|
NH4Cl |
0.5 |
g |
|
K2HPO4 |
0.4 |
g |
|
MgCl2·6H2O |
0.1 |
g |
|
CaCl2·2H2O |
0.1 |
g |
|
Sodium acetate |
3.3 |
g |
|
Coenzyme M |
0.14 |
g |
|
NaHCO3 |
1 |
g |
|
Vitamin solution* |
10 |
ml |
|
Trace element solution** |
10 |
ml |
|
Resazurin |
0.5 |
mg |
|
Na2S·9H2O |
0.25 |
g |
|
Distilled water |
1 |
L |
|
|
|
Mix ingredients except for CaCl2·2H2O, sodium acetate, coenzyme M, NaHCO3, vitamin solution and Na2S·9H2O, and autoclave under a N2/CO2 atmosphere (80/20). Concentrated CaCl2·2H2O, sodium acetate, coenzyme M and Na2S·9H2O solutions are sterilized separately under a N2 atmosphere in tightly closed vessels. Concentrated NaHCO3 solution and vitamin are filter-sterilized. Prior to inoculation, add CaCl2·2H2O, sodium acetate, coenzyme M, NaHCO3, vitamin and Na2S·9H2O solutions. Final pH is 6.5. |
|
|
|
|
|
|
|
|
|
*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 |
|
|
|
|
|
|
|
|
|
**Trace elements solution |
|
|
|
Nitrilotriacetic acid (NTA) |
12.8 |
g |
|
|
|
FeCl3·6H2O |
1.35 |
g |
|
|
|
MnCl2·4H2O |
0.1 |
g |
|
|
|
CoCl2·6H2O |
0.024 |
g |
|
|
|
CaCl2·2H2O |
0.1 |
g |
|
|
|
ZnCl2 |
0.1 |
g |
|
|
|
CuCl2·2H2O |
0.025 |
g |
|
|
|
H3BO3 |
0.01 |
g |
|
|
|
Na2MoO4·2H2O |
0.024 |
g |
|
|
|
NaCl |
1 |
g |
|
|
|
NiCl2·6H2O |
0.12 |
g |
|
|
|
Na2SeO4 |
0.004 |
g |
|
|
|
Na2WO4 |
0.004 |
g |
|
|
|
KAl(SO4)2·12H2O |
0.02 |
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. |
|