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DSMZ

DSMZ 520 . RUMINOCLOSTRIDIUM CELLULOLYTICUM (CM3) MEDIUM

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
×

Main sol. 520 #

ml
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.296 9.809
KH2PO4 1.50 g 1.496 10.99
K2HPO4 x 3 H2O 2.90 g 2.891 16.6
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		1.25 ml 0.001 0.004
Trace element solution SL-10 1.00 ml - -
Yeast extract 2.00 g 1.994 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
MgCl2 x 6 H2O 0.20 g 0.199 0.981
CaCl2 x 2 H2O 75.00 mg 0.075 0.509
Cellobiose 6.00 g 5.982 17.476
Cellulose , MN 301
optional 		10.00 g 9.97 -
Na2CO3 1.50 g 1.496 14.11
L-Cysteine HCl x H2O 0.50 g 0.499 2.838
Distilled water 1000.00 ml - -
1 Dissolve ingredients except magnesium chloride, calcium chloride, cellobiose, cysteine and carbonate, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add magnesium chloride, calcium chloride and cellobiose from anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2. Cellobiose has to be sterilized by filtration. Prior to inoculation add cysteine from a sterile anoxic stock solution prepared under 100% N2 gas and adjust pH to 7.2.
2 Note: Some strains can be adapted to cellulose as substrate using 10.00 g/l cellulose powder MN 301 (MACHEREY-NAGEL).

Trace element solution SL-10 #

1000
Solution occurs in 266 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Metadata

Last modified: 04.04.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.2
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % N2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 15
Strains with modifications: 4

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Cellulose
optional 		9.97 -
Cellobiose 5.982 17.476
K2HPO4 x 3 H2O 2.891 16.6
Yeast extract 1.994 -
Na2CO3 1.496 14.11
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Cultivation metadata from 15 strains BacDive

pH 6 - 8 2 strains
pH 8 - 10 1 strain
pH 4 - 6 1 strain
phyla 1 13 strains
nutrition chemoorganotroph 1 strain
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