to increase the expression or transport activity of a maltose/maltotriose transporter at the plasma membrane of the cell. Further, methods are provided for making a fermentation product by culturing any one of the yeast cells described herein with a fermentable substrate. Finally, methods are provided to select for and isolate maltotriose-utilizing strains of Saccharomyces eubayanus.

First claim

We claim:

1. A DNA construct comprising a heterologous promoter operably linked to a polynucleotide encoding an alpha-glucoside transporter (AGT) polypeptide, wherein the AGT polypeptide has at least 95% sequence identity to SEQ ID NO: 7.

2. The DNA construct of claim 1 , wherein the promoter is a yeast promoter.

3. A yeast cell comprising the DNA construct of claim 1 .

4. A vector comprising the DNA construct of claim 1 .

5. The vector of claim 4 , wherein the vector comprises a plasmid.

6. A yeast cell comprising the vector of claim 4 .

7. A yeast cell that expresses an alpha-glucoside transporter (AGT) polypeptide having at least 95% sequence identity to SEQ ID NO: 7, wherein the yeast cell has increased maltose/maltotriose transport activity as compared to a control yeast cell.

8. The yeast cell of claim 7 , wherein the increased maltose/maltotriose transport activity is at the plasma membrane of the yeast cell.

9. The yeast cell of claim 7 , wherein the yeast cell comprises a DNA construct comprising a heterologous promoter operably linked to a polynucleotide encoding the AGT polypeptide.

10. The yeast cell of claim 7 , wherein the yeast cell comprises a vector comprising a DNA construct comprising a heterologous promoter operably linked to a polynucleotide encoding the AGT polypeptide.

11. The yeast cell of claim 7 , wherein the yeast cell is from the genus Saccharomyces.

12. The yeast cell of claim 11 , wherein the yeast cell is selected from the species consisting of Saccharomyces eubayanus, Saccharomyces cerevisiae, Saccharomyces paradoxus, Saccharomyces mikatae, Saccharomyces arboricola, Saccharomyces kudriavzevii, Saccharomyces jurei , and Saccharomyces uvarum.

13. The yeast cell of claim 7 , wherein the yeast cell is a Saccharomyces eubayanus cell.

14. A method for making a fermentation product comprising culturing the yeast cells of claim 7 with a fermentable substrate to produce the fermentation product.

15. The method of claim 14 , wherein the fermentable substrate comprises maltose or maltotriose.

16. The method of claim 14 , wherein the fermentable substrate comprises wort or malt extract.

17. The method of claim 14 , wherein the fermentation product is selected from the group consisting of a beer product, a wine product, an alcoholic beverage, a biochemical, and a biofuel.

Assignees

• Wisconsin Alumni Res Found

Inventors

• Hittinger Chris Todd
• Baker Emilyclare Patricia

Classifications

• C12N15/81 Primary

  for yeasts · CPC title

• C12N1/16

  Yeasts; Culture media therefor · CPC title

• C12P7/00

  Preparation of oxygen-containing organic compounds · CPC title

• C07K14/395

  from Saccharomyces · CPC title

• C12C11/00 Primary

  Fermentation processes for beer (preparation of wine C12G1/00) · CPC title