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]
- ↑ 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