Fermentation process using yeast cells having disrupted pathway from dihydroxyacetone phosphate to glycerol

Patent metadata

Field	Value
Publication number	US-10899544-B2
Application number	US-201815941274-A
Country	US
Kind code	B2
Filing date	Mar 30, 2018
Priority date	Mar 13, 2006
Publication date	Jan 26, 2021
Grant date	Jan 26, 2021

Abstract

Yeast cells are genetically modified to disrupt a native metabolic pathway from dihydroxyacetone to glycerol. In certain aspects, the yeast cell is of the genera Kluyveromyces, Candida or Issatchenkia . In other aspects, the yeast cell is capable of producing at least one organic acid, such as lactate. The yeast cells produce significantly less glycerol than the wild-type strains, and usually produce greater yields of desired fermentation products. Yeast cells of the invention often grow well when cultivated, despite their curtailed glycerol production.

First claim

The invention claimed is:

1. A fermentation process wherein a cell is cultivated in a fermentation medium under fermentation conditions and in the presence of a carbon source and added glycerol to produce an organic acid or ethanol, wherein the glycerol yield is less than 2% based on the weight of the carbon source that is consumed by the cell, wherein the cell is a yeast cell selected from the group consisting of Issatchenkia orientalis, Pichia galeiformis, Pichia sp. YB-4149 (NRRL designation), Candida ethanolica, P. deserticola, P. membranifaciens, P. fermentans, S. kluyveri, K. aestuaryii, K. nonfermentans, K. lactic, K. marxianus and K. dobzhanskii , in which the entire coding region of at least one native glycerol-3-phosphate dehydrogenase gene has been eliminated or the coding region, promoter or terminator region of at least one native glycerol-3-phosphate dehydrogenase gene has been modified by deletion or insertion so the native glycerol-3-phosphate dehydrogenase gene no longer produces an active glycerol-3-phosphate dehydrogenase enzyme.

2. The fermentation process of claim 1 wherein the organic acid or ethanol is produced in a yield of at least 40% based on the weight of the carbon source that is consumed by the cell.

3. The process of claim 2 , wherein the glycerol yield is less than 0.5% based on the weight of the carbon source that is consumed by the cell.

4. The process of claim 3 , wherein ethanol is produced in the fermentation process.

5. The fermentation process of claim 1 wherein an organic acid is produced in the fermentation process in a yield of at least 40% based on the weight of the carbon source that is consumed by the cell and the glycerol yield is less than 0.5% based on the weight of the carbon source that is consumed by the cell.

6. The fermentation process of claim 1 , wherein the cell is Issatchenkia orientalis.

7. The fermentation process of claim 1 , wherein the cell is Kluyveromyces marxianus.

8. The fermentation process of claim 5 , wherein the cell is Kluyveromyces marxianus or Issatchenkia orientalis.

9. A fermentation process wherein a yeast cell is cultivated in a fermentation medium under fermentation conditions and in the presence of a carbon source and added glycerol to produce an organic acid, wherein the glycerol yield is less than 2% based on the weight of the carbon source that is consumed by the cell, wherein the yeast cell is selected from a species of the group consisting of Issatchenkia orientalis, Pichia galeiformis, Pichia sp. YB-4149 (NRRL designation), Candida ethanolica, P. deserticola, P. membranifaciens, P. fermentans, S. kluyveri, K. aestuaryii, K. nonfermentans, K. lactic, K. marxianus and K. dobzhanskii , and the yeast cell has a deletion or disruption of a native metabolic pathway from dihydroxyacetone phosphate to glycerol, wherein the deletion or disruption of the native metabolic pathway includes (i) a deletion or disruption of at least one native glycerol-3-phophate dehydrogenase; (ii) a deletion or disruption of at least one native glycerol-3-phosphatase gene; (iii) a deletion or disruption of at least one native glycerol-3-phosphate dehydrogenase gene and of at least one native glycerol-3-phosphatase gene; (iv) a deletion or disruption of at least one native dihydroxyacetone phosphatase gene; or (v) a deletion or disruption of both a native dihydroxyacetone phosphatase gene and a native glycerol-3-phosphate dehydrogenase gene; wherein the yeast cell produces an organic acid.

Assignees

• Cargill Inc

Inventors

• Dundon Catherine Asleson
• Suominen Pirrko
• Aristidou Aristos
• Rush Brian J
• Koivuranta Kari
• Hause Benjamin Matthew
• Mcmullin Thomas William
• Roberg-Perez Kevin

Classifications

• C12N15/52 Primary

  Genes encoding for enzymes or proenzymes · CPC title

• C12P7/06

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

• C12P7/40

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title

• B65G17/12

  comprising a series of individual load-carriers fixed, or normally fixed, relative to traction element · CPC title

• Y02E50/10

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