# US 2019/0169584 A1 · Promoter useful for high expression of a heterologous gene of interest in Aspergillus niger

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- Number: US 2019/0169584 A1
- Document ID: US20190169584A1
- Status: Published application
- Kind: A1
- Application number: 16/124,127
- Filed: 2018-09-06
- Published: 2019-06-06
- Earliest claimed: 2017-09-06
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California; National Technology & Engineering Solutions of Sandia, LLC; Pacific Northwest National Laboratory (PNNL)
- Inventors: John M. Gladden; Saori Amaike Campen; Jinxiang Zhang; Jon K. Magnuson; Scott E. Baker; Blake A. Simmons
- Web page: https://patentgrep.com/patent/US20190169584A1

## CPC

- C12N 9/242: {Fungal source}
- C07K 14/38: from Aspergillus
- C12N 9/2445: {Beta-glucosidase (3.2.1.21)}
- C12N 15/80: for fungi
- C12Y 302/0102: Alpha-glucosidase (3.2.1.20)
- C12Y 302/01021: Beta-glucosidase (3.2.1.21)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12Y: ENZYMES

## Abstract

The present invention provides for an Asperigillus niger host cell comprising a gene of interest operatively linked to an ecm33 promoter of an ascomycete fungi, wherein the gene of interest is heterologous to the ecm33 promoter and/or to Asperigillus niger. In some embodiments, the gene of interest is a glycoside hydrolase enzyme. In some embodiments, the glycoside hydrolase enzyme is a glucosidase.

## Claims

### Claim 1

A nucleic acid encoding a gene of interest operatively linked to an ecm33 promoter of an ascomycete fungi, wherein the gene of interest is heterologous to the ecm33 promoter and/or to Asperigillus niger.

### Claim 2 (depends on 1)

The nucleic acid of claim 1, wherein the ascomycete fungi is an Asperigillus species.

### Claim 3 (depends on 2)

The nucleic acid of claim 2, wherein the Asperigillus species is an Asperigillus niger.

### Claim 4 (depends on 3)

The nucleic acid of claim 3, wherein the Asperigillus niger is an Asperigillus niger strain 1015.

### Claim 5 (depends on 1)

The nucleic acid of claim 1, wherein the ecm33 promoter comprises the nucleotide sequence of SEQ ID NO:1.

### Claim 6 (depends on 1)

The nucleic acid of claim 1, wherein the gene of interest is a glycoside hydrolase enzyme.

### Claim 7 (depends on 6)

The nucleic acid of claim 6, wherein the glycoside hydrolase enzyme is a glucosidase.

### Claim 8 (depends on 1)

A vector comprising the nucleic acid of claim 1.

### Claim 9 (depends on 8)

The vector of claim 8, wherein the vector is an expression vector.

### Claim 10 (depends on 8)

The vector of claim 8, wherein the gene of interest is a glycoside hydrolase enzyme.

### Claim 11 (depends on 10)

The vector of claim 10, wherein the glycoside hydrolase enzyme is a glucosidase.

### Claim 12

An Asperigillus niger host cell comprising a gene of interest operatively linked to an ecm33 promoter of an ascomycete fungi, wherein the gene of interest is heterologous to Asperigillus niger.

### Claim 13 (depends on 1)

An Asperigillus niger host cell comprising the nucleic acid of claim 1.

### Claim 14 (depends on 1)

A method of expressing a heterologous gene of interest in an Asperigillus niger host cell, comprising: (a) optionally constructing the nucleic acid of claim 1, (b) introducing the nucleic acid into an A. niger host cell, (c) culturing or growing the host cell in a medium suitable for expressing the gene of interest, and (d) optionally separating or purifying the gene product encoded by the gene of interest from the rest of the host cell and/or medium.