|
KH2PO4 |
0.14 |
g |
|
NH4Cl |
0.54 |
g |
|
MgCl2·6H2O |
0.2 |
g |
|
CaCl2·2H2O |
0.15 |
g |
|
Bacto Yeast Extract (Difco) |
0.1 |
g |
|
Sodium acetate |
1 |
g |
|
Trace elements solution* |
1 |
ml |
|
Vitamin solution** |
2 |
ml |
|
Resazurin |
1 |
mg |
|
NaHCO3 |
2.5 |
g |
|
Cysteine-HCl |
0.3 |
g |
|
Na2S·9H2O |
0.3 |
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 6% 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 |
|
|
|
Na2SeO4 |
0.004 |
g |
|
|
|
Na2WO4 |
0.004 |
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 |
|