Anoxybacillus Medium (DSMZ Medium 898)

[所属分类:培养基配方] [发布时间:2019-10-21] [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

Anoxybacillus Medium
(DSMZ Medium 898)
Composition per liter:
NaHCO 3 ......................................................................................10.0g
NaCl..............................................................................................5.0g
Na 2 CO 3 .......................................................................................2.76g
NH 4 Cl ...........................................................................................1.0g
Yeast extract..................................................................................0.5g
KH 2 PO 4 .........................................................................................0.2g
KCl................................................................................................0.2g
MgCl 2 ·6H 2 O .................................................................................0.1g
Resazurin...................................................................................0.5mg
Glucose solution......................................................................50.0mL
Vitamin solution.......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 9.5–9.7 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.
Before use, neutralize to pH 7.0 with sterile HCl.
Glucose Solution:
Composition per 50.0mL:
Glucose .........................................................................................5.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. 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%
N 2 . Add components, except glucose solution, Na 2 S·9H 2 O solution,
and vitamin solution, to distilled/deionized water and bring volume to
930.0mL. Mix thoroughly. Sparge with 100% N 2  for 30–60 min. Adjust
pH to 8.0–8.5 with NaOH. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 50.0mL sterile glucose solu-
tion, 10.0mL sterile Na 2 S·9H 2 O solution, and 10.0mL sterile vitamin
solution. Mix thoroughly. The final pH should be 9.5–9.7. Aseptically
and anaerobically under 100% N 2  distribute into sterile tubes or bottles.
Use:  For the cultivation of Anoxybacillus pushchinoensis (Anoxybacil-
lus pushchinensis).