# US 10,308,914 B2 · Processes for producing polyunsaturated fatty acids in transgenic organisms

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- Number: US 10,308,914 B2
- Document ID: US10308914B2
- Status: Granted
- Kind: B2
- Application number: 15/175,278
- Filed: 2016-06-07
- Published: 2016-10-20
- Granted: 2019-06-04
- Earliest claimed: 2006-10-06
- Assignees: BASF Plant Science GmbH
- Applicants: BASF Plant Science GmbH
- Inventors: Jörg Bauer; Tom Wetjen
- Web page: https://patentgrep.com/patent/US10308914B2

## CPC

- C12N 9/0071: {acting on paired donors with incorporation of molecular oxygen (1.14)}
- A23D 9/00: Other edible oils or fats, e.g. shortenings or cooking oils (preservation of other edible oils A23B20/30)
- A23D 9/02: characterised by the production or working-up
- A23K 20/158: Fatty acids; Fats; Products containing oils or fats
- A23K 50/80: for aquatic animals, e.g. fish, crustaceans or molluscs
- A23L 33/105: Plant extracts, their artificial duplicates or their derivatives
- A23L 33/115: Fatty acids or derivatives thereof; Fats or oils
- C12N 9/0083: {Miscellaneous (1.14.99)}
- C12N 9/1029: {transferring groups other than amino-acyl groups (2.3.1)}
- C12N 9/93: {Ligases (6)}
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- C12P 7/6427: Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P 7/6432: Eicosapentaenoic acids [EPA]
- C12P 7/6434: Docosahexenoic acids [DHA]
- C12P 7/6472: containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
- A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C12Y 114/19001: Stearoyl-CoA 9-desaturase (1.14.19.1), i.e. DELTA9-desaturase
- C12Y 602/01003: Long-chain-fatty-acid-CoA ligase (6.2.1.3)
- Y02A 40/818: Alternative feeds for fish, e.g. in aquacultures
- Y02P 20/52: using catalysts, e.g. selective catalysts
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A23D: EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS (preservation thereof A23B20/00; production, refinement or preservation of animal or vegetable fats or oils C11B, C11C)
- A23K: FODDER
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- 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)}
- A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12Y: ENZYMES
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The present invention relates to polynucleotides from Ostreococcus lucimarinus which code for desaturases and elongases and which can be employed for the recombinant production of polyunsaturated fatty acids. The invention furthermore relates to vectors, host cells and transgenic nonhuman organisms which comprise the polynucleotides, and to the polypeptides encoded by the polynucleotides. Finally, the invention also relates to production processes for the polyunsaturated fatty acids and for oil, lipid and fatty acid compositions.

## Claims

### Claim 1

A polynucleotide comprising an expression control sequence operably linked to a heterologous nucleic acid sequence selected from the group consisting of:
a) the nucleic acid sequence of SEQ ID NO: 9;
b) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 10;
c) a nucleic acid sequence having at least 85% sequence identity to the nucleic acid sequence of a) or b); and
d) a nucleic acid sequence encoding a polypeptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 10,
wherein said heterologous nucleic acid sequence encodes a polypeptide having desaturase activity.

### Claim 2 (depends on 1)

The polynucleotide of claim 1, wherein said polynucleotide further comprises a terminator sequence operably linked to said heterologous nucleic acid sequence.

### Claim 3 (depends on 1)

A vector comprising the polynucleotide of claim 1.

### Claim 4 (depends on 1)

A host cell comprising:
a) the polynucleotide of claim 1; or
b) a vector comprising said polynucleotide of claim 1.

### Claim 5 (depends on 4)

A method for the manufacture of polyunsaturated fatty acids, comprising:
a) cultivating the host cell of claim 4 under conditions which allow for the production of polyunsaturated fatty acids in said host cell; and
b) obtaining said polyunsaturated fatty acids from said host cell.

### Claim 6 (depends on 5)

The method of claim 5, wherein said polyunsaturated fatty acid is arachidonic acid (ARA), eicosapentaenoic acid (EPA), and/or docosahexaenoic acid (DHA).

### Claim 7 (depends on 4)

A method for the manufacture of an oil, lipid, or fatty acid composition, comprising:
a) cultivating the host cell of claim 4 under conditions which allow for the production of polyunsaturated fatty acids in said host cell;
b) obtaining said polyunsaturated fatty acids from said host cell; and
c) formulating the polyunsaturated fatty acid as an oil, lipid, or fatty acid composition.

### Claim 8 (depends on 7)

The method of claim 7, further comprising:
processing said oil-, lipid-, or fatty acid composition to produce feed, foodstuffs, cosmetics or pharmaceuticals.

### Claim 9 (depends on 1)

The polynucleotide of claim 1, wherein the nucleic acid sequence of c) has at least 90% sequence identity to the nucleic acid sequence of a) or b).

### Claim 10 (depends on 1)

The polynucleotide of claim 1, wherein the nucleic acid sequence of c) has at least 95% sequence identity to the nucleic acid sequence of a) or b).

### Claim 11 (depends on 1)

The polynucleotide of claim 1, wherein the nucleic acid sequence of d) encodes a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 10.

### Claim 12 (depends on 1)

The polynucleotide of claim 1, wherein the nucleic acid sequence of d) encodes a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 10.

### Claim 13 (depends on 4)

The host cell of claim 4, wherein said host cell is a non-human animal cell or a plant cell.

### Claim 14 (depends on 5)

The method of claim 5, wherein said host cell is a non-human animal cell or a plant cell.

### Claim 15 (depends on 7)

The method of claim 7, wherein said host cell is a non-human animal cell or a plant cell.