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조성 | 용량 | 단위 |
---|---|---|
KH2PO4 | 0.2 | g |
** ph : -
** memo : KH2PO4 0.2 g
NH4Cl 0.25 g
NaCl 20 g
MgCl2·6H2O 3 g
KCl 0.5 g
CaCl2·2H2O 0.15 g
Trace element mixture* 1 ml
Selenite-tungstate solution** 1 ml
Vitamin mixture*** 1 ml
Vitamin B12 solution**** 1 ml
Bicarbonate solution (1.0 M)***** 30 ml
Dithionite (ca.) 20 mg
Distilled water 1 L
Mix ingredients except vitamin mixture, vitamin B12 solution, bicarbonate solution and dithionite, and adjust pH to about 7.5. Dispense the medium into suitable culture vessels under a stream of nitrogen gas, seal with butyl rubber stoppers, and autoclave at 121°C for 15 min. Aseptically and anaerobically add vitamin mixture, vitamin B12 solution, bicarbonate solution and dithionite solution using sterile syringes, and the N2/CO2 (80/20) mixed gas is enclosed with the vessels. Adjust pH to 7.2- 7.8 using compact pH meter. An aqueous solution of sodium glutamate (final concentration, 10 mM) and sodium thiosulfate (final concentration, 20 mM) are added before inoculation. Addition of yeast extract (0.5 - 1 g/L) greatly improves the growth of the strain.
*Trace element mixture
HCl (25%) 12.5 ml
FeSO4·7H2O 2100 mg
H3BO3 30 mg
MnCl2·4H2O 100 mg
CoCl2·6H2O 190 mg
NiCl2·6H2O 24 mg
CuCl2·2H2O 2 mg
ZnSO4·7H2O 144 mg
Na2MoO4·2H2O 36 mg
Distilled water 987 ml
Sterilized by autoclaving.
**Selenite-tungstate solution
NaOH 0.4 g
Na2SeO·5H2O 6 mg
Na2WO4·2H2O 8 mg
Distilled water 1 L
Sterilized by autoclaving.
***Vitamin mixture
4-Aminobenzoic acid 4 mg
D(+)-Biotin 1 mg
Nicotinic acid 10 mg
Calcium D(+)-panthothenate 5 mg
Pyridoxine dihydrochloride 15 mg
Thiamine-HCl 10 mg
Distilled water 100 ml
Sterilized by filtration.
****Vitamin B12 solution
Cyanocobalamin 5 mg
Distilled water 100 ml
Sterilized by filtration.
*****Bicarbonate solution (1.0 M)
NaHCO3 84 g
Distilled water 1 L
Sterilized by filtration.
** 추가조치 사항 : -