Method of mixotrophic culturing of microalgae in a flexible bioreactor

Patent metadata

Field	Value
Publication number	US-10047337-B2
Application number	US-201615142913-A
Country	US
Kind code	B2
Filing date	Apr 29, 2016
Priority date	Mar 31, 2015
Publication date	Aug 14, 2018
Grant date	Aug 14, 2018

Abstract

Some embodiments include a method of culturing one or more microalgae. The method can include: inoculating a bioreactor with the one or more microalgae and a fluidic support medium, the bioreactor having one or more bioreactor walls at least partially enclosing a bioreactor cavity, being configured to be at least one of folded up or rolled up, and being sterile when the bioreactor is inoculated with the one or more microalgae, the one or more bioreactor walls comprising at least one bioreactor wall material, and the at least one bioreactor wall material being flexible and at least partially transparent; and vitally supporting the one or more microalgae. Other embodiments of related systems and methods are also disclosed.

First claim

What is claimed is:

1. A method of culturing microalgae, the method comprising:
inoculating a bioreactor with microalgae and a fluidic support medium, the bioreactor comprising one or more bioreactor walls at least partially enclosing a bioreactor cavity, being configured to be at least one of folded up or rolled up, and being sterile when the bioreactor is inoculated with the microalgae, the one or more bioreactor walls comprising at least one bioreactor wall material, the at least one bioreactor wall material being flexible and at least partially transparent;
and
vitally supporting the microalgae, wherein the microalgae are taxonomically classified in taxonomic family Chlorellaceae;
wherein:
vitally supporting the microalgae comprises:
supplying the microalgae with an organic carbon material mixed with a nitrate comprising acetic acid in an amount up to approximately 40% and sodium nitrate in an amount up to approximately 4%;
supplying the microalgae with a quantity of light based on the culture density;
supplying one or more nutritional media comprising magnesium sulfate heptahydrate in an amount up to approximately 150 milligrams per liter, trace metals in an amount up to approximately 0.5 milliliters per liter, and phosphate dibasic in an amount up to approximately 200 milligrams per liter, to the microalgae;
operating the bioreactor in substantially axenic conditions; and
mixing the microalgae within the fluidic support medium; and

mixing the microalgae within the fluidic support medium comprises:
injecting gas into the fluidic support medium with only one gas delivery device; and
the gas comprises gas bubbles having a diameter greater than or equal to approximately 40 micrometers and less than or equal to approximately 2 millimeters.

2. The method of claim 1 wherein:
injecting the gas into the fluidic support medium with only one gas delivery device comprises:
injecting the gas into the fluidic support medium at a volumetric flow rate of greater than or equal to approximately 10 liters per minute and less than or equal to approximately 60 liters per minute.

3. The method of claim 2 wherein:
the bioreactor has a length dimension of greater than or equal to approximately 182 centimeters and less than or equal to approximately 244 centimeters; and
mixing the microalgae within the fluidic support medium further comprises:
supplying one or more surfactants into the fluidic support medium.

4. The method of claim 1 further comprising:
partially harvesting the microalgae from the bioreactor at intervals of 7-12 days.

5. The method of claim 1 further comprising:
partially harvesting the microalgae from the bioreactor at intervals of 10-12 days.

6. The method of claim 1 further comprising:
partially harvesting the microalgae from the bioreactor when the culture density comprises at least approximately 2 grams per liters.

7. The method of claim 1 further comprising:
partially harvesting the one or more microalgae from the bioreactor when the culture density is greater than or equal to approximately 2 grams per liter and less than or equal to approximately 5 grams per liter.

8. The method of claim 1 further comprising:
partially harvesting the one or more microalgae from the bioreactor when the culture density is greater than or equal to approximately 7 grams per liter and less than or equal to approximately 12 grams per liter.

9. The method of claim 1 further comprising:
partially harvesting the one or more microalgae from the bioreactor when the culture density is greater than or equal to approximately 7 grams per liter and less than or equal to approximately 10 grams per liter.

