# J741: ACETOBACTERIUM MEDIUM


## Main sol. J741

1	g	NH4Cl	
0.33	g	KH2PO4	
0.45	g	K2HPO4	
0.1	g	MgSO4 x 7 H2O	
20	ml	**Trace element solution**	
20	ml	**Trace vitamins**	
2	g	Yeast extract	
10	g	Fructose	
1	mg	Resazurin	
10	g	NaHCO3	
0.5	g	L-Cysteine HCl x H2O	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except NaHCO3, fructose, cysteine and sodium sulfide, bring to a boil for a few minutes and cool to room temperature under a N2-CO2 (80:20, v/v) gas mixture. Add NaHCO3 (solid) and equilibrate the medium with the gas until pH reaches ca. 7.4. Then distribute and autoclave under the same gas atmosphere. Before use, adjust pH to 8.2 by adding sterile anaerobic Na2CO3 solution (ca. 0.25 ml of 5% Na2CO3 per 10 ml medium) and add fructose, cysteine and sodium sulfide from anaerobic sterile stock solutions.


## Trace element solution

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x n H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
10	mg	CuSO4 x 5 H2O	
20	mg	AlK(SO4)2 x 12 H2O	
10	mg	H3BO3	
10	mg	Na2MoO4 x 2 H2O	
25	mg	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid at first and adjust pH to 6.5 with KOH, then add the remaining minerals.
2. Adjust final pH of the medium to 7.0 with KOH.


## Trace vitamins

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water