Moorella glycerini Medium (DSMZ Medium 793)

[所属分类:培养基配方] [发布时间:2021-6-7] [发布人:网站管理员2] [阅读次数:] [返回]

Moorella glycerini Medium
(DSMZ Medium 793)

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

Composition per liter:

NaHCO 3 ......................................................................................10.0g
Yeast extract..................................................................................0.5g
KH 2 PO 4 .......................................................................................0.33g
NH 4 Cl .........................................................................................0.33g
KCl..............................................................................................0.33g
MgCl 2 ·6H 2 O................................................................................0.33g
CaCl 2 ·2H 2 O.................................................................................0.33g
Resazurin ...................................................................................0.5mg
Vitamin solution.......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Glycerol, 87%............................................................................3.0mL
Trace elements solution SL-10 ..................................................1.0mL
Selenite-tungstate solution.........................................................1.0mL
pH 6.7± 0.2 at 25°C
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/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.
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.
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.
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 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Prepare and dispense medium under 100%
CO 2 gas atmosphere. Add components, except NaHCO 3 , Na 2 S·9H 2 O
solution, and vitamin solution, to distilled/deionized water and bring
volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling.
Boil for 5 min. Cool to room temperature while sparging with 100%
CO 2 . Add solid NaHCO 3 . Adjust pH to 6.7 by adding NaOH and equil-
ibrating with 100% CO 2 . Distribute into tubes or bottles. Autoclave for
20 min at 15 psi pressure–121°C. Aseptically and anaerobically add
10.0mL Na 2 S·9H 2 O solution and vitamin solution, 0.1mL per 10mL
medium. Mix thoroughly. Aseptically and anaerobically distribute into
sterile tubes or bottles.
Use:  For the cultivation of Moorella glycerini.