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JCM 411 . Fe(III)-REDUCER MARINE ENRICHMENT MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 19.000 g 950 16256
MgCl2 x 6 H2O 9.000 g 450 2213.45
MgSO4 x 7 H2O 0.150 g 7.5 30.429
CaCl2 x 2 H2O 0.300 g 15 102.03
KCl 0.500 g 25 335.345
KH2PO4 0.420 g 21 154.314
(NH4)2SO4 0.100 g 5 37.839
NaBr 0.050 g 2.5 24.298
SrCl2 x 6 H2O 0.020 g 1 3.751
Yeast extract
(BD-Difco)
0.100 g 5 -
NaHCO3 2.500 g 125 1487.97
Na2SeO4 0.020 g 1 5.292
Poorly crystalline iron(III) oxide 100.000 mM - -
Trace vitamins 10.000 ml - -
Trace mineral solution 10.000 ml - -
L-Cysteine HCl x H2O 0.044 g 2.2 -
FeCl2 x 4 H2O 0.260 g 13 65.388
1 Add components except L-cysteine x HCl x H2O and FeCl2 x 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 x HCl x H2O and FeCl2 x 4H2O solutions under a N2 gas atmosphere. Prior to inoculation, add the sterile L-cysteine x HCl x H2O and FeCl2 x 4H2O solutions. Pressurize the inoculated vessels to 100 kPa H2-CO2 (4:1, v/v) and cultivate without shaking.
Solution occurs in 3 other media (first defined in Medium J411).
Equipment needed: Centrifuge
1 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).
Solution occurs in 4 other media (first defined in Medium J411).
1 Use Trace minerals of Medium No. 151 supplemented with 0.025 g (final) Na2MoO4 x 2H2O, 0.024 g NiCl2 x 6H2O and 0.025 g Na2WO4 x 2H2O (per liter).
2 Comment: For strain JCM 12378, inoculate with a 10% inoculum.
Solution occurs in 220 other media (first defined in Medium J197).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 -
Thiamine HCl 5.0 mg 0.005 -
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium pantothenate 5.0 mg 0.005 0.01
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Media status: Inofficial medium
Source: JCM
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Centrifuge

Associated strains

All strains for this medium: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 950 16256
MgCl2 x 6 H2O 450 2213.45
NaHCO3 125 1487.97
KCl 25 335.345
KH2PO4 21 154.314
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