Acidolobus aceticus Medium (DSMZ Medium 901)

[所属分类:培养基配方] [发布时间:2019-9-23] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Acidolobus aceticus Medium
(DSMZ Medium 901)
Composition per 1055mL:
Sulfur, powdered.........................................................................10.0g
NH 4 Cl .........................................................................................0.33g
KCl..............................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
MgCl 2 ·6H 2 O ...............................................................................0.33g
CaCl 2 ·2H 2 O ................................................................................0.33g
Resazurin ...................................................................................0.5mg
Yeast extract solution...............................................................30.0mL
Na 2 S·9H 2 O solution.................................................................15.0mL
Vitamin solution.......................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 3.5–3.8 at 25°C
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 . 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.
Na 2 S·9H 2 O Solution :
Composition per 20.0mL:
Na 2 S·9H 2 O....................................................................................0.6g
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 20.0mL. Mix thoroughly.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature. Neturalize to pH 7.0 with HCl.
Yeast Extract Solution:
Composition per 30.0mL:
Yeast extract..................................................................................3.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 30.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave under 100% N 2 for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Preparation of Medium: Prepare and dispense medium under
100% CO 2 . Add components, except vitamin solution, Na 2 S·9H 2 O so-
lution, and yeast extract solution, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Adjust pH to 3.5 with H 2 SO 4 . Distrib-
ute to anaerobe tubes or bottles. Heat to 90°C for 1 hr on each of 3 suc-
cessive days. Aseptically and anaerobically add, per liter of medium,
10.0mL sterile vitamin solution, 15.0mL of sterile Na 2 S·9H 2 O solu-
tion, and 30.0mL sterile yeast extract solution. Mix thoroughly. The fi-
nal pH should be 3.5–3.8.
Use:  For the cultivation of Acidilobus aceticus.