Desulfurella Medium

[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 Medium

培养基配方
Composition per liter:
Sulfur, powdered.............................................................10.0g
Sodium acetate................................................................5.0g
CaCl2 ·2H2O.................................................................0.33g
KCl..........................................................................0.33g
KH2PO4 ......................................................................0.33g
MgCl2 ·6H2O ................................................................0.33g
NH4Cl .......................................................................0.33g
Yeast extract.................................................................0.1g
Resazurin ..................................................................1.0 mg
NaHCO3  solution.............................................................40.0mL
Na2S·9H2O solution..........................................................10.0mL
Wolfe's vitamin solution ....................................................10.0mL
Trace elements solution SL-10 ...............................................1.0 mL
pH 6.8–7.0 at 25°C
NaHCO3 Solution:
Composition per 40.0mL:
NaHCO3 ........................................................................2.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 40.0mL. Mix thoroughly. Gas under
100% N2 . Autoclave for 15 min at 15 psi pressure–121°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.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Wolfe's Vitamin Solution:
Composition per liter:
Pyridoxine·HCl................................................................10.0mg
Calcium  D -(+)-pantothenate....................................................5.0mg
Nicotinic acid..................................................................5.0mg
p-Aminobenzoic acid.............................................................5.0mg
Riboflavin .....................................................................5.0mg
Thiamine·HCl ..................................................................5.0mg
Thioctic acid ..................................................................5.0mg
Biotin .........................................................................2.0mg
Folic acid......................................................................2.0mg
Cyanocobalamine..............................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. 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 compo-
nents. Mix thoroughly. Sparge with 100% N2 . Autoclave for 15 min at
15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 . Add components, except NaHCO3  solution, Wolfe's vi-
tamin solution, and Na2S·9H2O solution, to distilled/deionized water
and bring volume to 940.0mL. Mix thoroughly. Adjust pH to 5.9. Do
not autoclave. Sterilize medium by heating to 100°C for 1 hr on 3 con-
secutive days. Prior to inoculation, aseptically and anaerobically add
40.0mL of sterile NaHCO3  solution, 10.0mL of sterile Wolfe's vitamin
solution, and 10.0mL of sterile Na2S·9H2O solution. Mix thoroughly.
Final pH should be 6.8–7.0.
Use:  For the cultivation of Desulfurella species, especially Desul-
furella acetivorans.