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DSMZ

DSMZ 1681 . Mineral salt medium with colloidal chitin

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

Culture medium recipe

Vol.
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Medium for Paludihabitans psychrotolerans DSM 104738

This strain uses the standard medium without modifications.

Growth conditions: 28 °C

Main sol. 1681 #

ml
Equipment needed: Filter Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 50.00 ml - -
Solution B 50.00 ml - -
Modified Wolin's mineral solution 10.00 ml - -
Wolin's vitamin solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		1.00 ml 1.0e-3 -
Yeast extract 1.00 g 1 -
Polypeptone 10.00 g 10 -
NaHCO3 3.52 g 3.52 41.901
Colloidal chitin 5.00 g 5 -
Distilled water 890.00 ml - -
1 Reducing agents: Flush each solution under N2 for 30 minutes, stopper and autoclave. L-Cysteine-HCL x H2O solution (10%): add 12.5 µl per 10 ml medium Na2S x 9 H2O solution (3%): add 40 µl per 10 ml medium.
2 Colloidal chitin preparation: Cut 5 gram commercial crab (shrimp) shell flakes into small pieces, and dissolve in 100 ml of 12M HCL while stirring in a fume hood at room temperature. Add chitin-HCL mixture slowly into pre-cooled water to obtain the colloidal chitin, centrifuge and wash in pre-cooled water till the colloidal chitin reaches a pH of 6 to 6.5. Mix 0.5% with MS medium as the sole carbon source. As an alternative commercial available chitin powder can be used.
3 Prepare MS medium with chitin: Dissolve the ingredients of MS medium (except NaHCO3 and vitamins), boil medium under CO2 until the resazurin indicator changes from pink to colourless. Cool down under CO2, add NaHCO3 and adjust pH to 7.0. Switch to N2 to prevent further lowering of the pH. Fill up into Hungate tubes under N2 and autoclave. Before use add vitamins (sterilized by filtration) and reducing agents.

Solution A #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 8 g 8 58.787
NH4Cl 5 g 5 93.475
MgCl2 x 6 H2O 4 g 4 19.675
CaCl2 x 2 H2O 1 g 1 6.802
Distilled water 1000 ml - -

Solution B #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 8 g 8 45.931
Distilled water 1000 ml - -

Wolin's vitamin solution #

1000
Solution occurs in 53 other media (first defined in Medium 141).
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 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 0.021
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Modified Wolin's mineral solution #

1000
Solution occurs in 166 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x H2O 0.50 g 0.5 2.958
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.18 g 0.18 1.161
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.18 g 0.18 0.626
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 x 12 H2O 0.02 g 0.02 0.042
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2SeO3 x 5 H2O 0.30 mg 3.0e-4 0.001
Na2WO4 x 2 H2O 0.40 mg 4.0e-4 0.001
Distilled water 1000.00 ml - -
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.

Metadata

Last modified: 27.04.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Associated strains

All strains for this medium: 1
Paludihabitans psychrotolerans DSM 104738

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Polypeptone 10 -
Colloidal chitin 5 -
NaHCO3 3.52 41.901
Yeast extract 1 -
KH2PO4 0.4 2.939
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Cultivation metadata from 1 strains BacDive

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