ACB90 Medium (DSMZ Medium 298h)

[所属分类:培养基配方] [发布时间:2019-9-18] [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

ACB90 Medium
(DSMZ Medium 298h)
Composition per liter:
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
NH 4 Cl .........................................................................................0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin ...................................................................................1.0mg
NaHCO 3  solution.....................................................................10.0mL
Butanediol solution..................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Vitamin solution.......................................................................10.0mL
Seven vitamin solution ............................................................10.0mL
Glucose solution ......................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
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.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Filter sterilize.
Butanediol Solution:
Composition per 10.0mL:
2,3 butanediol ...............................................................................0.9g
Preparation of Butanediol Solution: Add butanediol to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin..................................................................................5.0mg
ACE Medium  19
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.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................200.0mg
Nicotinic acid.........................................................................200.0mg
Vitamin B 12 ............................................................................100.0mg
Calcium pantothenate ............................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
D (+)-Biotin...............................................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. Filter sterilize.
Cellobiose Solution:
Composition per 10.0mL:
Cellobiose .....................................................................................0.7g
Preparation of Cellobiose Solution: Add cellobiose to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. 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 NaHCO 3  solu-
tion, butanediol solution, Na 2 S·9H 2 O solution, vitamin solution, seven
vitamin solution, cellobiose solution, and trace elements solution SL-
10, to distilled/deionized water and bring volume to 939.0mL. Mix thor-
oughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave
for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add
10.0mL NaHCO 3  solution, 10.0mL butanediol solution, 10.0mL
Na 2 S·9H 2 O solution, 10.0mL vitamin solution, 10.0mL seven vitamin
solution, 10.0mL cellobiose solution, and 1.0mL trace elements solu-
tion SL-10. 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 an unclassified bacterium (Brevigemma cel-
lulytica) DSM 11249.