​Desulfurella II Medium (DSMZ Medium 480c)

[所属分类:培养基配方] [发布时间:2020-6-10] [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 components. Mix thor-
oughly. 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.
Substrate Solution:
Composition per 10.0mL:
Na-lactate......................................................................2.5g
Preparation of Substrate Solution:  Add Na-lactate 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 20 min at 110°C. Sparge with 80% N2 +
20% CO2 . Aseptically and anaerobically add 10.0mL sterile
Na2S·9H2O solution, 10.0mL sterile vitamin solution, 10.0mL sub-
strate solution, and 40.0mL sterile NaHCO3 solution per liter of medi-
um.
Use:  For the cultivation of Thermoproteus uzoniensis and Desulfurella
kamchatkensis DSM 10409.