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DSMZ 320 . CLOSTRIDIUM CELLULOVORANS MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Filter Gassing station Hungate tubes Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 x 3 H2O 1.00 g 1 5.741
NH4Cl 1.00 g 1 18.695
KCl 0.50 g 0.5 6.707
MgSO4 x 7 H2O 0.50 g 0.5 2.029
Trypticase peptone
(BD BBL)
0.50 g 0.5 -
Yeast extract 0.50 g 0.5 -
Clarified rumen fluid 20.00 ml - -
Sludge fluid , alternative 20.00 ml - -
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
L-Cysteine HCl x H2O 0.15 g 0.15 0.854
Na2CO3 1.00 g 1 9.435
Cellobiose 5.00 g 5 14.607
Na2S x 9 H2O 0.15 g 0.15 0.625
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except cysteine, carbonate, cellobiose and sulfide), bring medium to the boil, then cool to room temperature under 80% N2 and 20% CO2 gas mixture and add cysteine. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Sterilize cellobiose separately by filtration under 100% N2 gas. Adjust pH of the complete medium to 7.0, if necessary.
2 Note: Some strains can be adapted to cellulose as substrate using 10.0 g/l cellulose powder MN 301 (MACHEREY-NAGEL).
Solution occurs in 262 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.
Solution occurs in 8 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 0.4 g 4 -
Sludge 100.0 ml - -
1 Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at room temperature in the dark.
Solution occurs in 27 other media (first defined in Medium 1310).
1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 minand then centrifuged at 27,000 g for 20 min The supernatant is made anoxic by sparging with 100% N2 gas for 15 min dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes

Gas composition

  • 20 % CO2
  • 80 % N2

Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Cellobiose 5 14.607
NH4Cl 1 18.695
Na2CO3 1 9.435
K2HPO4 x 3 H2O 1 5.741
KCl 0.5 6.707
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Cultivation metadata from 1 strains BacDive

pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
oxygen obligate anaerobe 1 strain
temperature mesophilic 1 strain

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