Preparation of microbial oil

Patent metadata

Field	Value
Publication number	US-10633678-B2
Application number	US-201314027588-A
Country	US
Kind code	B2
Filing date	Sep 16, 2013
Priority date	Jun 19, 2002
Publication date	Apr 28, 2020
Grant date	Apr 28, 2020

Abstract

The present invention provides a process for the production of a microbial oil comprising culturing a micro-organism in a two stage fermentation process where, in a last stage that precedes the end of fermentation, the carbon source is: consumed by the micro-organisms at a rate greater than it is added to the medium; added at a rate #0.30 M carbon/kg medium; or is rate limiting on the growth of the micro-organism. The micro-organisms thus have the carbon source restricted so that they preferentially metabolise fats or lipids other than arachidonic acid (ARA), so increasing the proportion of ARA in the cells. A microbial oil is then recovered from the micro-organism, using hexane as a solvent, that has at least 50% ARA and at least 90% triglycerides.

First claim

We claim:

1. A fermentation process for production of a microbial oil comprising at least 40% arachidonic acid (ARA), the process comprising culturing a microorganism which is Mortierella alpina in a two-stage process in a culture medium inside a fermentation vessel, whereby at a second stage of the fermentation process
(a) a carbon source is rate limiting on the growth of the microorganism,
(b) the carbon source is restricted such that the microorganism metabolises fat(s) and/or lipid(s) in preference to arachidonic acid (ARA),
(c) the carbon source is consumed by the microorganism at a rate greater than it is added to the medium,
(d) the carbon source is added at an addition rate of 0.5 g/kg/hr, and
(e) the carbon source is controlled so that the concentration of the carbon source is about 5 g/kg of medium about 10 hours before the end of fermentation;
whereby the process further comprises extracting the oil from the micro-organism using a non-polar solvent, and wherein the oil has a triglyceride content of at least 90%.

2. The process according to claim 1 , wherein:
concentration of the carbon source in the second stage is on average ≤0.17M carbon/kg medium.

3. The process according to claim 1 , wherein
fermentation is carried out at a temperature ≥22° C. and ≤30° C.

4. The process according to claim 1 , wherein the carbon source is glucose.

5. The process according to claim 1 , wherein during an earlier stage of fermentation, the carbon source is being added, wherein the earlier stage is prior to the second stage.

6. The process according to claim 5 , wherein during said earlier stage the carbon source is in excess.

7. The process according to claim 1 , wherein cells of the microorganism are pasteurized after fermentation is finished.

8. The process according to claim 1 , wherein the oil comprises at least 50% ARA (wt/wt) based on the oil.

9. The process according to claim 1 , wherein a concentration of the carbon source in said second stage is <4 g/kg of medium.

10. The process according to claim 1 , wherein the solvent is hexane.

11. The process according to claim 8 , wherein:
concentration of the carbon source in the second stage is on average ≤0.17 M carbon/kg medium.

12. The process according to claim 8 , wherein fermentation is carried out at a temperature ≥22° C. and ≤30° C.

13. The process according to claim 1 , wherein a concentration of the carbon source in said second stage is <4 g/kg of medium.

14. The process according to claim 8 , wherein the carbon source is glucose.

15. The process according to claim 8 , wherein during an earlier stage of fermentation, the carbon source is being added, wherein the earlier stage is prior to the second stage.

16. The process according to claim 15 , wherein during said earlier stage the carbon source is in excess.

17. The process according to claim 8 , wherein the solvent is hexane.

18. The process according to claim 8 , wherein cells from the microorganism are pasteurized after fermentation is finished.

Assignees

• Dsm Ip Assets Bv

Inventors

• Streekstra Hugo
• Brocken Petrus Joseph Maria

Classifications

• A61K2800/85

  Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine · CPC title

• A61K8/67

  Vitamins · CPC title

• C12N1/005

  after treatment of microbial biomass not covered by C12N1/02 - C12N1/08 · CPC title

• A61K8/925

  of animal origin · CPC title

• A61Q19/00

  Preparations for care of the skin · CPC title