Production of 2-keto-3-deoxy-d-gluconic acid in filamentous fungi

Patent metadata

Field	Value
Publication number	US-11708571-B2
Application number	US-201917056146-A
Country	US
Kind code	B2
Filing date	May 14, 2019
Priority date	May 17, 2018
Publication date	Jul 25, 2023
Grant date	Jul 25, 2023

Abstract

A recombinant filamentous fungi that includes reduced 2-Keto-3-Deoxy-Gluconate (KDG) aldolase enzyme activity as compared to the filamentous fungi not transformed to have reduced KDG aldolase enzyme activity is provided. Also provided is a method of producing KDG.

First claim

What is claimed is:

1. A genetically modified filamentous fungus having reduced 2-Keto-3-Deoxy-Gluconate (KDG) aldolase enzyme activity as compared to the KDG aldolase enzyme activity in a wild type counterpart of said filamentous fungus, wherein said filamentous fungus is an Aspergillus niger strain.

2. The filamentous fungus claim of 1 , wherein the Aspergillus niger strain is selected from the group consisting of NRRL 322, NRRL 328, NRRL 566, NRRL 599 and NRRL 2270.

3. The filamentous fungus of claim 1 , wherein the genetic modification results in reduced endogenous KDG aldolase enzyme activity.

4. The filamentous fungus of claim 1 , wherein the genetic modification is a mutation.

5. The filamentous fungus of claim 4 , wherein the mutation is selected from the group consisting of a frame shift mutation, a substitution mutation, an insertion mutation, a loss of function mutation, a gain of function mutation, an inactivation mutation, a translocation mutation and a deletion mutation.

6. The filamentous fungus of claim 4 , wherein the mutation is in the promotor region, the 3′ untranslated region, the 5′ untranslated region and/or a regulatory sequence of a 2-Keto-3-Deoxy-Gluconate (KDG) aldolase gene.

7. The filamentous fungus of claim 4 , wherein the mutation is in a nucleic acid sequence encoding a polypeptide having complete sequence identity to SEQ ID NO:1.

8. The filamentous fungus of claim 4 , wherein the mutation is a deletion of a gene having the nucleic acid sequence of SEQ ID NO:2.

9. The filamentous fungus of claim 4 , wherein the mutation is in a nucleic acid sequence encoding a KDG aldolase enzyme comprising the amino acid sequence of SEQ ID NO:1.

10. The filamentous fungus of claim 4 , wherein the mutation inhibits expression of the polypeptide comprising SEQ ID NO:1.

11. The filamentous fungus of claim 1 , wherein the enzyme having reduced 2-Keto-3-Deoxy-Gluconate (KDG) aldolase enzyme activity has the classification of EC4.1.2.51.

12. A method of producing 2-Keto-3-Deoxy-Gluconate (KDG) or a furan derivative comprising:
culturing the genetically modified filamentous fungus of claim 1 in a culture medium, wherein the level of KDG or a furan derivative accumulated by the genetically modified filamentous fungus is higher than the level of KDG or a furan derivative accumulated by a wild type counterpart of said filamentous fungus, thereby producing KDG or a furan derivative.

13. The method of claim 12 , wherein the genetically modified filamentous fungus has been transformed with a nucleic acid sequence that inactivates a gene encoding a KDG aldolase enzyme, reduces expression of a gene encoding a KDG aldolase enzyme or inhibits expression of a gene encoding a KDG aldolase enzyme.

14. A method of decreasing D-glyceraldehyde production from 2-Keto-3-Deoxy-Gluconate in a filamentous fungus comprising:
culturing the genetically modified filamentous fungus of claim 1 in a culture medium, wherein the level of D-glyceraldehyde production from 2-Keto-3-Deoxy-Gluconate in the genetically modified filamentous fungus is decreased from the level of glyceraldehyde production from 2-Keto-3-Deoxy-Gluconate in a wild type counterpart of said filamentous fungus, thereby decreasing D-glyceraldehyde from 2-Keto-3-Deoxy-Gluconate.

15. The method of claim 14 , wherein the genetically modified filamentous fungus has been transformed with a nucleic acid sequence that inactivates a gene encoding a KDG aldolase enzyme, reduces expression of a gene encoding a KDG aldolase enzyme or inhibits expression of a gene encoding a KDG aldolase enzyme.

Assignees

• Bp Corp North America Inc

Inventors

• Krebs Molly
• Phillips Chris
• Valle Fernando

Classifications

• C12N9/88 Primary

  Lyases (4.) · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title

• C12P7/42

  Hydroxy-carboxylic acids · CPC title

• C12Y401/02014

  2-Dehydro-3-deoxy-phosphogluconate aldolase (4.1.2.14) · CPC title

• C12P17/04

  containing a five-membered hetero ring, e.g. griseofulvin {, vitamin C} · CPC title