881 .
KNH MEDIUM
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Equipment needed:
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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KH2PO4 | 0.136 | g | ||
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NH4Cl | 0.535 | g | ||
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MgCl2 x 6 H2O | 3.050 | g | ||
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CaCl2 x 2 H2O | 0.147 | g | ||
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NaHCO3 | 2.520 | g | ||
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NaCl | 20.450 | g | ||
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Na2SO4 | 2.840 | g | ||
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Sodium acetate | 0.656 | g | ||
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Trace element solution | 1.000 | ml | ||
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Se/W solution | 1.000 | ml | ||
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Trace vitamins | 1.000 | ml | ||
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Resazurin | 1.000 | mg | ||
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Sodium thioglycolate | 0.100 | g | ||
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Ascorbic acid | 0.100 | g | ||
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Distilled water | 1000.000 | ml | ||
| 1 Mix all ingredients except sodium thioglycolate and ascorbic acid, and expel dissolving O2 from the solution with deoxygenated N2-CO2 (80:20, v/v). Dispense the solution into vials where deoxygenated H2-CO2 (80:20, v/v) is sparged. Cap with butyl rubber stoppers. Autoclave, then add reducing agent solution (sodium thioglycolate and ascorbic acid, 1% solution each) sterilized with 0.2 �m-filter (0.1 ml in 10 ml medium) and inoculate fully grown culture at 5-10% of final volume. Fill with H2-CO2 (80:20, v/v) gas at 1.5 atm. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Nitrilotriacetic acid | 12.800 | g | ||
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FeCl3 x 6 H2O | 1.350 | g | ||
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MnCl2 x 4 H2O | 0.100 | g | ||
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CoCl2 x 6 H2O | 0.024 | g | ||
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CaCl2 x 2 H2O | 0.100 | g | ||
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ZnCl2 | 0.100 | g | ||
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CuCl2 x 2 H2O | 0.025 | g | ||
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H3BO3 | 0.010 | g | ||
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NaCl | 1.000 | g | ||
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NiCl2 x 6 H2O | 0.120 | g | ||
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Na2SeO3 x 5 H2O | 0.026 | g | ||
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Distilled water | 1000.000 | ml | ||
| 1 Dissolve nitrilotriacetic acid first in 200 ml of distilled water, and adjust pH to 6.5 with KOH. Then add the remaining salts and adjust volume to 1.0 L. Readjust pH to 7.0 with KOH. | |||||
| 2 Comment: For strain JCM 10134, incubate with a 10-20% inoculum. | |||||
Se/W solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2SeO3 x 5 H2O | 4 | mg |
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Na2WO4 x 2 H2O | 4 | mg |
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Distilled water | 1000 | ml |
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
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Biotin | 2.0 | mg |
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Folic acid | 2.0 | mg |
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Pyridoxine hydrochloride | 10.0 | mg |
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Thiamine HCl | 5.0 | mg |
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Riboflavin | 5.0 | mg |
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Nicotinic acid | 5.0 | mg |
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Calcium pantothenate | 5.0 | mg |
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Vitamin B12 | 0.1 | mg |
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p-Aminobenzoic acid | 5.0 | mg |
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Lipoic acid | 5.0 | mg |
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Distilled water | 1000.0 | ml |