培養(yǎng)基配方
NH4Cl 1.00 g
Na2SO4 2.00 g
Na2S2O3 x 5 H2O 1.00 g
MgSO4 x 7 H2O 1.00 g
CaCl2 x 2 H2O 0.10 g
KH2PO4 0.50 g
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Yeast extract 1.00 g
Sodium resazurin (0.1% w/v) 0.50 ml
Na2CO3 1.00 g
Na-DL-lactate 2.50 g
Wolin's vitamin solution (10x) 1.00 ml
Na2S x 9 H2O 0.10 g
Distilled water 1000.00 ml
1. Dissolve ingredients (except carbonate, vitamins, lactate and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving complete the medium by adding vitamins (sterilized by filtration), lactate 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 atmosphere. Adjust pH of the complete medium to 7.0 - 7.2, if necessary.
2. Note: Prior to inoculation 10-20 mg/l sodium dithionite (added from a 5% w/v solution freshly prepared under N2 and filter-sterilized) can be added to the medium to stimulate growth at the beginning.
For DSM 7073, DSM 101608, DSM 101609: Replace lactate with 0.9 g/l glycerol and adjust pH of complete medium to 7.5.
For DSM 10017: Replace lactate with 2.5 g/l Na-pyruvate added to the autoclaved medium from an anoxic stock solution sterilized by filtration and replace sodium sulfide with 0.15g/l DL-dithiothreitol (DTT) added from an anoxic stock solution sterilized by filtration.
For DSM 10349: Replace lactate with 2.5 g/l Na-pyruvate added to the autoclaved medium from an anoxic stock solution sterilized by filtration and supplement medium with an additional amount of 1.0 g/l Na2S2O3 x 5 H2O added from an anoxic stock solution sterilized by filtration.
For DSM 13257, DSM 16036, DSM 16058, DSM 24088, DSM 24093, DSM 24523, DSM 26885: Replace lactate with 2.5 g/l Na-pyruvate added to the autoclaved medium from an anoxic stock solution sterilized by filtration.
For DSM 18151: Replace lactate with 0.2 g/l Na-acetate. After inoculation pressurize culture vessels to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
For DSM 21394: Replace lactate with 5.0 ml/l methanol added to the autoclaved medium from an anoxic stock solution prepared under 100% N2
gas.
For DSM 24089, DSM 102940: Replace lactate with 3.5 g/l D-fructose added to the autoclaved medium from an anoxic stock solution sterilized by filtration.
For DSM 25471: Replace lactate with 2.0 g/l D-glucose added to the autoclaved medium from a sterile anoxic stock solution.
For DSM 28127: Replace lactate with 0.4 g/l Na-benzoate added to the autoclaved medium from a sterile anoxic stock solution.
For DSM 111569: Supplement medium with 2.20 g/l pyruvate, 1.20 g/l taurine and 0.20 mg/l 1,4-naphthochinone added to the autoclaved medium from anoxic stock solutions 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
