Desulfurella II Medium (DSMZ Medium 480c)

[所属分类:培养基配方] [发布时间:2020-6-11] [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

Desulfurella II Medium
(DSMZ Medium 480c)

培养基配方
Composition per liter:
MOPS [3-(N-morpholino) propane sulfonic acid].....................................3.0g
Sulfur, powder....................................................................1.0g
NH4Cl ...........................................................................0.33g
CaCl2 ·2H2O.....................................................................0.33g
MgCl2 ·6H2O.....................................................................0.33g
KCl..............................................................................0.33g
KH2PO4 ..........................................................................0.33g
Yeast extract.....................................................................0.1g
Resazurin .......................................................................1.0mg
NaHCO3 solution.................................................................40.0mL
Na2S·9H2O solution.............................................................10.0mL
Vitamin solution................................................................10.0mL
Substrate solution..............................................................10.0mL
Trace elements solution SL-10 ...................................................1.0mL
pH 6.9 ± 0.2 at 25°C
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 ...........................................................................2.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared
freshly.
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 compo-
nents. 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.
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. Sparge with N2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Substrate Solution:
Composition per 10.0mL:
Na-pyruvate .....................................................................2.5g
Preparation of Substrate Solution:  Add Na-pyruvate to distilled/
deionized water and bring volume to 10.0mL. Sparge with N2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store anaer-
obically.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free 80% N2 + 20% CO2  gas mixture. Add components, except
sulfur, substrate solution, vitamin solution, NaHCO3 solution, and
Na2S·9H2O solution, to 930.0mL distilled/deionized water. Mix thor-
oughly. Sparge for 30 min with 80% N2 + 20% CO2 . Adjust pH to 5.9
with concentrated NaOH. Distribute under 80% N2 + 20% CO2 into an-
aerobic tubes or bottles containing sulfur powder (100mg S per
10.0mL medium). Autoclave for 20 min at 110°C. Sparge with 80% N2
+ 20% CO2 . Aseptically and anaerobically add the appropriate
amounts to achieve concentrations of 10.0mL sterile vitamin solution,
10.0mL substrate solution, 10.0mL Na2S·9H2O solution, and 40.0mL
sterile NaHCO3 solution per liter of medium.
Use:  For the cultivation of Desulfurella propionica DSM 10410.