KCTC Media No. 334 : Acetobacterium Medium

조성	용량	단위
NH4Cl	1.0	g
KH2PO4	0.33	g
MgSO4·7H2O	0.10	g
K2HPO4	0.45	g
Trace element solution (see below)	20.0	ml
조성 용량 단위 Nitrilotriacetic acid 1.50 g MgSO4 . 7H2O 3.0 g MnSO4 . H2O 0.5 g NaCl 1.0 g FeSO4 . 7H2O 0.1 g CoSO4 . 7H2O 0.18 g ZnSO4 . 7H2O 0.18 g CaCl2 . 2H2O 0.1 g CuSO4 . 5H2O 0.01 g KAl(SO4)2 . 12H2O 0.02 g H3BO3 0.01 g Na2MoO4 . 2H2O 0.01 g NiCl2 . 6H2O 0.03 g Na2SeO3 . 5H2O 0.3 mg Distilled water 1.0 L First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final pH 7.0 (with KOH).
Vitamin solution (see below)	10.0	ml
조성 용량 단위 Biotin 2.0 mg Folic acid 2.0 mg Pyridoxine-HCl 10.0 mg Thiamine-HCl . 2H2O 5.00 mg Riboflavin 5.0 mg Nicotinic acid 5.0 mg D-Ca-pantothenate 5.0 mg Vitamin B12 0.10 mg p-Aminobenzoic acid 5.0 mg Lipoic acid 5.0 mg Distilled water 1.0 L
Yeast extract	2.0	g
Fructose	10.0	g
Resazurin solution (0.1%w/v)	0.5	ml
NaHCO3	10.0	g
Cysteine-HCl·H2O	0.5	g
Na2S·9H2O	0.5	g
Distilled water	1.0	L

** ph : -

** memo : Trace element solution (KCTC Media no.918)
Vitamin solution (KCTC Media no.1335)

** 추가조치 사항 : Dissolve ingredients except NaHCO_3_, fructose, cysteine and sodium sulfide, bring to the
boil for few minutes and cool to room temperature under N_2_ + CO_2_ (80 + 20) gas
mixture. Add NaHCO_3_ (solid) and equilibrate the medium with the gas until a pH of c. 7.4
is reached. Then distribute and autoclave under the same gas. Before use adjust the pH
to 8.2 by adding sterile anaerobic Na_2_CO_3_ solution (c. 0.25 ml of 5% Na_2_CO_3_ per 10 ml
medium) and add fructose, cysteine and sodium sulfide from anaerobic sterile stock
solutions.
Note: For autotrophic growth fructose is omitted and a gas atmosphere of 80% H_2_ + 20% CO_2_ is used.

For KCTC 5016 use only 1.0 g/l of NaHCO_3_ and omit Na_2_CO_3_ to reach a final pH of 6.5.
In addition, add per liter 1 ml from following solution: Na_2_SeO_3_ . 5H_2_O, 3.0 mg; Na_2_WO_4_ . 2H_2_O, 4 mg; NaOH, 0.5 g; 1 l distilled water.
Mix all of the ingredients with exception of fructose. Equilibrate medium with 80 % N_2_ and 20 % CO_2_ gas mixture until a pH of 7.4 is reached. Tube and autoclave under the same environment. Before inoculation adjust the pH to 8.2 by adding sterile anaerobic Na_2_CO_3_(0.25 ml of 5 % Na_2_CO_3_ per 10 ml medium)
and add sterile solution of fructose to a final concentration of 1 %. For autotrophic growth fructose is omitted and gas atmosphere of 80 % H_2 _and 20 % CO_2_ is used.