Methods and compositions for preserving bacteria
The disclosure provides methods and compositions for the preservation of bacteria.
1 . A composition comprising one or more bacterial strains belonging to the class Clostridia, a disaccharide other than lactose, an organic nitrogen source, an antioxidant, sodium metabisulfite, and a buffer at a pH of 6.7 to 7.3.
2 . The composition of claim 1 , wherein the disaccharide is sucrose.
3 . The composition of claim 2 , wherein the sucrose is at a concentration of 7.0% to 8.0%.
4 . The composition of claim 1 , wherein the disaccharide is trehalose.
5 . The composition of claim 4 , wherein the trehalose is at a concentration of 7.0% to 8.0%.
6 . The composition of claim 1 , wherein the sodium metabisulfite is at a concentration of 0.05%.
7 . The composition of claim 1 , wherein the buffer is at a pH of 7.0.
8 . The composition of claim 1 , wherein the composition comprises:
i) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 1;
(ii) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 2;
(iii) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 3;
(iv) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 4;
(v) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 5;
(vi) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 6;
(vii) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 7; and/or
(viii) a bacterial strain comprising a 16S rDNA sequence having at least 97% sequence identity to a nucleic acid sequence of SEQ ID NO: 8.
9 . The composition of claim 1 , wherein the organic nitrogen source is yeast extract, Luria-Bertani broth, or plant peptone.
10 . The composition of claim 1 , wherein the organic nitrogen source is yeast extract.
11 . The composition of claim 10 , wherein the yeast extract is at a concentration of 0.5% to 2.0%.
12 . The composition of claim 1 , wherein the antioxidant is inulin, riboflavin, or cysteine.
13 . The composition of claim 1 , wherein the antioxidant is cysteine.
14 . The composition of claim 13 , wherein the cysteine is at a concentration of 0.01% to 0.5%.
15 . The composition of claim 1 , wherein the buffer is a histidine buffer or a Tris buffer.
16 . The composition of claim 1 , wherein the composition comprises bacteria belonging to Clostridium clusters IV, XIVa, XVI, XVII, and/or XVIII.
17 . The composition of claim 1 , wherein the composition comprises one or more bacterial strains belonging to species selected from the group consisting of Clostridium bolteae, Anaerotruncus colihominis, Sellimonas intestinalis, Clostridium symbiosum, Blautia producta, Dorea longicatena, Clostridium innocuum , and Flavonifractor plautii.
18 . The composition of claim 1 , wherein the organic nitrogen source comprises a peptone.
19 . The composition of claim 18 , wherein the peptone is not derived from animal blood.
20 . A method for preserving bacteria of the class Clostridia, the method comprising
lyophilizing the composition of claim 1 .
21 . The method of claim 20 , wherein the bacterial strains belonging to the class Clostridia belong to Clostridium clusters IV, XIVa, XVI, XVII, and/or XVIII.