Acetobacterium 2 Medium

[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 2 Medium
Composition per 1050.0mL:
NaCl............................................................................................20.0g
NaHCO 3 ......................................................................................10.0g
Yeast extract..................................................................................2.0g
Ethylene glycol...........................................................................1.25g
NH 4 Cl ...........................................................................................1.0g
K 2 HPO 4 ......................................................................................0.45g
KH 2 PO 4 ......................................................................................0.33g
MgSO 4 ·7H 2 O................................................................................0.1g
Resazurin ...................................................................................1.0mg
Ethylene glycol solution ..........................................................20.0mL
Trace elements solution ...........................................................20.0mL
Vitamin solution.......................................................................20.0mL
Reducing agent solution ..........................................................10.0mL
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
CaCl 2 ·2H 2 O..................................................................................1.0g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
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 Trace Elements 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. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Reducing Agent Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O.....................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Reducing Agent Solution : Add 10.0mL of dis-
tilled/deionized water to a test tube. Gently heat and bring to boiling.
Boil under N 2 gas for 1 min. Cool to room temperature. Add  L -
cysteine·HCl·H 2 O and dissolve. Adjust pH to 9 with 5N NaOH. Add
washed Na 2 S·9H 2 O and dissolve. Autoclave for 10 min at 15 psi pres-
sure–121°C.
Ethylene Glycol Solution:
Composition per 20.0mL:
Ethylene glycol.............................................................................5.0g
Preparation of Ethylene Glycol Solution:  Add ethylene glycol
to distilled/deionized water and bring volume to 20.0mL. Mix thor-
oughly. Sparge under 100% N 2 gas for 3 min. Filter sterilize. Store un-
der N 2 gas.
Preparation of Medium: Add components, except NaHCO 3 , eth-
ylene glycol solution, and reducing agent solution, to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly. Gently heat and
bring to boiling. Boil for a few minutes. Allow to cool to room temper-
ature under 80% N 2 + 20% CO 2 . Add the NaHCO 3 and adjust pH to
7.4. Distribute into tubes or flasks under 80% N 2 + 20% CO 2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Before inoculation, add sterile anaerobic Na 2 CO 3 (0.25mL of 5%
Na 2 CO 3 per 10.0mL of medium) to bring the pH to 8.2. Add sterile eth-
ylene glycol solution and reducing agent solution.
Use:  For the cultivation and maintenance of Acetobacterium woodii.