Yeast alleles involved in maximal alcohol accumulation capacity and tolerance to high alcohol levels

Patent metadata

Field	Value
Publication number	US-10000759-B2
Application number	US-201414784893-A
Country	US
Kind code	B2
Filing date	Apr 15, 2014
Priority date	Apr 15, 2013
Publication date	Jun 19, 2018
Grant date	Jun 19, 2018

Abstract

The disclosure relates to a specific yeast allele of KIN3 that is involved in maximal alcohol accumulation and/or in tolerance to high alcohol levels. Preferably, the alcohol is ethanol. In a preferred embodiment, this specific allele is combined with specific alleles of ADE1 and/or VPS70. More specifically, the disclosure relates to the use of these alleles for the construction and/or selection of high alcohol tolerant yeasts, by stacking of positive alleles, or the selection and construction of low alcohol producing yeasts by stacking of negative alleles.

First claim

The invention claimed is:

1. A Saccharomyces cerevisiae yeast strain with higher alcohol accumulation produced by a method comprising:
selecting the strain for a protein kinase 3 (KIN3) allele that allows for a higher alcohol accumulation and that comprises at least a thymidine at position 553 and a thymidine at position 1648, wherein the positions refer to SEQ ID NO: 7;
selecting the strain for an adenine requiring 1 (ADE1) allele that comprises at least a thymidine at position 254, wherein the position refers to SEQ ID NO: 8;
and introducing into the strain nucleic acid molecule encoding the amino acid sequence of SEQ 10 NO: 4;
wherein the yeast strain produces increased alcohol concentration compared with alcohol concentration produced under identical conditions by a yeast that is genetically identical, apart from the selected KIN3 and ADE1 alleles and the introduced nucleic acid molecule.

2. The yeast strain of claim 1 , wherein the KIN3 allele consists of SEQ ID NO.1.

3. The yeast strain of claim 1 , further comprising at least one other alcohol tolerance modulating allele selected from the group consisting of vascuolar protein sorting 70 (VPS70), anti-prion dnaJ (APJ1) and sick without securin 2 (SWS2).

4. The yeast strain of claim 2 , further comprising at least one other alcohol tolerance modulating allele selected from the group consisting of vacuolar protein sorting 70 (VPS70), anti-prion dnaJ (APJ1) and sick without securin 2 (SWS2).

5. The yeast strain of claim 4 , further comprising an inactive APJ1 allelee.

6. The yeast strain of claim 4 , further comprising an SWS2 allele that overexpresses SWS2 protein.

7. The yeast strain of claim 3 , further comprising at least one allele selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, and SEQ ID NO:6.

8. The yeast strain of claim 7 , further comprising the combination of SEQ ID NO:3 with a nucleic acid molecule encoding SEQ ID NO:4.

9. A process for producing ethanol, the process comprising: culturing the saccharomyces cerevisiae yeast strain of claim 1 to ferment sugars to ethanol.

10. The yeast strain of claim 1 , wherein the ADE1 allele consists of SEQ ID NO:2.

11. A Saccharomyces cerevisiae yeast strain comprising:
a protein kinase 3 (KIN3) allele having a thymidine at position 553 and a thymidine at position 1648, wherein the positions refer to SEQ ID NO: 7;
an adenine requiring 1 (ADE1) allele that comprises a thymidine at position 254; wherein the position refers to in SEQ ID NO: 8;
and a nucleic acid molecule encoding the amino acid sequence of SEQ ID NO: 4.

Assignees

• Vib Vzw
• Univ Leuven Kath
• Univ Leuven Kath

Inventors

• Thevelein Johan
• Goovaerts Annelies
• Dumortier Françoise
• Foulquie-Moreno Maria Remedios
• Swinnen Steve
• Martins Pais Thiago

Classifications

• C12P7/06

  Ethanol, i.e. non-beverage · CPC title

• C07K14/395 Primary

  from Saccharomyces · CPC title

• C12N15/81 Primary

  for yeasts · CPC title

• Y02E50/17

  Cross-Sectional Technologies · mapped topic

• Y02E50/10

  Biofuels, e.g. bio-diesel · CPC title