# US 8,769,868 B2 · Compositions comprising eicosapentaenoic acid suitable for high purification

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- Number: US 8,769,868 B2
- Document ID: US8769868B2
- Status: Granted
- Kind: B2
- Application number: 13/702,616
- Filed: 2011-06-09
- Published: 2013-05-23
- Granted: 2014-07-08
- Earliest claimed: 2010-06-09
- Assignees: Photonz Corporation Limited
- Applicants: Hywel David Griffiths; Karl Thomas Geiringer; Mark Humphrey Dines
- Inventors: Hywel David Griffiths; Karl Thomas Geiringer; Mark Humphrey Dines
- Web page: https://patentgrep.com/patent/US8769868B2

## CPC

- C12N 1/00: Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N 1/12: Unicellular algae; Culture media therefor (as new plants A01H13/00)
- C12P 7/6432: Eicosapentaenoic acids [EPA]
- C12P 7/6458: by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
- 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)
- 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 relates to a microbial biomass composition produced from a heterotrophic fermentation whose fatty acid profile exhibits: an eicosapentaenoic acid (EPA) to arachidonic acid (ARA) ratio of about 11:1 or more; an EPA to total co-concentrating fatty acid ratio of about 8:1 or more, extract compositions, and methods of producing such compositions.

## Claims

### Claim 1

A method of producing a diatom, the method comprising:
a. cultivating diatoms in heterotrophic culture containing a nutrient solution comprising:
(i) a nitrogen source and an organic carbon source at an initial concentration of at least 0.5 M carbon,
(ii) a molar ratio of organic carbon to nitrogen of no higher than 30:1 (mol C:mol N), and
(iii) a non-limiting source of silicate at a concentration of 150 μM to 1.3 mM at all times during the culture; and
b. recovering the diatom from the heterotrophic culture,
wherein the cultivating conditions result in production of a diatom having at least 1.5% (w/w) of dry weight as eicosapentaenoic acid and an eicosapentaenoic acid to arachidonic acid ratio of at least 14:1.

### Claim 2 (depends on 1)

The method of claim 1 wherein the diatoms are cultivated in heterotrophic culture in a fermenter.

### Claim 3 (depends on 2)

The method of claim 2 wherein the cultivation comprises continuous fermentation.

### Claim 4 (depends on 1)

The method of claim 1 wherein the diatoms comprise Nitzschia species.

### Claim 5 (depends on 1)

The method of claim 1 wherein the cultivation is carried out at a temperature of from about 12° C. to about 35° C.

### Claim 6 (depends on 1)

The method of claim 1 wherein the cultivation is carried out at a pH of from about 7.0 to about 8.7.

### Claim 7 (depends on 1)

The method of claim 1 wherein said carbon source comprises glucose.

### Claim 8 (depends on 1)

The method of claim 1 wherein said carbon source is selected from glucose, hydrolysed starch or hydrolysed whey.

### Claim 9 (depends on 1)

The method of claim 1 wherein said nitrogen source is in the form of sodium nitrate or potassium nitrate.

### Claim 10 (depends on 1)

The method of claim 1 wherein said nitrogen sources is supplemented with an amino-acid source.

### Claim 11 (depends on 10)

The method of claim 10 wherein said amino-acid source is selected from corn-steep liquor, yeast extract, tryptone, peptones, lysine and glutamate.

### Claim 12 (depends on 1)

The method of claim 1 wherein said silicate is in the form of sodium or potassium metasilicate.

### Claim 13 (depends on 1)

The method of claim 1 wherein the nutrient solution comprises phosphorous in the form of phosphate.

### Claim 14 (depends on 13)

The method of claim 13 wherein the molar ratio of organic carbon to phosphorus in the form of phosphate is no higher than 1250:1 (mol C:mol P).