Methermicoccus Medium (DSMZ Medium 1084)

[所属分类:培养基配方] [发布时间:2021-5-27] [发布人:网站管理员2] [阅读次数:] [返回]

Methermicoccus Medium
(DSMZ Medium 1084)

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

Composition per liter:

NaCl ...........................................................................................24.0g
MgCl 2 ·6H 2 O................................................................................10.2g
Yeast extract .................................................................................2.0g
KCl .............................................................................................0.34g
NH 4 Cl ........................................................................................0.25g
K 2 HPO 4 ........................................................................................0.2g
Resazurin ..................................................................................0.5mg
Cysteine solution......................................................................10.0mL
Sulfide solution........................................................................10.0mL
Bicarbonate solution................................................................10.0mL
Coenzyme M solution..............................................................10.0mL
Methanol ...................................................................................8.0mL
Sludge fluid................................................................................5.0mL
pH 9.5 ± 0.2 at 25°C
Methanol:
Composition per 10.0mL:
Methanol .................................................................................10.0mL
Preparation of Methanol: Sparge 10.0mL of methanol with 100%
N 2 . Filter sterilize.
Coenzyme M Solution:
Composition per 10.0mL:
Mercaptoethanesulfonic acid (coenzyme M) ..............................2.5g
Preparation of Coenzyme M: Add coenzyme M to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
B icarbonate Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of Bicarbonate Solution:  Add NaHCO 3  to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO 2 + 80% H 2 . Filter sterilize.
Sulfide Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Sulfide Solution: Add Na 2 S·9H 2 O to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave
under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room
temperature.
Cysteine Solution:
Composition per 10.0mL:
L- Cysteine-HCl·2H 2 O ...................................................................0.3g
Preparation of Cysteine Solution: Add  L- cysteine-HCl·2H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Filter sterilize.
Sludge Fluid:
Composition per 500.0mL:
Yeast extract..................................................................................2.0g
Sludge ....................................................................................500.0mL
Preparation of Sludge Fluid:  Add yeast extract to sludge from an
anaerobic digester. Gas with nitrogen gas for a few minutes. Incubate
at 37°C for 24 hr. Centrifuge the sludge at 13,000 x g. Autoclave for 15
min at 15 psi pressure–121°C. Place the resulting, clear supernatant in
screw-capped vessels under nitrogen gas. The sludge fluid can be
stored at room temperature in the dark.
Preparation of Medium:  Add components, except bicarbonate,
sludge fluid, methanol, coenzyme M, cysteine, and sulfide solutions, to
distilled/deionized water and bring volume to 947.0mL. Gently heat
and bring to boiling. Boil medium for 1 min. Cool to room temperature
under 80% N 2  + 20% CO 2  gas mixture. Dispense under same gas atmo-
sphere into culture vessels. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature under 80% N 2  + 20% CO 2  gas mix-
ture. Aseptically add coenzyme M, methanol, sludge fluid, cysteine,
and sulfide from sterile anoxic stock solutions. Adjust pH to 6.0–6.5.
Use: For the cultivation of Methermicoccus spp.