|
KP buffer* |
5 |
ml |
|
MgCl2·6H2O |
3.05 |
g |
|
CaCl2·2H2O |
0.15 |
g |
|
(NH4)2SO4 |
0.66 |
g |
|
NaCl |
30 |
g |
|
Na2S2O3·5H2O |
5 |
g |
|
NaNO3 |
0.85 |
g |
|
Trace elements solution** |
2 |
ml |
|
Vitamin solution*** |
2 |
ml |
|
NaHCO3 |
2.52 |
g |
|
Distilled water |
1 |
L |
|
pH unajusted |
|
|
|
|
Mix ingredients except KP buffer and vitamin solution. Dispense the medium into suitable culture vessels under a stream of N2/CO2/O2 (75/20/5) and seal with butyl rubber stoppers. Separately autoclave KP buffer under N2 atmosphere. Sterilize vitamin solution by filtration. Prior to inoculation, aseptically add KP buffer and vitamin solution, and pressurize the inoculated vessels to 150 kPa with N2/CO2/O2 (75/20/5). |
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*KP buffer |
|
|
|
KH2PO4 |
119 |
g |
|
|
|
K2HPO4 |
21 |
g |
|
|
|
Distilled water |
1 |
L |
|
|
Autoclaved under N2. |
|
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|
**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 |
|
|
|
Distilled water |
1 |
L |
|
|
|
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add other minerals. Final pH 7.0. |
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***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 |