Denitrovibrio Medium (DSMZ Medium 881)

[所属分类:培养基配方] [发布时间:2020-6-2] [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

Denitrovibrio Medium
(DSMZ Medium 881)

培养基配方
Composition per 1032.0mL:
NaCl.........................................................................20.0g
MgCl2 ·6H2O..................................................................3.0g
KH2PO4 .......................................................................1.0g
NaNO3 ........................................................................0.7g
KCl...........................................................................0.5g
NH4Cl .......................................................................0.25g
CaCl2 ·2H2O.................................................................0.15g
Na2SO4 ......................................................................0.02g
Resazurin ...................................................................0.5mg
Na-acetate solution ........................................................10.0mL
NaHCO3  solution ...........................................................10.0mL
Na2S·9H2O solution.........................................................10.0mL
Seven vitamin solution.......................................................1.0mL
Trace elements solution SL-10 ...............................................1.0mL
pH 6.8–7.2 at 25°C
Na-Acetate Solution:
Composition per 10.0mL:
Na-acetate ...................................................................1.64g
Preparation of Na-Acetate Solution: Add Na-acetate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O.....................................................................0.5g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 .......................................................................2.5g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ...................................................300.0mg
Thiamine-HCl·2H2O..........................................................200.0mg
Nicotinic acid..............................................................200.0mg
Vitamin B 12 ...............................................................100.0mg
Calcium pantothenate .......................................................100.0mg
p-Aminobenzoic acid..........................................................80.0mg
D (+)-Biotin.................................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O.................................................................1.5g
CoCl2 ·6H2O..............................................................190.0mg
MnCl2 ·4H2O .............................................................100.0mg
ZnCl2 .....................................................................70.0mg
Na2MoO4 ·2H2O.............................................................36.0mg
NiCl2 ·6H2O...............................................................24.0mg
H3BO3 ......................................................................6.0mg
CuCl2 ·2H2O................................................................2.0mg
HCl (25% solution).........................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 100%
N2 gas atmosphere. Add components, except NaHCO3  solution, Na-ac-
etate solution, Na2S·9H2O solution, seven vitamin solution, and trace
elements solution SL-10, to distilled/deionized water and bring volume
to 949.0mL. Mix thoroughly. Adjust pH to 6.8–7.2. Sparge with 80% N2
+ 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 10.0mL NaHCO3  solution, 10.0mL Na-acetate
solution, 10.0mL Na2S·9H2O solution, 1.0mL seven vitamin solution,
and 1.0mL trace elements solution SL-10. Mix thoroughly. Aseptically
and anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Denitrovibrio acetiphilus.