Multi-stage process for production of immune modulator
Immune function of an animal can be modulated by administration of a composition that includes algae meal or beta glucan. The algae meal can be made by growing Euglena using particular methods and conditions, including certain continuous, semi-continuous, fed-batch, and repeat batch methods in sterile fermenters. Euglena provides a form of beta glucan that is different from other organisms, where the beta glucan is predominantly unbranched beta-1,3-glucan. Use of algae meal and beta glucan produced by the disclosed processes can improve the wellbeing of an animal or human, and may augment or even replace the use of antibiotics in certain circumstances.
1. A method for growing Euglena comprising:
(a) growing Euglena heterotrophically in a growth media in a vessel to form a culture, wherein the Euglena are grown to a concentration of at least 50 grams dry weight per liter and the Euglena have greater than 30% by weight beta glucan and less than 70% by weight beta glucan;
(b) removing a portion of the culture from the vessel to form a removed portion; and
(c) replenishing the remaining culture in the vessel with fresh growth media to form a replenished culture;
wherein the productivity of Euglena grown by steps (a), (b), and (c) is at least 20 grams dry weight per liter of culture maintained in the vessel per day, and
wherein a carbon source in the growth media in steps (a) and (c) is less than 30 grams per liter.
2. The method of claim 1 , wherein the Euglena in the culture of step (a) have greater than 40% by weight beta glucan and less than 60% by weight beta glucan.
3. The method of claim 1 , further comprising growing Euglena in the replenished culture of step (c) to a concentration of at least 50 grams dry weight per liter.
4. The method of claim 3 , further comprising:
(d) removing a portion of the replenished culture from the vessel to form a removed replenished portion.
5. The method of claim 4 , further comprising:
(e) replenishing the remaining culture in the vessel with fresh growth media.
6. The method of claim 4 , further comprising combining the removed portions of steps (b) and (d).
7. The method of claim 6 , further comprising one of:
separating the combined removed portions into a solid portion comprising the Euglena and a liquid portion, and drying the solid portion to form an algae biomass; and
extracting the combined removed portions to isolate paramylon from the Euglena.
8. The method of claim 1 , further comprising repeating steps (a), (b), and (c) a plurality of times.
9. The method of claim 1 , wherein steps (b) and (c) are performed simultaneously.
10. The method of claim 1 , wherein steps (b) and (c) are performed sequentially.
11. The method of claim 1 , wherein the vessel comprises a volume of at least 100 liters.
12. The method of claim 1 , wherein the growth media in steps (a) and (c) is sterilized.
13. The method of claim 1 , wherein step (a) uses an air lift or bubble column reactor.
14. The method of claim 1 , wherein step (a) uses an agitator.
15. The method of claim 1 , wherein the doubling rate of Euglena in step (a) is between 10 and 30 hours.
16. The method of claim 1 , wherein the growth media in steps (a) and (c) comprises ammonium hydroxide as the primary nitrogen source.
17. The method of claim 1 , wherein the growth media in steps (a) and (c) comprises a citrate-citric acid buffer.
18. The method of claim 1 , wherein the growth media in steps (a) and (c) comprises ethanol as a carbon source.
19. The method of claim 1 , wherein the removed portion of step (b) comprises less than 2 grams of a carbon source for every 1 gram of Euglena.
20. The method of claim 1 , further comprising separating the removed portion of step (b) into a solid portion comprising the Euglena and a liquid portion.
21. The method of claim 20 , further comprising drying the solid portion to form an algae biomass.
22. The method of claim 21 , further comprising reducing the average particle size of the algae biomass to 1000 microns or less.
23. The method of claim 21 , further comprising reducing the average particle size of the algae biomass to 500 microns or less.
24. The method of claim 1 , further comprising extracting the removed portion of step (b) to isolate paramylon from the Euglena.
25. The method of claim 1 , wherein the vessel is a bioreactor.
26. The method of claim 1 , wherein the volume of fresh growth media in step (c) is equal to the volume of the removed portion in step (b).
27. The method of claim 1 , wherein the vessel comprises a volume of at least 5000 liters.
28. The method of claim 1 , wherein step (a) does not use an agitator blade.
29. The method of claim 1 , wherein steps (a), (b), and (c) are repeated for up to 25 days.