Dethiobacter Medium (DSMZ Medium 1104)

[所属分类:培养基配方] [发布时间:2020-6-12] [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

Dethiobacter Medium
(DSMZ Medium 1104)

培养基配方
Composition per liter:
Na2CO3  ...................................................................22.0g
NaHCO3  ....................................................................8.0g
NaCl........................................................................6.0g
Sulfur, powdered............................................................1.5g
K2HPO4 .....................................................................0.5g
Thiosulfate solution......................................................10.0mL
Vitamin solution .........................................................10.0mL
Ammonium chloride solution................................................10.0mL
Magnesium chloride solution ..............................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Yeast extract solution ...................................................10.0mL
Acetate solution..........................................................10.0mL
Butyrate solution.........................................................10.0mL
Trace element ssolution SL-10 with EDTA....................................1.0mL
Selenite/tungstate solution ...............................................1.0mL
pH 9.5 ± 0.2 at 25°C
Ammonium Chloride Solution:
Composition per 10.0mL:
NH4Cl ......................................................................0.2g
Preparation of Ammonium Chloride Solution:  Add NH 4 Cl 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.
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.
Thiosulfate Solution:
Composition per 10.0mL:
Na2S2O3 ....................................................................2.5g
Preparation of Thiosulfate Solution:  Add Na2S2O3 to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
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.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract ...........................................................0.05g
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.
Acetate Solution:
Composition per 10.0mL:
Sodium acetate............................................................0.4g
Preparation of Acetate Solution:  Add sodium acetate 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.
Butyrate Solution:
Composition per 10.0mL:
Sodium butyrate...........................................................1.1g
Preparation of Butyrate Solution:  Add sodium butyrate 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.
Preparation of Medium:  Add components, except ammonium
chloride, magnesium chloride, yeast extract, trace elements, vitamin,
ethanol, acetate, sulfur, thiosulfate, and sulfide solutions, to distilled/
deionized water and bring volume to 920.0mL. 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, ethanol, acetate, sulfur, thiosul-
fate, and sulfide solutions. Sulfur is sterilized by steaming for 3 h on
each of 3 successive days and added as powder. Adjust pH of final me-
dium to 9.5. After inoculation add sterile H2 gas to 0.5 bar overpres-
sure.
Use: For the cultivation of Desulfurivibrio alkaliphilus.