Methods for culturing and preserving
A method for culturing and preserving the probiotic gut microorganism Eubacterium hallii is disclosed. The method provides an E. hallii preparation that is suitable for administration or ingestion to humans and that provides high E. hallii biomass yields. Preferably, the medium used for culturing E. hallii to high biomass yields comprises food grade components only, is free of any animal sources, and/or is kosher. Further, including a method of treating disease using the E. hallii preparation.
1. A method of preparing an Eubacterium hallii preparation suitable for ingestion by a human, said method comprising the steps of:
a) providing a medium comprising:
1-100 g/kg of a sugar,
1-100 g/kg of a source of nitrogen and growth factors,
0.1-50 g/kg plant peptone,
acetate,
a buffer system at pH 6-8,
optionally, cysteine,
optionally, magnesium ions,
optionally, calcium ions,
optionally, potassium ions, and
optionally, vitamins;
b) inoculating said medium with E. hallii;
c) allowing said E. hallii to grow in said medium; and
d) harvesting said E. hallii preparation from said medium.
2. The method of claim 1 , which is carried out under anaerobic conditions.
3. The method of claim 1 , which is carried out at a temperature in the range of 30-42° C.
4. The method of claim 1 , wherein said E. hallii preparation is harvested from said medium using filtration.
5. The method of claim 4 , wherein the filtrate is further washed with phosphate buffered saline solution.
6. The method of claim 1 , which further comprises the step of lyophilizing said E. hallii to obtain a lyophilized E. hallii preparation.
7. The method of claim 6 , wherein said lyophilizing is carried out using a cryoprotectant selected from the group consisting of sucrose, maltose, maltodextrin, trehalose, mannitol, glycerol, DMSO, ethylene glycol, propylene glycol, 2-methyl-2,4-pentanediol, polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, polyglycerol, and any combination thereof.
8. The method according to claim 1 , wherein the medium comprises:
from 5 to 40 g/kg of sugar.
9. The method according to claim 1 , wherein the medium comprises:
from 5 to 40 g/kg of a source of nitrogen and growth factors.
10. The method according to claim 1 , wherein the sugar is glucose.
11. The method according to claim 1 , wherein the source of nitrogen and growth factors is yeast extract.
12. The method according to claim 1 , wherein the plant peptone is soy peptone.
13. The method according to claim 3 , wherein the temperature is in the range of 34-40° C.
14. The method according to claim 2 , carried out at a temperature in the range of 30-42° C.
15. The method according to claim 14 , wherein the E. hallii preparation is harvested from the medium utilizing filtration.
16. The method according to claim 15 , wherein the filtrate is further washed with phosphate buffered saline solution.
17. Eubacterium hallii preparation prepared by the method of claim 1 , wherein the E. hallii preparation comprises medium comprising:
sugar,
a source of nitrogen and growth factors,
plant peptone, and
acetate.
18. A medicament comprising the E. hallii preparation of claim 17 .
19. A method of treating insulin resistance and/or insulin resistance-related complications, metabolic syndrome, dyslipidemia, type 2 diabetes mellitus, non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatosis hepatitis (NASH), and insulin resistance in endocrine diseases, or for treating lactic-acidosis, short bowel syndrome or inflammatory bowel disease, or reducing the incidence or reducing the severity of colorectal cancer or colitis in a subject, the method comprising:
administering to the subject the E. hallii preparation of claim 17 .
20. A method to promote butyric acid formation in the intestine of a mammal, said method comprising the step of administering an E. hallii preparation according to claim 17 to a subject in need thereof.