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Main sol. 122

Internal Solution-ID: S220

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.3 9.838
MgCl2 x 6 H2O 2.60 g 2.6 12.789
KH2PO4 1.43 g 1.43 10.508
K2HPO4 5.50 g 5.5 31.577
CaCl2 x 2 H2O 0.13 g 0.13 0.884
Na2-ß-glycerophosphate x 5 H2O
(MERCK 35675)
6.00 g 6 19.601
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4)
1.10 ml 0.001 0.004
L-Glutathione
(reduced)
0.25 g 0.25 0.813
Yeast extract 4.50 g 4.5 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
Cellobiose 5.00 g 5 14.607
Cellulose
(Avicel or MN 301)
optional
10.00 g 10 -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except cellobiose, sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then adjust pH to 7.0 - 7.2, distribute under same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Cellobiose is added to the sterile medium from an anoxic 10% (w/v) stock solution prepared under 100% N2 gas and sterilized by filtration. Some strains can be adapted to cellulose as substrate using 10.00 g/l cellulose (Avicel or MN 301, MACHEREY-NAGEL).
2 Note: A white precipitate forms after mixing the ingredients of this medium, but this has no negative effect on growth.
122 ACETIVIBRIO MEDIUM