Cell-associated heterologous food and/or feed enzymes

Patent metadata

Field	Value
Publication number	US-12344847-B2
Application number	US-201816493245-A
Country	US
Kind code	B2
Filing date	Mar 13, 2018
Priority date	Mar 13, 2017
Publication date	Jul 1, 2025
Grant date	Jul 1, 2025

Abstract

The present disclosure concerns recombinant yeast host cells expressing cell-associated heterologous food and/or feed enzymes which are expressed during the propagation phase of the recombinant yeast hosts cells. The recombinant yeast host cells can be used in a subsequent production process to make food and/or feed products, for example, baked products.

First claim

What is claimed is:

1. A process for making a food or a feed product, said process comprising including a recombinant yeast host cell or an additive comprising the recombinant yeast host cell in the food or the feed product, wherein the recombinant yeast host cell has a heterologous nucleic acid molecule encoding a cell-associated heterologous maltogenic alpha-amylase, wherein the maltogenic alpha-amylase has at least 80% identity to the amino acid sequence of SEQ ID NO: 1, 51, 65 or 108, wherein the heterologous nucleic acid molecule is operatively associated with a promoter allowing expression of the heterologous nucleic acid molecule during propagation under aerobic conditions.

2. The process of claim 1 , wherein the heterologous nucleic acid molecule allows intracellular expression of the heterologous maltogenic alpha-amylase.

3. The process of claim 1 , wherein the heterologous nucleic acid molecule allows expression of a membrane-associated heterologous maltogenic alpha-amylase.

4. The process of claim 1 , wherein the heterologous nucleic acid molecule allows expression of a tethered heterologous maltogenic alpha-amylase.

5. The process of claim 4 , wherein the tethered heterologous maltogenic alpha-amylase is a chimeric protein of formula (I) or (II):
(NH 2 )FFE-L-TT(COOH)  (I)

(NH 2 )TT-L-FFE(COOH)  (II)

wherein FFE is the maltogenic alpha-amylase;
L is present or absent and is an amino acid linker;
TT is an amino acid tethering moiety for associating the maltogenic alpha-amylase to a cell wall of the recombinant yeast host cell;
(NH2) indicates the amino terminus of the chimeric protein;
(COOH) indicates the carboxyl terminus of the chimeric protein; and
“-” is an amide linkage.

6. The process of claim 1 , wherein the promoter is a heterologous promoter.

7. The process of claim 6 , wherein the heterologous promoter comprises the promoter from the tdh1 gene, the hor7 gene, the hsp150 gene, the hxt7 gene, the gpm1 gene, the pgk1 gene and/or the stl1 gene.

8. The process of claim 1 , wherein the recombinant yeast host cell is from the genus Saccharomyces sp.

9. The process of claim 1 , wherein the recombinant yeast host cell is from the species Saccharomyces cerevisiae.

10. The process of claim 1 , wherein the additive is a food additive, a dough conditioner, or a feed additive.

11. The process of claim 1 , further comprising fermenting the food or the feed product with the recombinant yeast host cell present, and/or baking the food or the feed product to provide a baked product.

12. The process of claim 11 , wherein the baked product is a bread.

13. The process of claim 3 , wherein the membrane-associated heterologous maltogenic alpha-amylase has a heterologous signal peptide.

14. The process of claim 4 , wherein the tethered heterologous maltogenic alpha-amylase has a heterologous signal peptide.

15. The process of claim 1 , wherein the promoter is a native promoter.

Assignees

• Lallemand Hungary Liquidity Man Llc
• Danstar Ferment Ag

Inventors

• Argyros Aaron
• Oeser Michelle
• Wiswall Erin
• Fisher Janet
• Van Eijk Johannes
• Kraus J Kevin
• Wenger Kevin
• Henningsen Brooks
• Skinner Ryan

Classifications

• C12Y302/01133

  Glucan 1,4-alpha-maltohydrolase (3.2.1.133), i.e. maltogenic alpha-amylase · CPC title

• C12N9/2428

  Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase · CPC title

• C12N9/242 Primary

  Fungal source · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title

• C12N1/18

  Baker's yeast; Brewer's yeast · CPC title