培養基配方
Final pH: 6.7 - 7.0
Final volume: 1011 ml
Ferric citrate (Aldrich F6129) 13.70 g
NH4Cl 1.50 g
NaH2PO4 0.60 g
KCl 0.10 g
Na-acetate 2.50 g
Modified Wolin's mineral solution 10.00 ml
NaHCO3 2.50 g
Wolin's vitamin solution (10x) 1.00 ml
Distilled water 1000.00 ml
1. First dissolve ferric citrate in water by heating and adjust to pH 6.0, then add other medium ingredients except bicarbonate and vitamins. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add and dissolve bicarbonate and adjust pH to 6.8. Thereafter, dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins to the sterile medium from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 6.7 - 7.0, if necessary.
2. Note: Use 5 - 10% (v/v) inoculum.
Modified Wolin's mineral solution (from medium 141)
Nitrilotriacetic acid 1.50 g
MgSO4 x 7 H2O 3.00 g
MnSO4 x H2O 0.50 g
NaCl 1.00 g
FeSO4 x 7 H2O 0.10 g
CoSO4 x 7 H2O 0.18 g
CaCl2 x 2 H2O 0.10 g
ZnSO4 x 7 H2O 0.18 g
CuSO4 x 5 H2O 0.01 g
AlK(SO4)2 x 12 H2O 0.02 g
H3BO3 0.01 g
Na2MoO4 x 2 H2O 0.01 g
NiCl2 x 6 H2O 0.03 g
Na2SeO3 x 5 H2O 0.30 mg
Na2WO4 x 2 H2O 0.40 mg
Distilled water 1000.00 ml
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.
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
