培養基配方
Final pH: 6.5 - 7.0
Final volume: 1005 ml
NH4Cl 0.50 g
MgSO4 x 7 H2O 0.50 g
CaCl2 x 2 H2O 0.25 g
NaCl 2.25 g
FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4) 2.00 ml
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Yeast extract 2.00 g
Casitone 2.00 g
Sodium resazurin (0.1% w/v) 0.50 ml
K2HPO4 0.35 g
KH2PO4 0.23 g
Na2CO3 1.00 g
D-Fructose 5.00 g
Wolin's vitamin solution (10x) 1.00 ml
L-Cysteine HCl x H2O 0.30 g
Na2S x 9 H2O 0.30 g
Distilled water 1000.00 ml
Dissolve ingredients (except phosphates, carbonate, fructose, vitamins, cysteine and sulfide) and 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. After autoclaving add phosphates, fructose, vitamins, cysteine and sulfide to the medium 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 fructose and vitamins should be sterilized by filtration. Adjust pH of complete medium to pH 6.5 - 7.0, if necessary.
For DSM 4440: Replace fructose with 1.35 g/l betaine as substrate; cysteine and sulfide must be replaced by 0.15 g/l DL-dithiothreitol (DTT) added from an anoxic stock solution sterilized by filtration.
For DSM 6539, DSM 6540, DSM 17108, DSM 116159, DSM 116160: Replace fructose with
2.00 g/l D-glucose added to the autoclaved medium from a filter-sterilized anoxic stock
solution. Cysteine and sulfide must be replaced by 0.15 g/l DL-dithiothreitol (DTT) added
from an anoxic stock solution sterilized by filtration.
For DSM 11501: Supplement medium with 0.30 g DL-Dithiothreitol and 20 ml/l methanol
(50% v/v). Omit D-fructose, Na2S x 9 H2O, and L-Cysteine HCl x H2O. After inoculation, add
1 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.
For DSM 14980: Replace fructose with 8.00 g/l D-glucose and 2.00 g/l Na-pyruvate as
substrates added to the autoclaved medium from filter-sterilized anoxic stock solutions.
For DSM 16652: Replace fructose with 1.11 g/l N-acetyl-D-glucosamine as substrate added
to the autoclaved medium from a filter-sterilized anoxic stock solution.
For DSM 17189, DSM 17285: Replace fructose with 3.60 g/l D-mannitol as substrate added
to the autoclaved medium from a filter-sterilized anoxic stock solution. Cysteine and
sulfide must be replaced by 0.30 g/l DL-dithiothreitol (DTT) added from an anoxic stock
solution sterilized by filtration.
For DSM 26537: Replace fructose with 5.00 g/l D-glucose as substrate added to the autoclaved medium from a filter-sterilized anoxic stock solution. Adjust pH of completed medium to 7.8 using a sterile anoxic stock solution of 5% (w/v) Na2CO3.
For DSM 26827: Replace fructose with 2.50 g/l Na-(DL)-lactate added to the autoclaved medium from a separately sterilized anoxic stock solution. Cysteine and sulfide must be replaced by 0.15 g/l DL-dithiothreitol (DTT) added from an anoxic stock solution sterilized by filtration.
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.
Selenite-tungstate solution (from medium 385)
NaOH 0.50 g
Na2SeO3 x 5 H2O 3.00 mg
Na2WO4 x 2 H2O 4.00 mg
Distilled water 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
