# US 8,541,210 B2 · Producing eicosapentaenoic acid (EPA) from biodiesel-derived crude glycerol

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- Number: US 8,541,210 B2
- Document ID: US8541210B2
- Status: Granted
- Kind: B2
- Application number: 13/486,464
- Filed: 2012-06-01
- Published: 2013-01-03
- Granted: 2013-09-24
- Earliest claimed: 2008-05-19
- Assignees: Virginia Tech Intellectual Properties, Inc.
- Applicants: Zhiyou Wen; Sneba Athalye
- Inventors: Zhiyou Wen; Sneba Athalye
- Web page: https://patentgrep.com/patent/US8541210B2

## CPC

- C12P 7/6472: containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
- C12P 7/6432: Eicosapentaenoic acids [EPA]
- C12P 7/6458: by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
- 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

Pythium, when cultured with crude glycerol as a carbon source, produces useful polyunsaturated fatty acids such as eicosapentaenoic acid (EPA). The crude glycerol is pretreated to remove soaps and methanol. An exemplary Pythium species for use in the production of EPA is Pythium irregulare.

## Claims

### Claim 1

A method of producing an eicosapentaenoic acid-rich biomass from crude waste glycerol, the method comprising the steps of:
providing crude waste glycerol in a culture medium; and
culturing Pythium in the culture medium under conditions that permit the Pythium to use the crude glycerol in the medium as a carbon source to yield a eicosapentaenoic acid-rich biomass,
wherein the Pythium eicosapentaenoic acid-rich biomass has a total lipids content of at least about 15 weight percent and the crude glycerol is substantially free of soaps and methanol.

### Claim 2 (depends on 1)

The method of claim 1, wherein the Pythium biomass is Pythium irregulare.

### Claim 3 (depends on 1)

The method of claim 1, wherein the step of providing the crude glycerol includes a step of removing soaps and/or methanol from the crude glycerol.

### Claim 4 (depends on 1)

The method of claim 1, wherein the crude glycerol culture further comprises one or more oils.

### Claim 5 (depends on 1)

The method of claim 1, wherein the one or more oils are selected from flaxseed oil, soybean oil, palm oil, rapeseed oil, sunflower seed oil, peanut oil, cottonseed oil, coconut oil, olive oil.

### Claim 6 (depends on 1)

The method of claim 1, wherein the crude glycerol is present in the culture medium at a concentration of at least 30 grams per liter.

### Claim 7 (depends on 1)

The method of claim 1, wherein the culture medium further comprises 10 grams per liter or more of yeast extract.

### Claim 8 (depends on 1)

The method of claim 1, wherein the culture medium is pretreated by removing soaps and/or methanol from the crude glycerol.

### Claim 9 (depends on 1)

The method of claim 1, wherein the Pythium biomass comprises at least 7 percent eicosapentaenoic acid in the total lipids portion of the biomass.

### Claim 10 (depends on 1)

The method of claim 1, wherein the Pythium biomass is grown to a cell density of at least 6 grams per liter.