Desulfatirhabdium Medium (DSMZ Medium 1086)

[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

Desulfatirhabdium Medium
(DSMZ Medium 1086)

培养基配方
Composition per liter:
Na2SO4 ......................................................................2.8g
Na2HPO4 ....................................................................0.53g
KH2PO4 .....................................................................0.41g
NH4Cl .......................................................................0.3g
NaCl ........................................................................0.3g
CaCl2 ·2H2O................................................................0.11g
MgCl2 ·6H2O.................................................................0.1g
Yeast extract ..............................................................0.02g
Crotonate solution ........................................................10.0mL
Benzoate solution .........................................................10.0mL
Vitamin solution ..........................................................10.0mL
Na2S·9H2O solution........................................................10.0mL
NaHCO3  solution ..........................................................10.0mL
Trace elements solution SL-10 ..............................................1.0mL
Selenite/tungstate solution ................................................1.0mL
pH 7.1 ± 0.2 at 25°C
NaHCO3 Solution :
Composition per 10.0mL:
NaHCO3 .....................................................................4.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
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% H 2 + 20% CO2 . Filter sterilize.
Crotonate Solution:
Composition per 10.0mL:
Na-crotonate ...............................................................1.7g
Preparation of Crotonate Solution:  Add components 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.
Benzoate Solution:
Composition per 10.0mL:
Na-benzoate ................................................................0.43g
Preparation of Benzoate Solution:  Add components 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.
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.
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.
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 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, except bicarbonate, vi-
tamins, crotonate, benzoate, and sulfide solutions, to distilled/deion-
ized water and bring volume to 950.0mL. Mix thoroughly. Gently heat
and bring to boiling. Boil for 1 min. Cool while sparging with 80% N2
+ 20% CO2 . Dispense  under  80% N2 + 20% CO2 into culture vessels.
Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anox-
ically add vitamins, crotonate, benzoate, and sulfide. Adjust the final
pH of the medium to 7.0–7.2. After inoculation pressurize the vessels
with 80% N2 + 20% CO2 to 0.7 bar overpressure.
Use: For the cultivation of Desulfatirhabdium spp.