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843. TREPONEMA AZOTONUTRICIUM MEDIUM

Culture medium recipe

Vol.
×
  • For replace sugar mix with 1.00 g/l xylan as substrate.
Equipment needed: Autoclave Filter Hungate tubes
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.00 g 0.996 17.043
2 0.50 g 0.498 6.68
3 0.40 g 0.398 1.96
4 0.10 g 0.1 0.677
5 0.30 g 0.299 5.586
6 0.20 g 0.199 1.464
7 0.15 g 0.149 1.052
8 Trace element solution SL-10 1.00 ml - -
9 Selenite-tungstate solution 1.00 ml - -
10
(BD BBL)
1.00 g 0.996 -
11
(0.1% w/v)
0.50 ml 5.0e-4 0.002
12 1.50 g 1.494 14.096
13 Sugar mix 30.00 ml - -
14 Yeast autolysate 50.00 ml - -
15 Seven vitamins solution 1.00 ml - -
16 Co-factors solution 20.00 ml - -
17 0.40 g 0.398 2.583
18 900.00 ml 894.807 49669.3
19 , might need strain adjustment 1.00 g/l - -
Instructions:
  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.
  2. Check pH of complete medium and adjust to 7.2 - 7.4, if necessary.
  3. Note: Basal medium and yeast autolysate have to be prepared freshly and cannot be stored for longer periods.
Solution occurs in 1 other media (first defined in medium 843 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.70 g 56.667 165.549
2 1.70 g 56.667 165.548
3 1.80 g 60 166.523
4 1.90 g 63.333 185.025
5 0.75 g 25 166.522
6 30.00 ml 998.207 55408.8
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1
(SIGMA YSC-1)
14 g 280 -
2 50 ml 998.207 55408.8
Instructions:
  1. Suspend 14.0 g of dry baker's yeast (SIGMA YSC-1) in 50 ml 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.
Equipment needed: Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 2.50 g 2.5 5.426
2 0.25 g 0.25 1.228
3 0.25 g 0.25 1.012
4 0.05 g 0.05 0.098
5 0.05 g 0.05 0.075
6 0.05 g 0.05 0.065
7 0.05 g 0.05 0.064
8 65.00 mg 0.065 -
9 25.00 mg 0.025 0.205
10 2.50 mg 0.003 0.006
11 0.50 mg 5.0e-4 0.001
12 1000.00 ml 998.207 55408.8
Instructions:
  1. Adjust pH to 7.0 with 1 N NaOH against indicator paper, filter sterilize and store under 100% N2 gas atmosphere.
Solution occurs in 161 other media (first defined in medium 385 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.5 g 0.5 12.501
2 3.0 mg 0.003 0.011
3 4.0 mg 0.004 0.012
4 1000.0 ml 998.207 55408.8
Solution occurs in 86 other media (first defined in medium 503 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 100 mg 0.1 0.063
2 80 mg 0.08 0.583
3 20 mg 0.02 0.082
4 200 mg 0.2 1.625
5 100 mg 0.1 0.21
6 300 mg 0.3 1.459
7 200 mg 0.2 0.536
8 1000 ml 998.207 55408.8
Solution occurs in 239 other media (first defined in medium 320 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2
(25%; 7.7 M)
10.0 ml 3.17 86.943
3 1.5 g 1.5 7.545
4 70.0 mg 0.07 0.514
5 100.0 mg 0.1 0.505
6 6.0 mg 0.006 0.097
7 190.0 mg 0.19 0.799
8 2.0 mg 0.002 0.012
9 24.0 mg 0.024 0.101
10 36.0 mg 0.036 0.149
11 990.0 ml 988.225 54854.8
Instructions:
  1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 ml.

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.3
Equipment needed: Autoclave, Filter, Hungate tubes

Associated strains

All strains for this medium: 2
Strains with modifications: 2

Gas composition

  • 100 % N2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 997.203 55353.1
Baker's yeast 13.944 -
Trehalose 1.892 5.529
Maltose x H2O 1.793 4.976
Cellobiose 1.693 4.947
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Cultivation metadata from 2 strains BacDive

pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain

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