Methods and compositions for preserving bacteria
The disclosure provides methods and compositions for the preservation of bacteria.
1. A method for preserving bacteria, the method comprising:
(i) adding bacteria to a composition comprising a disaccharide, yeast extract, cysteine, sodium metabisulfite, and a histidine buffer; and
(ii) subjecting the composition comprising the bacteria to a lyophilization cycle.
2. The method of claim 1 , wherein the lyophilization cycle comprises one or more steps of a temperature ramp rate between 0.5° C./min to 3° C./min.
3. The method of claim 2 , wherein the temperature ramp rate is 1° C./min.
4. The method of claim 2 , wherein the temperature ramp rate is 2.5° C./min.
5. The method of claim 1 , wherein the method further comprises measuring the number of colony forming units after subjecting the composition comprising the bacteria to the lyophilization cycle.
6. The method of claim 5 , wherein the method further comprises measuring the number of colony forming units prior to subjecting the composition comprising the bacteria to the lyophilization cycle.
7. The method of claim 6 , wherein the method further comprises:
comparing the number of colony forming units prior to subjecting the composition comprising the bacteria to the lyophilization cycle and the number of colony forming units after subjecting the composition comprising the bacteria to the lyophilization cycle; and
determining a percentage of recovered colony forming units of the bacteria after the lyophilization cycle.
8. The method of claim 1 , wherein the composition maintains viability of one or more bacterial strains upon lyophilization.
9. The method of claim 1 , wherein the method allows for recovery of at least 10% of colony forming units of the bacteria over a period of time.
10. The method of claim 9 , wherein the period of time is at least 2 weeks.
11. The method of claim 1 , wherein the method allows for recovery of at least 10% of colony forming units of the bacteria after a freeze-thaw cycle.
12. The method of claim 1 , wherein the method allows for recovery of at least 10% of colony forming units of the bacteria after the lyophilization cycle.
13. The method of claim 1 , wherein the bacteria comprise one or more bacterial strains belonging to the class Clostridia.
14. The method of claim 1 , wherein the bacteria comprise one or more bacterial strains belonging to the family Clostridiaceae.
15. The method of claim 1 , wherein the bacteria comprise one or more bacterial strains belonging to Clostridium clusters IV and/or XIVa.
16. The method of claim 1 , wherein the bacteria comprise one or more bacterial strains belonging to species selected from the group consisting of Clostridium bolteae, Anaerotruncus colihominis, Ruminococcus torques, Clostridium symbiosum, Blautia producta, Dorea longicatena, Erysipelotrichaceae bacterium and Subdolinogranulum spp.
17. The method of claim 1 , wherein the disaccharide is trehalose.
18. The method of claim 17 , wherein the composition comprises about 7.5% trehalose, about 1% yeast extract, about 0.05% cysteine, about 0.05% sodium metabisulfite, and about 20 mM histidine buffer.
19. The method of claim 1 , wherein the disaccharide is sucrose.
20. The method of claim 19 , wherein the composition comprises 7.0% sucrose, 1% yeast extract, 0.05% cysteine, 0.05% sodium metabisulfite, and 20 mM histidine buffer.