Desulfovibrio aespoeensis Medium (DSMZ Medium 721)

[所属分类:培养基配方] [发布时间:2020-6-9] [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

Desulfovibrio aespoeensis Medium
(DSMZ Medium 721)

培养基配方
Composition per 1004.0mL:
Solution A................................................................870.0mL
Solution C................................................................100.0mL
Solution D.................................................................10.0mL
Solution E (Vitamin solution)..............................................10.0mL
Solution F.................................................................10.0mL
Selenite-tungstate solution.................................................2.0mL
Solution B (Trace elements solution SL-10)..................................1.0mL
Seven vitamin solution......................................................1.0mL
pH 7.3–7.5 at 25°C
Solution A:
Composition per 870.0mL:
NaCl.........................................................................7.0g
Na2SO4 ......................................................................3.0g
MgCl2 ·6H2O ................................................................1.3g
KCl..........................................................................0.5g
NH4Cl .......................................................................0.3g
KH2PO4 ......................................................................0.2g
CaCl2 ·2H2O ...............................................................0.15g
Resazurin ..................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly.
Solution B (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 Solution B (Trace Elements Solution SL-10):
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.
Solution C:
Composition per 100.0mL:
NaHCO3 .....................................................................5.0g
Preparation of Solution C: Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Flush with 80% N2  + 20% CO2 to remove dissolved oxygen.
Solution D:
Composition per 10.0mL:
Na-lactate..................................................................2.5g
Preparation of Solution D: Add Na-lactate 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.
Solution E (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 ..............................................................0.10mg
Preparation of Solution E (Vitamin Solution): Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution F:
Composition per 10.0mL:
Na2S·9H2O...................................................................0.4g
Preparation of Solution F:  Add Na2S·9H2O 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.
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% N 2 .
Mix thoroughly. Filter sterilize.
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 . Filter sterilize.
Preparation of Medium: Gently heat solution A and bring to boil-
ing. Boil solution A for a few minutes. Cool to room temperature. Gas
with 80% N2  + 20% CO2 gas mixture to reach a pH below 6. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se-
quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solu-
tion D, 10.0mL solution E, 10.0mL solution F, 2.0mL selenite tungstate
solution, and 1.0mL seven vitamin solution. Distribute anaerobically
under 80% N2  + 20% CO2 into appropriate vessels.
Use: For the cultivation of Desulfovibrio aespoeensis.