IP Library Granted Patent US 10,077,448
Granted Patent B2
US 10,077,448 · App. 15/044,408 · Granted Sep 18, 2018

Methods and systems for producing fungal secondary metabolites

Inventors: Nancy Keller (Madison, WI); Philipp Wiemann (Madison, WI)
Assignee: Wisconsin Alumni Research Foundation
C12N15/80C12P17/181
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Quick Facts
Patent No.
US 10,077,448
App. No.
15/044,408
Granted
Sep 18, 2018
Kind
B2
Abstract

The present invention discloses methods and systems for producing fungal secondary metabolites. The invention also discloses genetically modified organisms and kits including such organisms for producing fungal secondary metabolites.

Claims (20)

1. A genetically modified Aspergillus nidulans for producing one or more secondary metabolites comprising:

an Aspergillus nidulans organism comprising an engineered gene cluster comprising the A. nidulans sterigmatocystin gene cluster transcriptional enhancer genes aflR and aflJ operably linked to a nitrate-inducible promoter, wherein none of the other genes included in the wild type A. nidulans sterigmatocystin gene cluster are present in the engineered gene cluster;

wherein the genetically modified A. nidulans is capable of nitrate-induced expression of the aflR and aflJ gene products.

2. The genetically modified Aspergillus nidulans of claim 1 , wherein the wild type A. nidulans sterigmatocystin gene cluster is not present, and wherein the engineered gene cluster is inserted where the wild type A. nidulans sterigmatocystin gene cluster normally occurs.

3. The genetically modified Aspergillus nidulans of claim 1 , wherein the nitrate-inducible promoter is niiA/niaD.

4. The genetically modified Aspergillus nidulans of claim 1 , further comprising an exogenous expression vector comprising one or more A. nidulans sterigmatocystin gene cluster promoters operably linked to one or more protein-encoding genes, wherein the one or more A. nidulans sterigmatocystin gene cluster promoters are inducible by AflR/AflJ.

5. The genetically modified Aspergillus nidulans of claim 1 , wherein the genetically modified A. nidulans is strain TPMW2.3.

6. The genetically modified Aspergillus nidulans of claim 1 , wherein the nitrate-inducible promoter is repressible by ammonium.

7. The genetically modified Aspergillus nidulans of claim 4 , wherein the one or more protein-encoding genes are from a fungal secondary metabolite gene cluster, and wherein the genetically modified A. nidulans is capable of nitrate-induced expression of the secondary metabolite.

8. The genetically modified Aspergillus nidulans of claim 7 , wherein the exogenous expression vector comprises a fungal secondary metabolite gene cluster.

9. The genetically modified Aspergillus nidulans of claim 4 , wherein the proteins encoded by the protein-encoding genes are biosynthetic enzymes.

10. An expression vector for producing fungal secondary metabolites, comprising one or more Aspergillus nidulans sterigmatocystin gene cluster promoters operably linked to one or more protein-encoding genes that are not part of the A. nidulans sterigmatocystin gene cluster, wherein the one or more A. nidulans sterigmatocystin gene cluster promoters are inducible by AflR/AflJ.

11. The expression vector of claim 10 , comprising a fungal gene cluster other than the Aspergillus nidulans sterigmatocystin gene cluster.

12. The expression vector of claim 11 , wherein the fungal gene cluster is a secondary metabolite gene cluster.

13. A kit for producing fungal secondary metabolites, the kit comprising:

(a) the genetically modified Aspergillus nidulans of claim 1 ; and

(b) the expression vector of claim 10 .

14. A method for producing fungal secondary metabolites (SM), the method comprising contacting the genetically modified Aspergillus nidulans of claim 4 with nitrate, whereby one or more of the proteins encoded by the expression vector are expressed, and whereby a fungal secondary metabolite is produced.

15. The method of claim 14 , further comprising the step of producing the genetically modified Aspergillus nidulans of claim 4 by transforming the genetically modified A. nidulans of claim 1 with the expression vector of claim 10 .

16. The method of claim 14 , wherein the strain of Aspergillus nidulans is A. nidulans strain TPMW23.

Assignments (2)
CONFIRMATORY LICENSE Recorded May 24, 2018
From: UNIVERSITY OF WISCONSIN-MADISON
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 046237/0125 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 25, 2016
From: KELLER, NANCY; WIEMANN, PHILIPP
To: WISCONSIN ALUMNI RESEARCH FOUNDATION
Reel/Frame 037830/0423 →
Continuity (2)
Provisional Application 62116730 · Feb 16, 2015
Related Publication 20160237443A1 · Aug 18, 2016