# US 7,608,703 B2 · PUFA polyketide synthase systems and uses thereof

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- Number: US 7,608,703 B2
- Document ID: US7608703B2
- Status: Granted
- Kind: B2
- Application number: 11/689,605
- Filed: 2007-03-22
- Published: 2008-02-14
- Granted: 2009-10-27
- Earliest claimed: 1999-01-14
- Assignees: Martek Biosciences Corporation
- Inventors: James G. Metz; James H. Flatt; Jerry M. Kuner; William R. Barclay
- Web page: https://patentgrep.com/patent/US7608703B2

## CPC

- C12N 15/52: Genes encoding for enzymes or proenzymes
- C07K 14/28: from Vibrionaceae (F)
- C07K 14/405: from algae
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- C12N 15/8273: {for drought, cold, salt resistance}
- C12P 7/625: Polyesters of hydroxy carboxylic acids
- 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
- 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)
- 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)}

## Abstract

The invention generally relates to polyunsaturated fatty acid (PUFA) polyketide synthase (PKS) systems isolated from or derived from non-bacterial organisms, to homologues thereof, to isolated nucleic acid molecules and recombinant nucleic acid molecules encoding biologically active domains of such a PUFA PKS system, to genetically modified organisms comprising PUFA PKS systems, to methods of making and using such systems for the production of bioactive molecules of interest, and to novel methods for identifying new bacterial and non-bacterial microorganisms having such a PUFA PKS system.

## Claims

### Claim 1

An isolated nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to SEQ ID NO:24 or that is an enzymatically active fragment of SEQ ID NO:24, wherein said amino acid sequence has acyl transferase activity.

### Claim 2 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to SEQ ID NO:24, wherein said amino acid sequence has acyl transferase activity.

### Claim 3 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is an enzymatically active fragment of SEQ ID NO:24, wherein said amino acid sequence has acyl transferase activity.

### Claim 4 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to SEQ ID NO:24, wherein said amino acid sequence acyl transferase activity.

### Claim 5 (depends on 1)

The isolated nucleic acid molecule of claim 1, consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to SEQ ID NO:24, wherein said amino acid sequence has acyl transferase activity.

### Claim 6 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is at least 96% identical to SEQ ID NO:24 wherein said amino acid sequence has acyl transferase activity.

### Claim 7 (depends on 1)

The isolated nucleic acid molecule of claim 1, consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 96% identical to SEQ ID NO:24 wherein said amino acid sequence has acyl transferase activity.

### Claim 8 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is at least 97% identical to SEQ ID NO:24 wherein said amino acid sequence has acyl transferase activity.

### Claim 9 (depends on 1)

The isolated nucleic acid molecule of claim 1, consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 97% identical to SEQ ID NO:24 wherein said amino acid sequence has acyl transferase activity.

### Claim 10 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is at least 98% identical to SEQ ID NO:24 wherein said amino acid sequence has acyl transferase activity.

### Claim 11 (depends on 1)

The isolated nucleic acid molecule of claim 1, consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 98% identical to SEQ ID NO:24 wherein said amino acid sequence has acyl transferase activity.

### Claim 12 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding an amino acid sequence that is at least 99% identical to SEQ ID NO:24, wherein said amino acid sequence has acyl transferase activity.

### Claim 13 (depends on 1)

The isolated nucleic acid molecule of claim 1, consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 99% identical to SEQ ID NO:24, wherein said amino acid sequence has acyl transferase activity.

### Claim 14 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:24.

### Claim 15 (depends on 1)

The isolated nucleic acid molecule of claim 1, consisting of a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:24.

### Claim 16 (depends on 1)

A recombinant nucleic acid molecule comprising the nucleic acid molecule of claim 1 and a transcription control sequence.

### Claim 17 (depends on 1)

A recombinant plant cell that expresses the nucleic acid molecule of claim 1.

### Claim 18 (depends on 1)

A recombinant microbial cell that expresses a recombinant vector comprising the nucleic acid molecule of claim 1 and a transcription control sequence.

### Claim 19 (depends on 18)

The recombinant microbial cell of claim 18, wherein the microbial cell is a bacterium.

### Claim 20 (depends on 18)

The recombinant microbial cell of claim 18, wherein the microbial cell is a Thraustochytriales microorganism.

### Claim 21 (depends on 20)

The recombinant microbial cell of claim 20, wherein the Thraustochytriales microorganism is a Schizochytrium or a Thraustochytrium.

### Claim 22 (depends on 1)

A method to produce at least one polyunsaturated fatty acid (PUFA), comprising culturing under conditions effective to produce the PUFA, a plant cell or a microbial cell that expresses a PKS system for production of PUFAs, wherein the plant cell or microbial cell expresses a recombinant vector comprising the nucleic acid molecule of claim 1.
