Acetobacterium carbinolicum 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

Acetobacterium carbinolicum Medium
Composition per 1011.2mL:
NaHCO 3 ........................................................................................4.5g
Na 2 SO 4 ........................................................................................2.84g
NaCl............................................................................................1.17g
Yeast extract..................................................................................1.0g
MgCl 2 ·6H 2 O .................................................................................0.4g
KCl................................................................................................0.3g
NH 4 Cl .........................................................................................0.27g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin...................................................................................0.5mg
Reducing agent solution ..........................................................10.0mL
Ethanol solution.........................................................................1.2mL
Trace elements solution.............................................................1.0mL
Vitamin solution.........................................................................1.0mL
pH 7.0–7.2 at 25°C
Trace Elements Solution:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................120.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................68.0mg
H 3 BO 3 ......................................................................................62.0mg
Na 2 MoO 4 ·2H 2 O...................................................................... 24.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
CuCl 2 ·2H 2 O.............................................................................17.0mg
HCl (0.05M solution)...........................................................1000.0mL
Preparation of Trace Elements Solution: Add components one
at a time to 1.0L of 0.05M HCl solution. Mix thoroughly.
Vitamin Solution:
Composition per 100.0mL
Thiamine·HCl..........................................................................10.0mg
p-Aminobenzoic acid.................................................................4.0mg
D (+)-Biotin.................................................................................1.0mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize. Flush with 80% N 2 + 20% CO 2 .
Reducing Agent Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
Preparation of Reducing Agent Solution:  Add Na 2 S·9H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave for 10 min at 15 psi pressure–121°C.
Ethanol Solution:
Composition per 10.0mL:
Ethanol (95% solution)............................................................10.0mL
Preparation of Ethanol Solution:  Filter sterilize. Sparge with N 2
gas for 1 min.
Preparation of Medium: Add components, except NaHCO 3 , etha-
nol, and reducing agent solution, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Boil
for a few minutes. Allow to cool to room temperature under 80% N 2 +
20% CO 2 . Add the NaHCO 3 and adjust pH to 6.9–7.1. Distribute into
tubes or flasks under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C. Cool to room temperature. Before inoculation,
add sterile anaerobic Na 2 CO 3 (0.25mL of 5% Na 2 CO 3 per 10.0mL of
medium) to bring the pH to 8.2. Add sterile ethanol and reducing agent
solution.
Use:  For the cultivation and maintenance of Acetobacterium malicum and
Acetobacterium carbinolicum.