Method of control of Haematococcus spp, growth process

• US 5,882,849 A
• Filed: 07/01/1996
• Issued: 03/16/1999
• Est. Priority Date: 10/10/1989
• Status: Expired due to Fees

• Current Assignee: Aquasearch, Inc. (Mera Pharmaceuticals Incorporated)
• Original Assignee: Aquasearch, Inc. (Mera Pharmaceuticals Incorporated)
• Inventors: Redalje, Donald, Niiler, Pearn P., Huntley, Mark E., Leonard, Alexander B. P.
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US08/673,063
• Time in Patent Office: 988 Days
• Field of Search: 435/3, 435/41, 435/132, 435/166, 435/257.1, 435/257.3, 435/67, 435/257.6, 435/259, 435/292.1, 435/946, 47/1.4
• US Class Current: 435/3
• CPC Class Codes: C12M 21/02 Photobioreactors culturing algae A01G33/00, A01H4/001, C12N1/12
• CPC Class Codes: C12M 23/36 Means for collection or storage of gas; Gas holders
• CPC Class Codes: C12M 41/00 Means for regulation, monitoring, measurement or control, e.g. flow regulation controlling or regulating chemical, physical or physicochemical processes B01J19/0006; heating or cooling apparatus for laboratory use B01L7/00; electro optical investigation of individual particles, flow cytometers G01N15/14; automatic analysis G01N35/00; controlling or regulating in general G06N
• CPC Class Codes: C12M 41/06 of illumination
• CPC Class Codes: C12M 41/12 of temperature controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00
• CPC Class Codes: C12M 41/26 of pH
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• 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 control method for operating aqueous Haematococcus spp., such as H. pluvialis, microorganism growth processes is disclosed which can maintain viable growth conditions of this microorganism which have heretofore not been easily reproduced in commercially valuable quantities. The primary control parameters are the degree of turbulence in the aqueous growth medium and the scale of the apparatus relative to the scale of the turbulent eddies in vessels which are partially filled with the aqueous medium directly affect conditions which are required for optimum growth: light exposure, nutrient supply, sedimentation rate, bulk temperature, gas exchange rate and cell integrity. These control elements can be cast in terms of the Reynolds number (N_re), pH, temperature, amount of impinging light, and NO₂ concentration, depending upon the operative chlorophyll growth stages of Haematococcus spp. and its photoadaptive stages of producing astaxanthin.

Claims

1. A method for the control of aqueous Haematococcus and astaxanthin synthesis growth process which comprises:

distributing a photosynthetic Haematococcus biomass in an aqueous medium within a reaction chamber, said reaction chamber being transparent to visible light, and said aqueous medium containing said suspension of Haematococcus biomass occupying less than the total volume of said chamber;

maintaining said biomass within said reaction chamber for a 1-5 day period of time during which microorganisms comprising said biomass reproduce, increase in number, and retain their astaxanthin synthesis potential;

providing nutrients to and removing evolved gases from said biomass during said period of time to support said reproduction;

flowing quantities of said aqueous medium at a rate of 2-100 cm/sec through said reaction chamber throughout said time period;

subjecting said reaction chamber to an amount of visible light in the range 50-1000 μ

Em^-2 sec^-1 for at least a portion of said period of time with said visible light passing into said reaction chamber and being used by said microorganisms to enable photosynthesis;

maintaining a turbulent flow regime with Reynolds Number in the range of 2000 to 50,000 throughout said aqueous medium within said reaction chamber for said period of time;

maintaining a temperature in said chamber in the range of 16°

-27°

C. over the said time;

maintaining a pH in the range of 7.2 to 7.8 in said chamber over said period of time;

maintaining an NO₃^- control in the range of 4 to 12 mmoles/L in said chamber over said period of time;

at the end of said period of time harvesting the grown biomass from said reaction chamber and passing it to a ripening/maturation vessel;

maintaining a turbulent flow regime with a Reynolds Number in the range of 20,000 to 70,000 within said ripening/maturation vessel; and

maintaining a range of ambient light of 1500 to 2600 μ

Em^-2 sec^-1 in said vessel;

such that in the turbulent flow regimes effective reaction conditions are maintained for said reproduction of said Haematococcus microorganisms until said time period has been sufficient to produce a desired increase in Haematococcus and subsequently in production of astaxanthin by said Haematococcus.

2. A method as in claim 1 wherein each said turbulent flow regime causes movement of said Haematococcus such that exposure of individual microorganisms to said visible light is intermittent.

3. A method as in claim 2 wherein said intermittent exposure of visible light to Haematococcus creates a flashing light effect on photosynthesis produced in said microorganisms.

4. A method as in claim 3 wherein said Haematococcus microorganisms are exposed to said visible light for a cumulative time of approximately ten times longer than the cumulative time of non-exposure.

5. A method as in claim 1 wherein said aqueous medium containing said biomass occupies approximately 30%-90% of the total volume of said reaction chamber.

6. A method as in claim 1 wherein said Haematococcus microorganism comprises Haematococcus spp.

7. A method as in claim 6 wherein said Haematococcus spp. comprises H. pluvialis.

8. A method as in claim 1 wherein said Reynolds Number in said turbulent flow regime in said reaction chamber in the range of 5000-40,000.

9. A method as in claim 8 wherein said Reynolds Number in said turbulent flow regime in said reaction chamber in in the range of 10,000-30,000.

10. A method as in claim 1 wherein said amount of visible light to which said reaction chamber is exposed in in the range of 100-800 μ

Em^-2 sec^-1.

11. A method as in claim 10 wherein said amount of visible light to which said reaction chamber is exposed in in the range of 130-500 μ

Em^-2 sec^-1.

12. A method as in claim 1 further comprising passing said Haematococcus cells from said ripening/maturation vessel to a cell rupturing step wherein said cells are ruptured to free astaxanthin.

13. A method as in claim 12 wherein said rupturing comprises subjecting said cells to milling or ultrasonification.