| |
KH2PO4 |
0.75 |
g |
| |
K2HPO4 |
0.75 |
g |
| |
NH4Cl |
1 |
g |
| |
MgCl2·6H2O |
0.36 |
g |
| |
Bacto Yeast Extract (Difco) |
0.1 |
g |
| |
Polypeptone |
0.1 |
g |
| |
Trace elements solution* |
10 |
ml |
| |
Vitamin solution** |
10 |
ml |
| |
Resazurin |
1 |
mg |
| |
NaHCO3 |
4 |
g |
| |
Cysteine-HCl |
0.5 |
g |
| |
Na2S·9H2O |
0.5 |
g |
| |
Distilled water |
1 |
L |
| |
| |
|
Mix ingredients except for vitamin solution, cysteine-HCl and Na2S·9H2O, and autoclave under H2/CO2 (80/20) condition. Separately autoclave cysteine-HCl and Na2S·9H2O as a 5% solution under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin solution, and cysteine-HCl and Na2S·9H2O solutions, and pressurize inoculated bottles to 150 kPa with H2/CO2. |
| |
|
|
|
|
|
|
| |
|
*Trace elements solution |
| |
|
|
Nitrilotriacetic acid (NTA) |
4.1 |
g |
| |
|
|
FeCl3·6H2O |
0.36 |
g |
| |
|
|
MnCl2·4H2O |
0.09 |
g |
| |
|
|
CoCl2·6H2O |
0.15 |
g |
| |
|
|
CaCl2·2H2O |
0.018 |
g |
| |
|
|
ZnCl2 |
0.09 |
g |
| |
|
|
H3BO3 |
0.017 |
g |
| |
|
|
Na2MoO4·2H2O |
0.009 |
g |
| |
|
|
NiCl2·6H2O |
0.018 |
g |
| |
|
|
Distilled water |
1 |
L |
| |
| |
|
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0. |
| |
|
|
|
|
|
|
| |
|
**Vitamin solution |
| |
|
|
Biotin |
2 |
mg |
| |
|
|
Folic acid |
2 |
mg |
| |
|
|
Pyridoxine-HCl |
10 |
mg |
| |
|
|
Thiamine-HCl |
5 |
mg |
| |
|
|
Riboflavin |
5 |
mg |
| |
|
|
Nicotinic acid |
5 |
mg |
| |
|
|
Ca-pantothenate |
5 |
mg |
| |
|
|
p-Aminobenzoic acid |
1 |
mg |
| |
|
|
Vitamin B12 |
0.01 |
mg |
| |
|
|
Distilled water |
1 |
L |
|