Desulfitobacterium PCE Medium (DSMZ Medium 717)

[所属分类:培养基配方] [发布时间:2020-6-4] [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

Desulfitobacterium PCE Medium
(DSMZ Medium 717)

培养基配方
Composition per liter:
(NH4)H2PO4 ..................................................................2.88g
MgSO4 ·7H2O..................................................................0.1g
Yeast extract.................................................................0.1g
Ca(NO3)2 ·4H2O .............................................................0.05g
Resazurin ...................................................................0.1mg
NaHCO3 solution.............................................................50.0mL
KOH solution................................................................20.0mL
Na-lactate solution.........................................................20.0mL
Na-fumarate solution........................................................20.0mL
Vitamin solution............................................................10.0mL
Na2S·9H2O solution..........................................................3.3mL
Seven vitamin solution ......................................................1.0mL
Trace elements solution SL-10................................................1.0mL
Selenite-tungstate solution..................................................1.0mL
pH 7.1 ± 0.2 at 25°C
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.
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.
KOH Solution:
Composition per 100.0mL:
KOH.........................................................................10.0g
Preparation of KOH Solution:  Add KOH to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . 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.
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.
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.
Na-lactate Solution:
Composition per 100.0mL:
Na-lactate.................................................................25.0g
Preparation of Na-lactate Solution: Add Na-lactate to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Na-fumarate Solution:
Composition per 100.0mL:
Na-fumarate................................................................16.0g
Preparation of Na-fumarate Solution: Add Na-fumarate to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Cool to room temperature.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, Na2S·9H2O solution, KOH solution, Na-lactate solution, Na-fu-
marate solution, vitamin solution, seven vitamin solution, selenite-
tungstate solution, and trace elements solution SL-10, to distilled/de-
ionized water and bring volume to 873.7mL. Mix thoroughly. Adjust pH
to 7.0–7.2. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C. Aseptically and anaerobically add 50.0mL
NaHCO3  solution, 3.3mL Na2S·9H2O solution, 20.0mL KOH solution,
20.0mL Na-lactate solution, 20.0mL Na-fumarate solution, 10.0mL vi-
tamin solution, 1.0mL seven vitamin solution, 1.0mL selenite-tung-
state solution, and 1.0mL trace elements solution SL-10. Mix
thoroughly. Aseptically and anaerobically distribute into sterile tubes or
bottles.
Use:  For the cultivation of Desulfitobacterium spp. and Desulfitobacte-
rium hafniense.