培養基配方
NH4Cl 0.33 g
KH2PO4 0.33 g
KCl 0.33 g
MgCl2 x 6 H2O 0.33 g
CaCl2 x 2 H2O 0.33 g
Trace element solution SL-10 1.00 ml
Yeast extract 0.50 g
Sodium resazurin (0.1% w/v) 0.50 ml
Na2CO3 1.50 g
Cellobiose 5.00 g
Wolin's vitamin solution (10x) 1.00 ml
Na2S x 9 H2O 0.50 g
Distilled water 1000.00 ml
Dissolve ingredients (except vitamins, carbonate, cellobiose and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add vitamins, cellobiose and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of vitamins and cellobiose should be sterilized by filtration. The pH of the complete medium should be at 7.1 - 7.3.
For DSM 6724: Replace cellobiose with 5.00 g/l soluble starch as substrate added to the autoclaved medium from a sterile anoxic stock solution.
For DSM 9003: Supplement medium with 1.00 g/l Trypticase peptone and increase amount of yeast extract to 2.00 g/l.
Trace element solution SL-10 (from medium 320)
HCl (25%) 10.00 ml
FeCl2 x 4 H2O 1.50 g
ZnCl2 70.00 mg
MnCl2 x 4 H2O 100.00 mg
H3BO3 6.00 mg
CoCl2 x 6 H2O 190.00 mg
CuCl2 x 2 H2O 2.00 mg
NiCl2 x 6 H2O 24.00 mg
Na2MoO4 x 2 H2O 36.00 mg
Distilled water 990.00 ml
First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally
make up to 1000.00 ml.
Wolin's vitamin solution (10x) (from medium 120)
Biotin 20.00 mg
Folic acid 20.00 mg
Pyridoxine hydrochloride 100.00 mg
Thiamine HCl 50.00 mg
Riboflavin 50.00 mg
Nicotinic acid 50.00 mg
Calcium D-(+)-pantothenate 50.00 mg
Vitamin B12 1.00 mg
p-Aminobenzoic acid 50.00 mg
(DL)-alpha-Lipoic acid 50.00 mg
Distilled water 1000.00 ml
