Acetobacterium Medium (DSMZ 614)

[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
(DSMZ 614)
Composition per liter:
MgCl 2 ·6H 2 O ...............................................................................0.52g
Yeast extract..................................................................................0.5g
KCl..............................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
NH 4 Cl .........................................................................................0.33g
CaCl 2 ·2H 2 O ................................................................................0.22g
Resazurin ...................................................................................1.0mg
Fructose solution....................................................................100.0mL
Wolfe’s mineral solution..........................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
NaHCO 3  solution.....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.8–7.0 at 25°C
Fructose Solution:
Composition per 100.0mL:
D -Fructose...................................................................................10.0g
Preparation of Fructose Solution:  Add fructose to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
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
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoCl 2 ·6H 2 O ..................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.7g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Preparation of Medium: Prepare and dispense medium under
100% N 2 . Add components, except fructose solution, NaHCO 3 solution,
and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume
to 880.0mL. Mix thoroughly. Adjust pH to 6.8–7.0. Sparge with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and an-
aerobically add 100.0mL of sterile fructose solution, 10.0mL of sterile
NaHCO 3 solution, and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thor-
oughly. Aseptically and anaerobically distribute into sterile tubes or bot-
tles.
Use: For the cultivation of Acetobacterium bakii and Acetobacterium
paludosum.