# US 10,119,153 B2 · Method for arachidonic acid production from salispina spinosa (halophytophthora spinosa)

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- Number: US 10,119,153 B2
- Document ID: US10119153B2
- Status: Granted
- Kind: B2
- Application number: 14/582,313
- Filed: 2014-12-24
- Published: 2015-07-02
- Granted: 2018-11-06
- Earliest claimed: 2013-12-26
- Assignees: National Taiwan Ocean University
- Applicants: National Taiwan Ocean University
- Inventors: Ka-Lai Pang; Han-Jia Lin
- Web page: https://patentgrep.com/patent/US10119153B2

## CPC

- C12P 7/6427: Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12N 1/14: Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor
- C12P 7/6432: Eicosapentaenoic acids [EPA]
- C12N 1/145: Fungi isolates
- C12R 2001/645: Fungi {; Processes using fungi}
- 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)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention provides a method for eicosapentaenoic acid (EPA) and/or arachidonic acid (ARA) production by using Halophytophthora and Halophytophthora isolates that are suitable for EPA and/or ARA production. The disclosure of the present invention provides another choice for the industry to produce EPA and ARA that are good for human's health and promotes the industrial value of Halophytophthora.

## Claims

### Claim 1

A method for arachidonic acid production, comprising isolating arachidonic acid from a Salispina spinosa strain,
wherein said Salispina spinosa strain has a deposition number of DSM 32804 at the Leibniz Institute DSMZ—German Collection of Microorganisms and Cell Cultures.

### Claim 2 (depends on 1)

The method of claim 1, further comprising a culture step before conducting said isolating; wherein said culture step comprises:
culturing said Salispina spinosa strain in a liquid medium for 4 to 10 days; wherein said medium is PYGS medium, Cornmeal medium, Potato Dextrose medium, or a combination thereof.

### Claim 3 (depends on 1)

The method of claim 1, further comprising a culture step before conducting said isolating; wherein said culture step comprises:
culturing said Salispina spinosa strain in a solid medium for 4 to 10 days to obtain a Salispina spinosa strain colony; and
culturing said colony in a liquid medium for 4 to 10 days;
wherein said medium is PYGS medium, Cornmeal medium, Potato Dextrose medium, or a combination thereof.

### Claim 4 (depends on 3)

The method of claim 3, wherein said medium comprises 1.0 to 3.5 wt % of salt; wherein said wt % is based on the total weight of said medium.

### Claim 5 (depends on 3)

The method of claim 3, wherein said medium has a pH value of 5 to 9.

### Claim 6 (depends on 3)

The method of claim 3, wherein said Salispina spinosa strain is cultured at a temperature of 10 to 35° C.

### Claim 7 (depends on 3)

The method of claim 3, wherein said PYGS medium comprises:
1 to 5 g/L of glucose;
1 to 5 g/L of yeast extract;
1 to 5 g/L of peptone; and
1 to 3.5% (v/v) of seawater;
Wherein said unit of concentration is based on the total volume of said PYGS medium.

### Claim 8 (depends on 3)

The method of claim 3, wherein the yield of said arachidonic acid is 0.05 to 9 mg/L/day; wherein said day is calculated from the sum of the days that said Salispina spinosa strain is cultured in said liquid medium.

### Claim 9 (depends on 7)

The method of claim 7, wherein said PYGS medium comprises:
1 to 5 g/L of glucose;
1 to 5 g/L of yeast extract;
1 to 5 g/L of peptone; and
2 to 3% (v/v) of seawater;
wherein said unit of concentration is based on the total volume of said PYGS medium.