Acetobacterium 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 Medium
Composition per 1001.0mL:
Solution A..............................................................................870.0mL
Solution C..............................................................................100.0mL
Solution D................................................................................10.0mL
Solution E (Vitamin solution)..................................................10.0mL
Solution F.................................................................................10.0mL
Solution B (Trace elements solution SL-10) .............................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
Na 2 SO 4 ..........................................................................................3.0g
NaCl..............................................................................................1.0g
Yeast extract..................................................................................0.5g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O..................................................................................0.4g
NH 4 Cl ...........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
Resazurin ...................................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 3–4 min. Allow to cool to room
temperature while gassing under 80% N 2 + 20% CO 2 . Continue gas-
sing until pH reaches below 6.0. Seal the flask under 80% N 2 + 20%
CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution B (Trace Elements Solution SL-10 ):
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O ...........................................................................190.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Solution B (Trace Elements Solution SL-10):
Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Gas under 100% N 2 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of Solution C:  Add NaHCO 3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Gas under 80% N 2 + 20% CO 2 .
Solution D:
Composition per 10.0mL:
Methoxyacetate.............................................................................0.9g
Preparation of Solution D:  Add methoxyacetate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Gas under
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E (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 Solution E (Vitamin Solution): Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution F:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.4g
Preparation of Solution F:  Add Na 2 S·9H 2 O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically combine
solution A with solution B, solution C, solution D, solution E, and solu-
tion F, in that order. Mix thoroughly. Anaerobically distribute into ster-
ile tubes or flasks under 80% N 2 + 20% CO 2 .
Use:  For the cultivation and maintenance of Acetobacterium species.