Acetobacterium Medium (ATCC Medium 1019)

[所属分类:培养基配方] [发布时间:2019-9-18] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Acetobacterium Medium
(ATCC Medium 1019)
Composition per liter:
NaHCO 3 ........................................................................................3.0g
Yeast extract..................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
KH 2 PO 4 .........................................................................................0.4g
K 2 HPO 4 .........................................................................................0.4g
MgSO 4 ·7H 2 O................................................................................0.1g
Fructose (20% solution)...........................................................25.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Wolfe’s mineral solution..........................................................10.0mL
Resazurin (0.01% solution)........................................................1.0mL
pH 6.7 ± 0.2 at 25°C
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl ............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium pantothenate ................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Thioctic acid ..............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Cyanocobalamin .....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
MnSO 4 ·H 2 O..................................................................................0.5g
NaCl..............................................................................................1.0g
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O ..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add distilled/deionized water to 1.0L. Add remain-
ing components.
Preparation of Medium: Add all components, except fructose, to
distilled/deionized water and bring volume to 975.0mL. Boil to remove
dissolved O 2 . Add 40.0mL of a solution containing 1.25%  L-
cysteine·HCl·H 2 O and 1.25% Na 2 S·9H 2 O. Autoclave for 15 min at 15
psi pressure–121°C. Immediately gas with 90% N 2 + 10% CO 2 to
maintain anaerobiosis until cooled to 50°C. Add 25.0mL of a filter-
sterilized 20% fructose solution. If necessary, adjust pH to 6.7. Asepti-
cally distribute into tubes under anaerobic conditions. Cap with rubber
stoppers.
Use: For the cultivation and maintenance of Acetobacterium species.