Methods and systems for producing fungal secondary metabolites

Patent metadata

Field	Value
Publication number	US-2016237443-A1
Application number	US-201615044408-A
Country	US
Kind code	A1
Filing date	Feb 16, 2016
Priority date	Feb 16, 2015
Publication date	Aug 18, 2016
Grant date	—

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.

First claim

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 TPMW2.3.

Assignees

• Wisconsin Alumni Res Found

Inventors

• Keller Nancy
• Wiemann Philipp

Classifications

• C12P17/181

  Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin · CPC title

• C12N15/80 Primary

  for fungi · CPC title

• C12P17/06

  containing a six-membered hetero ring, e.g. fluorescein · CPC title

• C12P37/00

  Preparation of compounds having a 4-thia-1-azabicyclo [3.2.0] heptane ring system, e.g. penicillin · CPC title

• C12P1/02

  by using fungi · CPC title