Method for manufacturing useful substance in which high-density cultured strain of filamentous fungi is used

Patent metadata

Field	Value
Publication number	US-11015175-B2
Application number	US-201314441493-A
Country	US
Kind code	B2
Filing date	Nov 8, 2013
Priority date	Nov 9, 2012
Publication date	May 25, 2021
Grant date	May 25, 2021

Abstract

An object to be achieved by the present invention is to culture a filamentous fungus at a high density, thereby enabling mass production of a useful substance. The present invention provides a method of producing a substance, including the steps of: culturing a mutant filamentous fungus with no expression of α-1,3-glucan to allow the filamentous fungus to produce a substance; and collecting the resulting substance.

First claim

The invention claimed is:

1. A method for increasing the production amount of protein per unit volume in a mutant filamentous fungus which lacks α-1,3-glucan compared to the corresponding parent filamentous fungus comprising the steps of:
culturing the mutant filamentous fungus to allow said fungus to undergo a hyphal growth and produce the protein; and collecting the produced protein,
wherein the mutant filamentous fungus is a mutant Aspergillus oryzae, and wherein the mutant filamentous fungus is deficient in agsB gene encoding α-1,3-glucan synthase and further deficient in at least one α-1,3-glucan synthase ags gene other than the agsB gene, and wherein the culturing step is conducted in liquid culture medium.

2. A method according to claim 1 , wherein the protein is selected from the group consisting of amylase, cellulase, protease, lipase, peptidase, esterase, and oxidase.

3. The method according to claim 1 , wherein a culture time of the mutant filamentous fungus is more than 24 hours in the culturing step.

4. The method according to claim 1 , wherein the mutant filamentous fungus is cultured to the vegetative growth stage in the culturing step.

5. The method according to claim 1 , wherein the culturing step is conducted for 12 hours or more.

6. A method for increasing the density of hyphae of a mutant filamentous fungus which lacks α-1,3-glucan compared to the corresponding parent filamentous fungus comprising the steps of:
culturing the mutant filamentous fungus to allow for said fungus undergoing a hyphal growth to reach the increased level of the density of hyphae which is associated with protein production in said mutant filamentous fungus and
collecting the produced protein,
wherein the mutant filamentous fungus is a mutant Aspergillus oryzae, and wherein the mutant filamentous fungus is deficient in agsB gene encoding α-1,3-glucan and further deficient in at least one α-1,3-glucan synthase ags gene other than the agsB gene, and wherein the culturing step is conducted in liquid culture medium.

7. The method according to claim 6 , wherein a culture time of the mutant filamentous fungus is more than 24 hours in the culturing step.

8. The method according to claim 6 , wherein the mutant filamentous fungus is cultured to the vegetative growth stage in the culturing step.

9. The method according to claim 6 , wherein the culturing step is conducted for 12 hours or more.

Assignees

• Univ Tohoku

Inventors

• Abe Keietsu
• Gomi Katsuya
• Yoshimi Kei

Classifications

• C12N9/485

  Exopeptidases (3.4.11-3.4.19) · CPC title

• C12P35/00

  Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system, e.g. cephalosporin · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12P1/02

  by using fungi · CPC title

• C12N9/2411

  Amylases · CPC title