Desulfonatronospira Medium (DSMZ Medium 1101)

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfonatronospira Medium
(DSMZ Medium 1101)

培养基配方
Composition per liter:
Na2CO3  ...................................................................95.0g
NaHCO3  ...................................................................15.0g
NaCl.......................................................................15.0g
K2HPO4 .....................................................................0.5g
Ammonium chloride solution................................................10.0mL
Magnesium chloride solution...............................................10.0mL
Yeast extract solution....................................................10.0mL
Lactate solution..........................................................10.0mL
Sulfite solution..........................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Vitamin solution .........................................................10.0mL
Trace elements solution SL-10 with EDTA....................................1.0mL
Selenite/tungstate solution ...............................................1.0mL
pH 10.0 ± 0.2 at 25°C
Trace Elements Solution SL-10 with EDTA:
Composition per liter:
FeCl2 ·4H2O................................................................1.5g
Na2 -EDTA...................................................................0.5g
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 with EDTA:
Add FeCl2 ·4H2O 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. Sparge with 100% N2 . Autoclave for 15 min
at 15 psi pressure–121°C. Adjsut pH to 7.0.
Selenite/Tungstate Solution:
Composition per liter:
NaOH.......................................................................0.5g
Na2WO4 ·2H2O.............................................................4.0mg
Na2SeO3 ·5H2O............................................................3.0mg
Preparation of Selenite/Tungstate Solution:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl..........................................................10.0mg
Thiamine-HCl·2H2O.......................................................5.0mg
Riboflavin ..............................................................5.0mg
Nicotinic acid...........................................................5.0mg
D -Ca-pantothenate.......................................................5.0mg
p-Aminobenzoic acid......................................................5.0mg
Lipoic acid .............................................................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. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Lactate Solution:
Composition per 10.0mL:
Na-lactate.................................................................2.2g
Preparation of Lactate Solution:  Add Na-lactate to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Sulfite Solution:
Composition per 10.0mL:
Na2SO3 .....................................................................1.3g
Preparation of Sulfite:  Add Na2SO3 to distilled/deionized water
and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 .
Filter sterilize.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O...................................................................0.24g
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.
Ammonium Chloride Solution:
Composition per 10.0mL:
NH4Cl ........................................................................0.2g
Preparation of Ammonium Chloride Solution:  Add NH4Cl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Magnesium Chloride Solution:
Composition per 10.0mL:
MgCl2 ·6H2O .................................................................0.2g
Preparation of Magnesium Chloride Solution:  Add MgCl2 ·6H2O
to distilled/deionized water and bring volume to 10.0mL. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract ..................................................................0.5g
Preparation of Yeast Extract Solution:  Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Pyruvate Solution:
Composition per 10.0mL:
Na-pyruvate ...................................................................2.2g
Preparation of Pyruvate Solution:  Add Na-pyruvate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Thiosulfate Solution:
Composition per 10.0mL:
Na2S2O3 .......................................................................5.0g
Preparation of Thiosulfate Solution:  Add Na2S2O3  to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
Preparation of Medium:  Add components, except ammonium
chloride, magnesium chloride, yeast extract, trace elements, vitamin,
pyruvate, thiosulfate, and sulfide solutions, to distilled/deionized water
and bring volume to 930.0mL. Gently heat and bring to boiling. Boil
for 1 min. Mix thoroughly. Cool to room temperature while sparging
with 100% N2 . Dispense into culture vessels under an atmosphere of
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
add the ammonium chloride, magnesium chloride, yeast extract, trace
elements, vitamin, pyruvate, thiosulfate, and sulfide solutions. Adjust
the pH to 10.
Use: For the cultivation of Desulfonatronospira delicata.