# US 9,040,730 B2 · Purification of triglyceride oil from microbial sources using short path distillation

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- Number: US 9,040,730 B2
- Document ID: US9040730B2
- Status: Granted
- Kind: B2
- Application number: 13/370,785
- Filed: 2012-02-10
- Published: 2013-02-21
- Granted: 2015-05-26
- Earliest claimed: 2011-02-11
- Assignees: E. I. Du Pont de Nemours and Company
- Applicants: Shu-Chien Liang; Robert D Orlandi
- Inventors: Shu-Chien Liang; Robert D Orlandi
- Web page: https://patentgrep.com/patent/US9040730B2

## CPC

- B01D 3/12: Molecular distillation
- A23L 33/115: Fatty acids or derivatives thereof; Fats or oils
- C11B 1/104: {using super critical gases or vapours}
- C11B 3/12: by distillation
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- C11B: PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES (drying-oils C09F)

## Abstract

Disclosed is a process for reducing the amount of sterol in a sterol-containing microbial oil composition, including distilling, under short path distillation conditions, a sterol-containing microbial oil wherein said distillation produces a distillate fraction containing the sterol and a triacylglycerol-containing fraction having a reduced amount of the sterol when compared to the amount of sterol in the sterol-containing microbial oil composition that has not been subjected to short path distillation.

## Claims

### Claim 1

A process for reducing the amount of sterol in a sterol-containing microbial oil composition obtained from microbial biomass of recombinant Yarrowia cells, said method comprising:
a) distilling, at least once under short path distillation conditions, the sterol-containing microbial oil composition, wherein said oil composition comprises:
(i) triacylglycerol comprising one or more polyunsaturated fatty acids; and
(ii) a sterol fraction of at least 300 mg/100 g of oil, wherein the sterol fraction comprises one or more sterols selected from the group consisting of stigmasterol, ergosterol, brassicasterol, campesterol, beta-sitosterol, and desmosterol;
wherein said distillation produces a distillate fraction comprising the sterol and a triacylglycerol-containing fraction having a reduced amount of the sterol when compared to the amount of sterol in the sterol-containing microbial oil composition that has not been subjected to short path distillation; and
b) optionally, recovering the triacylglycerol-containing fraction;
wherein said Yarrowia cells are engineered for production of eicosapentaenoic acid and at least 20% of the dry cell weight of said Yarrowia cells is eicosapentaenoic acid.

### Claim 2 (depends on 1)

The process of claim 1, wherein the short path distillation conditions comprise at least one pass of the sterol-containing microbial oil composition at a vacuum level of not more than 30 mTorr and a temperature of not more than 300° C.

### Claim 3 (depends on 1)

The process of claim 1, wherein the sterol fraction comprises ergosterol.

### Claim 4 (depends on 1)

The process of claim 1, wherein the reduction in the amount of sterols in the triacylglycerol-containing fraction is at least 40% when compared to the amount of sterols in the sterol-containing microbial oil composition.

### Claim 5 (depends on 1)

The process of claim 1, wherein the triacylglycerol-containing fraction has improved clarity when compared to the sterol-containing microbial oil composition that has not been subjected to short path distillation.

### Claim 6 (depends on 2)

The process of claim 2, wherein the temperature is not more than 280° C.

### Claim 7 (depends on 1)

The process of claim 1, wherein the sterol-containing microbial oil composition is a refined lipid composition having less than 20 ppm phosphorous as determined with inductively coupled plasma optical emission spectroscopy.

### Claim 8 (depends on 1)

The process of claim 1, wherein the recombinant Yarrowia cells are further engineered for production of at least one polyunsaturated fatty acid selected from the group consisting of: linoleic acid, gamma-linolenic acid, eicosadienoic acid, dihomo-gamma-linolenic acid, arachidonic acid, docosatetraenoic acid, omega-6 docosapentaenoic acid, alpha-linolenic acid, stearidonic acid, eicosatrienoic acid, eicosatetraenoic acid, omega-3 docosapentaenoic acid, docosahexaenoic acid, and mixtures thereof.

### Claim 9 (depends on 1)

The process of claim 1, wherein said distilling comprises two or more consecutive short path distillations of the microbial oil composition.

### Claim 10 (depends on 9)

The process of claim 9, wherein each consecutive short path distillation is at a temperature that is higher than the temperature of the immediately preceding short path distillation.