# US 9,752,134 B2 · Polypeptides having glucoamylase activity and polynucleotides encoding same

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- Number: US 9,752,134 B2
- Document ID: US9752134B2
- Status: Granted
- Kind: B2
- Application number: 14/934,911
- Filed: 2015-11-06
- Published: 2016-02-25
- Granted: 2017-09-05
- Earliest claimed: 2009-12-01
- Assignees: Novozymes North America, Inc.; Novozymes A/S
- Applicants: Novozymes A/S; Novozymes North America, Inc.
- Inventors: Sara Landvik; Marc Dominique Morant; Keiichi Ayabe; Guillermo Coward-Kelly
- Web page: https://patentgrep.com/patent/US9752134B2

## CPC

- C12N 9/2428: {Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase}
- C12N 15/81: for yeasts
- C12P 7/10: substrate containing cellulosic material
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C12Y 302/01003: Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The present invention relates to polypeptides having glucoamylase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

## Claims

### Claim 1

A recombinant host cell comprising a nucleic acid construct comprising a polynucleotide encoding a glucoamylase according to the invention operably linked to one or more control sequences that direct the production of the polypeptide in the expression host, wherein the host cell is a yeast cell and wherein the glucoamylase is selected from the group consisting of:
a polypeptide having glucoamylase activity, having at least 90% identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, or SEQ ID NO: 18.

### Claim 2

A recombinant host cell comprising a nucleic acid construct comprising a polynucleotide encoding a glucoamylase comprising a catalytic domain according to the invention operably linked to one or more control sequences that direct the production of the glucoamylase comprising a catalytic domain polypeptide in the expression host, wherein the host cell is a yeast cell and wherein the glucoamylase catalytic domain is selected from the group consisting of:
a polypeptide having at least 90% identity, to the sequence of amino acids 19 to 474 of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or amino acids 19 to 471 of SEQ ID NO: 16 or SEQ ID NO: 18.

### Claim 3 (depends on 1)

The recombinant host cell according to claim 1, wherein the yeast host cell is a Candida, Hansenula, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia cell.

### Claim 4 (depends on 3)

The recombinant host cell according to claim 3, wherein the yeast host cell is a Saccharomyces cell.

### Claim 5 (depends on 4)

The recombinant host cell according to claim 4, wherein the yeast host cell is a Saccharomyces carlsbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri, Saccharomyces norbensis, or Saccharomyces oviformis cell.

### Claim 6 (depends on 5)

The recombinant host cell according to claim 5, wherein the yeast host cell is a Saccharomyces cerevisiae cell.

### Claim 7 (depends on 6)

The recombinant host cell according to claim 6, wherein the polypeptide having glucoamylase activity is selected from SEQ ID NO: 14 or a polypeptide having at least 90% identity to the mature polypeptide of SEQ ID NO: 14.

### Claim 8 (depends on 6)

The recombinant host cell according to claim 6, wherein the glucoamylase catalytic domain is selected from amino acids 19 to 474 SEQ ID NO: 14 or a polypeptide having at least 90% identity to amino acids 19 to 474 of SEQ ID NO: 14.