81 .
MINERAL MEDIUM FOR CHEMOLITHOTROPHIC GROWTH (H-3)
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Selected strain: Herbaspirillum autotrophicum DSM 733
This strain uses the standard medium without modifications.
Growth conditions: 28 °C
Equipment needed:
Autoclave
Filter
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Solution A | 50 | ml | ||
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Solution B | 925 | ml | ||
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Solution C | 20 | ml | ||
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Standard vitamin solution | 5 | ml | ||
| 1 Solutions A, B, C are autoclaved separately for 15 min at 121°C, cooled down to 50°C and then mixed aseptically with 5.0 ml filter-sterilized standard vitamin solution (see below).The final pH of this medium should be 6.8 without adjustment. | |||||
| 2 For chemolithotrophic growth incubate the culture under an atmosphere of 2% (v/v) O2, 10% CO2, 60% H2 and 28% N2. For heterotrophic growth supplement the mineral medium with an appropriate carbon source (0.2% carbohydrate or 0.1% organic acid). For growth on nitrogen-free medium, omit NH4Cl and incubate the culture under an atmosphere of 2% (v/v) O2, 10% CO2, 10% H2 and 78% N2 or heterotrophically under 2% (v/v) O2 and 98% N2. For more details see Ref. 1515 and Ref. 3363. | |||||
Solution B #
925
| Compound | Amount | Unit | |
|---|---|---|---|
|
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NH4Cl | 1.00 | g |
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MgSO4 x 7 H2O | 0.50 | g |
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CaCl2 x 2 H2O | 0.01 | g |
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MnCl2 x 4 H2O | 5.00 | mg |
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NaVO3 x H2O | 5.00 | mg |
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Trace element solution SL-6 | 5.00 | ml |
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Distilled water | 920.00 | ml |
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Agar , if required | 20.00 | g |
Standard vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
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Riboflavin | 10.0 | mg |
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Thiamine-HCl x 2 H2O | 50.0 | mg |
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Nicotinic acid | 50.0 | mg |
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Pyridoxine hydrochloride | 50.0 | mg |
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Calcium pantothenate | 50.0 | mg |
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Biotin | 0.1 | mg |
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Folic acid | 0.2 | mg |
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Vitamin B12 | 1.0 | mg |
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Distilled water | 100.0 | ml |
Trace element solution SL-6 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnSO4 x 7 H2O | 0.10 | g |
|
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MnCl2 x 4 H2O | 0.03 | g |
|
|
H3BO3 | 0.30 | g |
|
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CoCl2 x 6 H2O | 0.20 | g |
|
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CuCl2 x 2 H2O | 0.01 | g |
|
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NiCl2 x 6 H2O | 0.02 | g |
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Na2MoO4 x 2 H2O | 0.03 | g |
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Distilled water | 1000.00 | ml |