CaGM

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Clostridium autoethanogenum Growth Medium

Composition (g/L)
NH4Cl 0.25
KCl 0.1
KH2PO4 0.2
MgSO4·7H2O 0.2
CaCl2·2H2O 0.02
Yeast Extract 1
2-g/liter resazurin 0.5 (ml)
2-(N-morpholino)ethanesulfonicacid(MES) 20
Fe(SO4)2·7H2O 0.05
Sodium Acetate · 3H2O 0.25
nitrilotriacetic acid (NTA) 0.05
Fructose (only for heterotrophic growth) 10
Trace Element (TSE) solution 10 (ml)
Wolfe's Vitamin Solution 10 (ml)
TSE Solution Composition (per liter)
nitrilotriaceticacid 2g
MnSO4·H2O 1g
Fe(SO4)2(NH4)2 · 6H2O 0.8g
CoCl2 · 6H2O 0.2g
ZnSO4·7H2O 0.2mg
CuCl2 ·2H2O 0.02g
NaMoO4·2H2O 0.02g
Na2SeO3 0.02
NiCl2 ·6H2O 0.02g
Na2WO4 ·2H2O 0.02g
Vitamin Solution Composition (per liter)
Biotin 2 mg
Folic Acid 2 mg
Pyridoxine hydrochloride 10 mg
Thiamine HCl 5 mg
Riboflavin 5 mg
Nicotinic Acid 5 mg
Calcium pantothenate 5 mg
Vitamin B12 0.1 mg
p-aminobenzoic acid 5 mg
thiotic acid 5 mg
Reducing Agent 1 (per 100 ml H2O)
cysteine HCl 4 g
Reducing Agent 2 (per 100 ml H2O)
NTA 7.64 g
Na2CO3 5.33 g
TiCl3 8.5 ml


Medium is prepared anaerobically and pH adjusted to 5.8 before sterilization. Prior to inoculation, 100 ml of CaGM is reduced with 1 ml of reducing agent 1 and 1 ml of reducing agent 2. [1]

  1. Liew FHenstra AM, Winzer K, Köpke M, Simpson SD, Minton NP.2016.Insights into CO2 Fixation Pathway of Clostridium autoethanogenum by Targeted Mutagenesis. mBio7:10.1128/mbio.00427-16.https://doi.org/10.1128/mbio.00427-16