Bioreactor for the selection of microalgae

Patent metadata

Field	Value
Publication number	US-11427796-B2
Application number	US-201716335390-A
Country	US
Kind code	B2
Filing date	Sep 19, 2017
Priority date	Sep 21, 2016
Publication date	Aug 30, 2022
Grant date	Aug 30, 2022

Abstract

The invention relates to a bioreactor comprising a tank ( 100 ) capable of being operated for a working period, said tank ( 100 ) being intended to receive a culture medium comprising a cellular culture of photosynthetic microorganisms, a light source ( 200 ) arranged to emit incident light having a chosen incoming light intensity (Iin) in the direction of the tank, a temperature probe ( 400 ) for measuring the temperature of said culture medium in the tank, and a temperature regulator ( 500 ) capable of raising and lowering the temperature of said culture medium in the tank, and further comprising a control ( 700 ) of the temperature regulator arranged to adjust the temperature of the culture medium to a low setpoint value (VCB) during a first period, and to adjust the temperature of the culture medium to a high setpoint value (VCH) during a second period, the succession of said first and second periods making it possible to induce a cellular stress in at least some of said photosynthetic microorganisms during the working period.

First claim

The invention claimed is:

1. A bioreactor characterized in that it comprises a tank capable of being operated during a working period, and capable of repeating the working period once it has been completed, said tank being intended to receive a culture medium comprising a cell culture of photosynthetic microorganisms, a light source arranged to emit incident light having a chosen incoming light intensity (I in ) in the direction of the tank, a temperature probe for measuring the temperature of said culture medium in the tank, and a temperature regulator capable of increasing and decreasing the temperature of said culture medium in the tank, and in that the tank also comprises a control of the temperature regulator arranged to adjust the temperature of the culture medium to a low setpoint value (V LS ) during a first period, and to adjust the temperature of the culture medium to a high setpoint value (V HS ) during a second period, the succession of said first and second periods making it possible to induce a cellular stress in at least some of said photosynthetic microorganisms during the working period, and wherein the control is also arranged to receive data relating to the growth rate of said cell culture of photosynthetic microorganisms after each working period of a plurality of working periods, and wherein the control fixes the low setpoint value and said high setpoint value for a subsequent working period after said reception of the data relating to the growth rate of said cell culture of photosynthetic microorganisms from a prior working period, and wherein the control is arranged to determine said low setpoint value and said high setpoint value and said first and second periods so as to maintain a mean setpoint value (V M ) during each working period, the mean value being equal to the sum of the product of said low setpoint value and of said first period and of the product of said high setpoint value and of said second period, divided by the working period, and wherein the plurality of working periods includes more than two working periods so as to bring about a cellular degradation of at least one portion of said photosynthetic microorganisms and thus to select the photosynthetic microorganisms having valuable properties.

2. The bioreactor according to claim 1 , wherein said mean setpoint value is substantially equal to the optimal growth temperature (T opt ) of the cell culture of photosynthetic microorganisms.

3. The bioreactor according to claim 1 , wherein said chosen incoming light intensity (I in ) of the incident light is fixed during the working period.

4. The bioreactor according to claim 1 , wherein said chosen incoming light intensity (I in ) of the incident light is between 100 and 2000 μmol quanta m −2 s −1 .

5. The bioreactor according to claim 1 , also comprising a controller arranged to operate the tank at a chosen cell culture concentration (x i ), in the culture medium during said working period.

6. The bioreactor according to claim 1 , also comprising a light sensor facing the light source, the sensor being capable of measuring an outgoing light intensity (I out ) and of transmitting data relating to this intensity (I out ) to the controller in order to calculate the concentration of the cell culture and to operate the tank at said chosen cell culture concentration (x i ) in the culture medium during the working period.

7. The bioreactor according to claim 1 , also comprising a controller arranged to operate the tank at a cell culture concentration (x i ) substantially less than or equal to 1.0 g/l in the culture medium during said working period.

8. The bioreactor according to claim 1 , wherein said chosen incoming light intensity (I in ) of the incident light is equal to approximately 250 μmol quanta m −2 s −1 .

9. A method for selecting photosynthetic microorganisms, comprising the following steps:
1. Providing a bioreactor according to claim 1 ;
2. Filling the tank with a culture medium;
3. Inoculating the culture medium with a cell culture consisting of photosynthetic microorganisms;
4. Operating the tank during a working period and adjusting the incoming light intensity (I in ) to a chosen value;
5. Adjusting the temperature of the culture medium to a low setpoint value (V L s) during a first period, and adjusting the temperature of the culture medium to a high setpoint value (V HS ) during a second period, the succession of said first and second periods making it possible to induce a cellular stress in at least some of said photosynthetic microorganisms during the working period;
6. Harvesting the photosynthetic microorganisms,

and wherein the temperature adjustment step 5. is repeated more than two times so as to bring about a cellular degradation of at least one portion of said photosynthetic microorganisms and thus to select the photosynthetic microorganisms having valuable properties.

10. The method according to claim 9 , wherein said low setpoint value and said high setpoint value and said first and second periods are adjusted so as to maintain a mean setpoint value (V M ) during the working period, the mean value being equal to the sum of the product of said low setpoint value and of said first period and of the product of said high setpoint value and of said second period, divided by the working period.

11. The method according to claim 9 , wherein said mean setpoint value is substantially equal to the optimal growth temperature (T opt ) of the cell culture of photosynthetic microorganisms.

12. The method according to claim 9 , also comprising the following steps:
4a. Taking a portion of the culture medium comprising at least one portion of said photosynthetic microorganisms; and
4b. Compensating for said portion of the culture medium comprising at least one portion of said photosynthetic microorganisms taken, with fresh culture medium.

13. The method according to claim 9 , wherein the harvesting in step 6. is carried out when the culture medium comprises a cell culture of photosynthetic microorganisms consisting of more than 75% of photosynthetic microorganisms having valuable properties.

14. The method according to claim 13 , also comprising the following steps:
7. Inoculating a fresh culture medium with the photosynthetic microorganisms having valuable properties in a bioreactor; and
8. Operating the bioreactor of step 7. for a production of biomass consisting of said photosynthetic microorganisms having valuable properties.

15. The method according to claim 9 , wherein the harvesting in step 6. is carried out when the culture medium comprises a cell culture of photosynthetic microorganisms consisting of more than 90% of photosynthetic microorganisms having valuable properties.

16. The method according to claim 9 , wherein the harvesting in step 6. is carried out when the culture medium comprises a cell culture of photosynthetic microorganisms consisting of substantially 100% of photosynthetic microorganisms having valuable properties.

Assignees

• Inria Inst Nat Rech Informatique & Automatique
• Centre Nat Rech Scient
• Univ Nice Sophia Anitpolis

Inventors

• Bernard Olivier
• Bonnefond Hubert
• Sciandra Antoine
• Pruvost Éric
• Grimaud Ghjuvan

Classifications

• C12N1/12

  Unicellular algae; Culture media therefor (as new plants A01H13/00) · CPC title

• C12M41/48

  Automatic or computerized control (automatic analysis G01N35/00) · CPC title

• C12M41/06

  of illumination · CPC title

• C12M41/12

  of temperature (controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00) · CPC title

• C12M21/02 Primary

  Photobioreactors (culturing algae A01G33/00, A01H4/001, C12N1/12) · CPC title