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DSMZ

DSMZ 843 . HELMUTKOENIGIA MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 843 #

ml
Equipment needed: Hungate tubes Filter Centrifuge Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 0.997 17.06
KCl 0.50 g 0.499 6.687
MgCl2 x 6 H2O 0.40 g 0.399 1.962
CaCl2 x 2 H2O 0.10 g 0.1 0.678
NH4Cl 0.30 g 0.299 5.592
KH2PO4 0.20 g 0.199 1.465
Na2SO4 0.15 g 0.15 1.053
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Trypticase peptone
(BD BBL) 		1.00 g 0.997 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.496 14.11
Sugar mix 30.00 ml - -
Yeast autolysate 50.00 ml - -
Seven vitamins solution 1.00 ml - -
Co-factors solution 20.00 ml - -
DL-Dithiothreitol
(DTT) 		0.40 g 0.399 2.585
Distilled water 900.00 ml - -
1 Dissolve ingredients (except carbonate, sugar mix, yeast autolysate, vitamins, co-factors, and DTT), sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume (e.g., 4.5 ml per Hungate-type tube) and autoclave. After sterilization add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere and sugar mix, yeast autolysate, vitamins, co-factors and DTT from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and sterilized by filtration. Check pH of complete medium and adjust to 7.2 - 7.4, if necessary.
2 Note: Basal medium and yeast autolysate have to be prepared freshly and cannot be stored for longer periods.

Sugar mix #

1000
Solution occurs in 1 other media (first defined in Medium 843).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Cellobiose 56.7 g 56.7 165.644
Sucrose 56.7 g 56.7 165.644
Maltose x H2O 60.0 g 60 175.287
Trehalose 63.3 g 63.3 184.926
D-Xylose 25.0 g 25 166.522
Distilled water 1000.0 ml - -

Yeast autolysate #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract
(dry baker's yeast, SIGMA YSC-1) 		280 g 280 -
Distilled water 1000 ml - -
1 Suspend dry baker's yeast (SIGMA YSC-1) in distilled water and allow cells to autolyse for 48 h at 55°C. Thereafter, cell debris are removed by centrifugation (3.700 x g for 30 min) and the amber-colored supernatant is stored in aliquots at -20°C.

Co-factors solution #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
Thiamine pyrophosphate 2.50 g 2.5 5.426
Pyridoxal hydrochloride 0.25 g 0.25 1.228
Pyridoxal phosphate 0.25 g 0.25 1.012
Ca-folinate 0.05 g 0.05 0.098
ß-NAD 0.05 g 0.05 -
Coenzyme A 0.05 g 0.05 0.065
FAD 0.05 g 0.05 0.064
Haemin
(dissolved separately in 10 mM NaOH) 		65.00 mg 0.065 0.1
Nicotinamide 25.00 mg 0.025 0.205
Folic acid 2.50 mg 0.003 0.006
Riboflavin 0.50 mg 5.0e-4 0.001
Distilled water 1000.00 ml - -
1 Adjust pH to 7.0 with 1 N NaOH against indicator paper, filter sterilize and store under 100% N2 gas atmosphere.

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.

Selenite-tungstate solution #

1000
Solution occurs in 179 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -

Seven vitamins solution #

1000
Solution occurs in 78 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 25.10.22
Source: DSMZ
Synonyms: TREPONEMA AZOTONUTRICIUM MEDIUM
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.2 - 7.4
Equipment needed: Autoclave, Centrifuge, 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: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Yeast extract 13.958 -
Trehalose 1.893 5.531
Maltose x H2O 1.795 5.243
Sucrose 1.696 4.954
Cellobiose 1.696 4.954
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Cultivation metadata from 2 strains BacDive

pH 6 - 8 1 strain
phyla 2 2 strains
oxygen anaerobe 1 strain
temperature mesophilic 1 strain