Methanobrevibacter curvatus Medium (DSMZ Medium 734)

[所属分类:培养基配方] [发布时间:2021-5-24] [发布人:网站管理员2] [阅读次数:] [返回]

Methanobrevibacter curvatus Medium
(DSMZ Medium 734)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1010.0mL:

NaCl..............................................................................................1.0g
KCl............................................................................................... 0.5g
Casamino acids.............................................................................0.5g
Yeast extract..................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
NH 4 Cl ...........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.2g
Na 2 SO 4 ........................................................................................0.15g
CaCl 2 ·2H 2 O ..................................................................................0.1g
Resazurin ...................................................................................0.5mg
NaHCO 3  solution.....................................................................40.0mL
Dithionite solution...................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
Selenite-tungstate solution.........................................................1.0mL
Seven vitamin solution ..............................................................1.0mL
pH 7.4 ± 0.2 at 25°C
NaHCO 3 Solution:
Composition per 40.0mL:
NaHCO 3 ........................................................................................5.8g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 40.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared
freshly.
Selenite-Tungstate Solution
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Selenite-Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Dithionite Solution
Composition per 10.0mL:
Na-dithionite..............................................................................2.0mg
Preparation of Dithionite Solution: Add Na-dithionite to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O
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% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................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.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solu-
tion, seven vitamin solution, selenite-tungstate solution, and trace ele-
ments solution SL-10, to distilled/deionized water and bring volume to
947.0mL. Mix thoroughly. Adjust pH to 7.6. Sparge with 80% H 2 + 20%
CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and an-
aerobically add 40.0mL NaHCO 3  solution, 1.0mL selenite-tungstate
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. Adjust pH to 7.6. Prior to inoculation add di-
thionite solution (0.1mL per 10mL medium) as reductant.
Use:  For the cultivation of Methanobrevibacter curvatus.