Procedure for large-scale production of astaxanthin from haematococcus

• US 6,022,701 A
• Filed: 07/30/1998
• Issued: 02/08/2000
• Est. Priority Date: 02/01/1996
• Status: Expired due to Term

• Current Assignee: Ben Gurion University of The Negev
• Original Assignee: Ben Gurion University of The Negev
• Inventors: Vonshak, Avigad, Richmond, Amos, Cohen, Zvi, Boussiba, Sammy
• Primary Examiner(s): Lilling, Herbert J.
• Application Number:
  US09/117,497
• Time in Patent Office: 558 Days
• Field of Search: 435/67, 435/257.1
• US Class Current: 435/67
• CPC Class Codes: C12P 23/00 Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes containing heterorings C12P17/00

Abstract

A process for cultivating Haematococcus for the large scale production of astaxanthin-enriched Haematococcus cells comprises: (a) cultivating said Haematococcus cells under conditions suitable for optimal vegetative growth of said cells, wherein said conditions comprise growing the cells under a light intensity in the range of about 30-140 μmol photons.m^-2.S^-1 and at a temperature of between about 15-28° C.; and (b) collecting the cells grown according to (a) and cultivating them further under conditions suitable for optimal induction and accumulation of astaxanthin in said cells, wherein said conditions comprise inoculating said cells of (a) into a growth solution containing essentially a carbon source and growing said cells at a temperature of below 35° C.

Claims

1. A process for cultivating Haematococcus cells for the large scale production of astaxanthin-enriched Haematococcus cells comprising:

(a) cultivating said Haematococcus cells under conditions suitable for optimal vegetative growth of said cells, wherein said conditions comprise growing the cells under a light intensity in the range of about 30-140 μ

mol photons.m^-2·

S^-1 and at a temperature of between about 15-28°

C.; and

(b) collecting the cells grown according to (a) and cultivating them further under conditions suitable for optimal induction and accumulation of astaxanthin in said cells, wherein said conditions comprise inoculating said cells of (a) into a growth solution consisting essentially of tap water to which is added CO₂ as the carbon source for the cells, and growing said cells at a temperature of below 35°

C.

2. A process according to claim 1, wherein said optimal vegetative growth conditions of (a) are by growing said cells in a photobioreactor to facilitate the controlled environmental conditions of said light intensity and temperature.

3. A process according to claim 2, wherein said photobioreactor is a vertical photobioreactor.

4. A process according to claim 2, wherein said photobioreactor is a tubular panel photobioreactor.

5. A process according to claim 1, wherein the growth conditions of (a) further comprise growing said cells in Modified BG-11 medium and agitation of the cells by a flow of a mixture of CO₂ in air through the culture.

6. A process according to claim 1, wherein the initial concentration of cells used for the cultivation in (a) is about 0.1-0.3×

10⁶ cells/ml.

7. A process according to claim 1, wherein the cultivating step (a) comprises the conditions of growing the cells under a light intensity in the range of about 70-90 μ

mol photons.m^-2.s^-1 and at a temperature of between about 22-25°

C.

8. A process according to claim 1, wherein said optimal conditions for the induction and accumulation of astaxanthin in the cells of (b) are by growing said cells in a photobioreactor to facilitate the effective maintenance of the temperature at below 35°

C.

9. A process according to claim 8, wherein said photobioreactor is a tubular bioreactor.

10. A process according to claim 8, wherein said cultivation of (b) is carried out outdoors and the light source for the cells is sunlight.

11. A process according to claims 8, wherein said cultivation of (b) is carried out under conditions of temperature control at below 32°

C.

12. A process for the large-scale production of astaxanthin-enriched Haematococcus cells comprising cultivating Haematococcus cells according to claim 1 and further comprising the steps:

(c) harvesting the cells cultivated in (b) by collecting said cells, concentrating the collected cells, drying the concentrated cells and freezing the dried cells; and

(d) processing the frozen dried cells of (c) by grinding them under cryogenic conditions in the presence of a suitable antioxidant to obtain an astaxanthin-enriched product.

13. A process according to claim 12, wherein the collection and concentration of said cells is by sedimentation followed by centrifugation or filtration under vacuum, and wherein the drying of the said cells is by lyophilization, or by spray drying.

14. A process according to claim 12, wherein the grinding of said dry cells is by way of an impact mill.

15. A process according to claim 12, wherein the antioxidant is selected from the group consisting of ethoxyquin, butylated hydrox yanisole, butylated hydroxytoluene (BHT), tocopherols, di-tert-butyl-paracresol and propyl gallate.

16. A process according to claim 1, wherein the Haematococcus cells are of the strain Haematococcus pluvialis.

17. A process according claim 12, wherein the astaxanthin-enriched cells are Haematococcus pluvialis cells.