Desulfuromonas acetexigenes Medium (DSMZ Medium 647)

[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

Desulfuromonas acetexigenes Medium
(DSMZ Medium 647)

培养基配方
Composition per liter:
NaCl...................................................................20.0g
Sulfur, powdered........................................................5.0g
MgCl2 ·6H2O............................................................2.0g
KH2PO4 .................................................................1.0g
MgSO4 ·7H2O............................................................1.0g
NH4Cl ..................................................................0.5g
CaCl2 ·2H2O ...........................................................0.1g
NaHCO3  solution......................................................50.0mL
Na-pyruvate solution .................................................10.0mL
Na2S·9H2O solution...................................................10.0mL
Seven vitamin solution ................................................1.0mL
Trace elements solution SL-10..........................................1.0mL
pH 7.2 ± 0.2 at 25°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.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 .................................................................5.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Na-pyruvate Solution:
Composition per 10.0mL:
Na-pyruvate ............................................................0.6g
Preparation of Na-pyruvate Solution: Add Na-pyruvate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride..............................................300.0mg
Thiamine-HCl·2H2O....................................................200.0mg
Nicotinic acid........................................................200.0mg
Vitamin B 12 .........................................................100.0mg
Calcium pantothenate .................................................100.0mg
p-Aminobenzoic acid....................................................80.0mg
D (+)-Biotin...........................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N2 .
Mix thoroughly. Filter sterilize.
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 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
Preparation of Medium: Sulfur is sterilized by steaming for 3 hr on
each of 3 successive days. Prepare and dispense medium under 80% N2
+ 20% CO2 gas atmosphere. Add components, except sulfur, NaHCO3
solution, Na-pyruvate solution, Na2S·9H2O solution, seven vitamin so-
lution, and trace elements solution SL-10, to distilled/deionized water
and bring volume to 928.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 5.0g sterile sulfur, 50.0mL
NaHCO3  solution, 10.0mL Na-pyruvate solution, 10.0mL Na2S·9H2O
solution, 1.0mL seven vitamin solution, and 1.0mL trace elements so-
lution SL-10. Mix thoroughly. Aseptically and anaerobically distribute
into sterile tubes or bottles.
Use:  For the cultivation of Desulfuromonas acetexigenes DSM8988.