10. The method of claim 1 further comprising:
partially harvesting the microalgae from the bioreactor when the culture density is greater than or equal to approximately 20 grams per liter and less than or equal to approximately 30 grams per liter.

11. The method of claim 1 wherein:
supplying the microalgae with the quantity of light based on the culture density comprises:
doubling the quantity of light supplied to the microalgae when the culture density exceeds approximately 0.5 grams per liter.

12. The method of claim 1 wherein:
the one or more microalgae are taxonomically classified in taxonomic family Haematococcaceae; and
vitally supporting the one or more microalgae further comprises at least one of:
culturing the one or more microalgae such that the culture density is greater than or equal to approximately 12 grams per liter; or
culturing the one or more microalgae such that the microalgae grows at an average maximum production rate greater than or equal to approximately 2.5 grams per liter day.

13. The method of claim 1 wherein:
vitally supporting the microalgae further comprises at least one of:
culturing the microalgae such that the culture density is greater than or equal to approximately 36 grams per liter; or
culturing the microalgae such that the microalgae grows at an average maximum production rate greater than or equal to approximately 9 grams per liter day.

14. The method of claim 1 wherein:
the one or more microalgae are taxonomically classified in taxonomic family Chlamydomonadaceae; and
vitally supporting the one or more microalgae further comprises at least one of:
culturing the one or more microalgae such that the culture density is greater than or equal to approximately 7 grams per liter; or
culturing the one or more microalgae such that the microalgae grows at an average maximum production rate greater than or equal to approximately 3 grams per liter day.

15. A method of culturing microalgae, the method comprising:
inoculating a bioreactor with microalgae and a fluidic support medium, the bioreactor comprising one or more bioreactor walls at least partially enclosing a bioreactor cavity, being configured to be at least one of folded up or rolled up, and being sterile when the bioreactor is inoculated with the microalgae, the one or more bioreactor walls comprising at least one bioreactor wall material, the at least one bioreactor wall material being flexible and at least partially transparent;
and
vitally supporting the microalgae, wherein the microalgae are taxonomically classified in taxonomic family Chlorellaceae;

wherein:
vitally supporting the microalgae comprises:
supplying the microalgae with an organic carbon material mixed with a nitrate, trace metals, and a magnesium sulfate comprising acetic acid in an amount up to approximately 40%, sodium nitrate in an amount up to approximately 4%, trace metals in an amount up to approximately 3.34 milligrams per liter, and magnesium sulfate heptahydrate in an amount up to approximately 6.67 milligrams per liter;
supplying the microalgae with a quantity of light based on the culture density;
supplying one or more nutritional media comprising phosphate dibasic in an amount up to approximately 200 milligrams per liter, to the microalgae when a culture density of the microalgae reaches threshold culture densities of approximately 5 grams per liter and approximately 15 grams per liter;
operating the bioreactor in substantially axenic conditions; and
mixing the microalgae within the fluidic support medium; and mixing the microalgae within the fluidic support medium comprises: injecting gas into the fluidic support medium with only one gas delivery device; and the gas comprises gas bubbles having a diameter greater than or equal to approximately 40 micrometers and less than or equal to approximately 2 millimeters.

16. The method of claim 15 wherein:
the b

Assignees

• Heliae Dev Llc
• Heliae Dev Llc

Inventors

• Cizek Luke
• Olaizola Miquel
• Mccarty Mason
• Oelschlager Mason

Classifications

• C12M41/22

  in contact with the bioreactor walls · CPC title

• C12M27/04

  with introduction of gas through the stirrer or mixing element · CPC title

• C12M29/04

  Filters; Permeable or porous membranes or plates, e.g. dialysis · CPC title

• C12M37/00

  Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination (C12M23/38 takes precedence; filtration in general and filters per se B01D24/00-B01D41/00; autoclaves B01J3/04; treatment of microorganisms with electrical or wave energy C12N13/00) · CPC title

• C12M23/48

  Holding appliances; Racks; Supports (holding devices for laboratory apparatus B01L9/00) · CPC title