FUF Medium (DSMZ Medium 318a)

[所属分类:培养基配方] [发布时间:2021-3-4] [发布人: ] [阅读次数:] [返回]

FUF Medium

(DSMZ Medium 318a)

培养基配方

Composition per liter:
KHCO 3 ..........................................................................................2.0g
NH 4 Cl ...........................................................................................1.0g
NaCl..............................................................................................0.6g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O .................................................................................0.1g
CaCl 2 ·2H 2 O ................................................................................0.08g
Resazurin...................................................................................1.0mg
HEPES solution.......................................................................50.0mL
Furoic acid solution .................................................................50.0mL
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Cysteine solution .....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.8 ± 0.2 at 25°C
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................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% H 2 + 20% CO 2 . Filter sterilize.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................0.3g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to distilled/deionized 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.
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................12.8g
FeCl 3 ·6H 2 O.................................................................................1.35g
NaCl..............................................................................................1.0g
NiCl 2 ·6H 2 O.................................................................................0.12g
MgCl 2 ·4H 2 O..................................................................................0.1g
CaCl 2 ·2H 2 O...................................................................................0.1g
ZnCl 2 .............................................................................................0.1g
Na 2 SeO 3 ·5H 2 O..........................................................................0.026g
CuCl 2 ·2H 2 O ..............................................................................0.025g
CoCl 2 ·6H 2 O ..............................................................................0.024g
Na 2 MoO 4 ·4H 2 O ........................................................................0.024g
H 3 BO 3 .........................................................................................0.01g
Preparation of Trace Elements Solution: Add nitrilotriacetic acid to 200.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Mix thoroughly.
Furoic Acid Solution:
Composition per 100.0mL:
2-Furoic acid.................................................................................4.4g
Preparation of Furoic Acid Solution: Add furoic acid to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Adjust pH to 7.0 with NaOH. Sparge with 100% N 2 . Filter sterilize.
HEPES Solution:
Composition per 100.0mL:
HEPES..........................................................................................6.2g
Preparation of HEPES Solution: Add HEPES to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except cysteine solution, furoic acid solution, Na 2 S·9H 2 O solution, and vitamin solution, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL cysteine solution, 50.0mL furoic acid solution, 10.0mL Na 2 S·9H 2 O solution, and 10.0mL vitamin solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles. After inoculation, flush and repressurize the gas head space of culture bottles with sterile 80% N 2 + 20% CO 2 to 1 bar overpressure.
Use:  For the cultivation of Methanosarcina thermophila (Methanosarcina sp.) and Methanosarcina mazei=Methanococcus mazei (Methanosarcina frisia).