Desulfomusa hansenii Medium (DSMZ Medium 916)

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfomusa hansenii Medium
(DSMZ Medium 916)

培养基配方
Solution A.................................................................870.0mL
Solution D..................................................................50.0mL
Solution F..................................................................50.0mL
Solution E (Vitamin solution)...............................................10.0mL
Solution G..................................................................10.0mL
Solution H..................................................................10.0mL
Solution B (Trace elements solution SL-10)...................................1.0mL
Solution C (Selenite-tungstate solution).....................................1.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per 870.0mL:
NaCl.........................................................................20.0g
Na2SO4 ......................................................................1.42g
KCl..........................................................................0.67g
NH4Cl ........................................................................0.1g
KH2PO4 ......................................................................0.01g
MgSO4 ·7H2O.................................................................0.02g
Resazurin ...................................................................0.5mg
Preparation of Solution A: Add components to 870.0mL of dis-
tilled/deionized water. Mix thoroughly. Gently heat and bring to boiling.
Boil for 3 min. Cool to room temperature while sparging with 80% N2  +
20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room
temperature.
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 (Selenite-Tungstate Solution):
Composition per liter:
NaOH...................................................................0.5g
Na2WO4 ·2H2O.........................................................4.0mg
Na2SeO3 ·5H2O........................................................3.0mg
Preparation of Solution C (Selenite-Tungstate Solution):
Add components to distilled/deionized water and bring volume to
1.0L. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution D:
Composition per 50.0mL:
NaHCO3 .............................................................5.0g
Preparation of Solution D:  Add NaHCO3 to distilled/deionized
water and bring volume to 50.0mL. 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 ........................................................0.1mg
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 . Filter sterilize.
Solution F:
Composition per 50.0mL:
MgCl2 ·6H2O ..........................................................10.6g
CaCl2 ·2H2O ..........................................................1.52g
Preparation of Solution F:  Add components to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution G:
Composition per 10.0mL:
FeCl2 ·4H2O...........................................................0.52g
Preparation of Solution G:  Add FeCl2 ·4H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to 2.0.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution H:
Composition per 10.0mL:
Na-propionate...........................................................0.96g
Preparation of Solution H:  Add Na-propionate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically under 80%
N2  + 20% CO2  sequentially add to 860.0mL sterile solution A, 1.0mL
solution B, 1.0mL solution C, 50.0mL solution D, 10.0mL solution E,
50.0mL solution F, 10.0mL solution G, and 10.0mL solution H. Asep-
tically and anaerobically distribute under 80% N2  + 20% CO2 into ap-
propriate vessels. The final pH should be 7.2. Addition of 10–20mg
sodium dithionite per liter from a 5% (w/v) solution, freshly prepared
under N2  and filter sterilized, may stimulate growth.
Use: For the cultivation of Desulfomusa hansenii.