Papillibacter Medium (DSMZ Medium 872)

[所属分类:培养基配方] [发布时间:2021-6-30] [发布人:网站管理员2] [阅读次数:] [返回]

Papillibacter Medium
(DSMZ Medium 872)

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

Composition per 1070.0mL:

Yeast extract..................................................................................5.0g
Trypticase™..................................................................................5.0g
NaCl..............................................................................................1.0g
Cysteine-HCl·H 2 O........................................................................0.5g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
NH 4 Cl ...........................................................................................0.3g
K 2 HPO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ..................................................................................0.1g
Resazurin ...................................................................................0.5mg
NaHCO 3 solution.....................................................................50.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Cinnamate solution..................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.0–7.5 at 25°C
Cinnamate Solution:
Composition per 10.0mL:
Na-trans-cinnamate.......................................................................0.8g
Preparation of Cinnamate Solution:  Add Na-trans-cinnamate to
distilled/deionized water and bring volume to 10.0mL. Sparge with N 2 .
Filter sterilie.
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 dis-
tilled/deionized water and bring volume to 10.0mL. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
NaHCO 3 Solution:
Composition per 100.0mL:
NaHCO 3 ......................................................................................10.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be
prepared freshly.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
H 3 BO 3 ....................................................................................300.0mg
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
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)....................................................................7.7mL
Preparation of Trace Elements Solution SL-10: Add 1.5g of
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 compo-
nents. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at
15 psi pressure–121°C. Cool to room temperature.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except cysteine,
NaHCO 3 solution, Na 2 S·9H 2 O solution, and cinnamate solution, to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently
heat and bring to boiling. Boil for 10 min. Cool to room temperature
while sparging with 80% N 2 + 20% CO 2 . Add 0.5g cysteine-HCl·H 2 O.
Mix thoroughly. Adjust pH to 7.0. Distribute into anaerobe tubes. Au-
toclave for 15 min at 15 psi pressure–121°C. For every 10.0mL medium
inject 0.5mL sterile NaHCO 3 solution, 0.1mL Na 2 S·9H 2 O solution, and
0.1mL cinnamate solution. Mix thoroughly. Final pH is 7.0–7.5.
Use:  For the cultivation of Papillibacter cinnamivorans.