# J187: DESULFUROCOCCUS AMYLOLYTICUS MEDIUM


## Main sol. J187

0.33	g	NH4Cl	
0.33	g	KH2PO4	
0.33	g	KCl	
0.44	g	CaCl2 x 2 H2O	
0.7	g	MgCl2 x 6 H2O	
0.5	g	NaCl	
0.8	g	NaHCO3	
1	ml	**FeCl2 solution**	
1	ml	**Trace element solution**	
10	ml	**Trace vitamins**	
0.2	g	Yeast extract (BD-Difco)	
5	g	Bacto peptone (BD-Difco)	
10	g	Sulfur (powder)	
0.5	g	Na2S x 9 H2O	
1	mg	Resazurin	
990	ml	Distilled water	

1. Mix ingredients, except yeast extract, peptone, vitamins, sulfur and Na2S x 9H2O, and autoclave under a N2-CO2 (80:20) atmosphere. Separately autoclave yeast extract and peptone as 10% solutions and Na2S x 9H2O as a 5% solution under a N2 atmosphere. Steam sulfur for 3 hr on each of 3 successive days. Filter-sterilize NaHCO3 as an 8% solution and vitamin solution while gassing with N2. Aseptically and anaerobically add yeast extract, peptone, NaHCO3 and vitamin solutions, and sulfur to the medium under a N2-CO2 (80:20) atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O solution, and adjust pH to 6.2-6.4 with sterile 1 N H2SO4.


## FeCl2 solution

10	ml	HCl (7.7 M, 25%)	
1.5	g	FeCl2 x 4 H2O	
990	ml	Distilled water	


## Trace element solution

70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	


## 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