Acetobacteroides glycinophilus Medium

[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

Acetobacteroides glycinophilus Medium
Composition per 1020.0mL:
Na 2 HPO 4 .......................................................................................5.8g
KH 2 PO 4 .........................................................................................3.0g
NH 4 Cl ...........................................................................................1.0g
MgCl 2 ·6H 2 O .................................................................................0.2g
Resazurin ...................................................................................1.0mg
CaCl 2 ·2H 2 O ................................................................................0.13g
Trace elements solution ...........................................................10.0mL
Vitamin solution.........................................................................5.0mL
Yeast extract solution.................................................................5.0mL
Glycine solution.........................................................................5.0mL
NaHCO 3 solution.......................................................................5.0mL
Na 2 S·9H 2 O solution...................................................................5.0mL
pH 7.2–7.4 at 25°C
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.......................................................................2.8g
NaCl..............................................................................................1.0g
FeCl 3 ·4H 2 O...................................................................................0.2g
CoCl 2 ·6H 2 O ................................................................................0.17g
CaCl 2 ·2H 2 O...................................................................................0.1g
MnCl 2 ·4H 2 O..................................................................................0.1g
ZnCl 2 .............................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.026g
CuCl 2 ...........................................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO·2H 2 O............................................................................0.01g
Na 2 SeO·5H 2 O.............................................................................0.02g
Preparation of Trace Elements Solution : Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add distilled/deionized water to 1.0L. Add remain-
ing components. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................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.
Yeast Extract Solution:
Composition per 5.0mL:
Yeast extract..................................................................................0.5g
Preparation of Yeast Extract Solution : Add yeast extract to dis-
tilled/deionized water and bring volume to 5.0mL. Mix thoroughly.
Sparge under 100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N 2 gas.
Glycine Solution:
Composition per 5.0mL:
Glycine..........................................................................................1.5g
Preparation of Glycine Solution : Add glycine to distilled/deion-
ized water and bring volume to 5.0mL. Mix thoroughly. Sparge under
100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi pressure–121°C.
Store under N 2 gas.
NaHCO 3 Solution:
Composition per 5.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of NaHCO 3 Solution : Add NaHCO 3 to distilled/de-
ionized water and bring volume to 5.0mL. Mix thoroughly. Sparge un-
der 100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi pressure–
121°C. Store under N 2 gas.
Na 2 S·9H 2 O Solution:
Composition per 5.0mL:
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 5.0mL. Mix thoroughly.
Sparge under 100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N 2 gas.
Preparation of Medium: Add components, except yeast extract so-
lution, glycine solution, NaHCO 3 solution, and Na 2 S·9H 2 O solution, to
distilled/deionized water and bring volume to 1.0L. Adjust pH to 7.2–
7.4 with NaOH. Mix thoroughly. Gently heat and bring to boiling. Boil
for a few minutes. Allow to cool to room temperature under 100% N 2 .
Distribute into tubes or flasks under 100% N 2 . Autoclave for 15 min at
15 psi pressure–121°C. Cool to room temperature. Before inoculation,
aseptically and anaerobically add yeast extract solution, glycine solu-
tion, NaHCO 3 solution, and Na 2 S·9H 2 O solution.
Use:  For the cultivation and maintenance of Acetobacteroides glyci-
nophilus.