Medium data
Search for medium no. = [411].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
411 Fe(III)-REDUCER MARINE ENRICHMENT MEDIUM
| NaCl | 19.0 | g |
| MgCl2·6H2O | 9.0 | g |
| MgSO4·7H2O | 0.15 | g |
| CaCl2·2H2O | 0.3 | g |
| KCl | 0.5 | g |
| KH2PO4 | 0.42 | g |
| (NH4)2SO4 | 0.1 | g |
| NaBr | 0.05 | g |
| SrCl2·6H2O | 0.02 | g |
| Yeast extract (BD-Difco) | 0.1 | g |
| NaHCO3 | 2.5 | g |
| Na2SeO4 | 0.02 | g |
| Poorly crystalline iron(III) oxide (see below) | 100.0 | mM |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
| Trace mineral solution (see below) | 10.0 | ml |
| L-Cysteine·HCl·H2O | 0.044 | g |
| FeCl2·4H2O | 0.26 | g |
Add components except L-cysteine·HCl·H2O and FeCl2·4H2O to distilled water, bring volume to 1.0 L and adjust pH to 7.0. Distribute the medium into appropriate culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a H2-CO2 (4:1, v/v) gas mixture, sparge with the same gas mixture for 10 - 15 min, seal with butyl rubber stoppers and autoclave. Separately autoclave concentrated (100- or 200-fold) L-cysteine·HCl·H2O and FeCl2·4H2O solutions under a N2 gas atmosphere. Prior to inoculation, add the sterile L-cysteine·HCl·H2O and FeCl2·4H2O solutions. Pressurize the inoculated vessels to 100 kPa H2-CO2 (4:1, v/v) and cultivate without shaking.
Poorly crystalline iron(III) oxide:
Dissolve 108 g FeCl3 in 1.0 L distilled water. Adjust pH to 7.0 with 10 N NaOH. Allow the suspension to sit for 30 min, then readjust pH to 7.0. Centrifuge the suspension at 5,000 x g for 20 min and pour off supernatant. Wash the precipitate with distilled water by resuspending in distilled water followed by the centrifugation. Repeat the washing at least 5 times, then, resuspend in 1.0 L distilled water. Use the Fe(III) suspension as 0.67 M solution. More precisely, check the concentration of Cl- to be less than 1 mM and determine the concentration of Fe(III) by using hydroxylamine as described by Lovley and Phillips (Appl. Environ. Microbiol. 54, 751-757, 1986).
Trace mineral solution:
Use Trace minerals of Medium No. 151 supplemented with 0.025 g (final) Na2MoO4·2H2O, 0.024 g NiCl2·6H2O and 0.025 g Na2WO4·2H2O (per liter).
Comment: For strain JCM 12378, inoculate with a 10% inoculum.
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