Acetobacterium sp. Medium (DSMZ Medium 614)

[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 sp. Medium
(DSMZ Medium 614)
Composition per liter:
MgCl 2 ·6H 2 O ...............................................................................0.52g
Yeast extract..................................................................................0.5g
KCl..............................................................................................0.33g
NH 4 Cl .........................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
CaCl 2 ·2H 2 O ................................................................................0.22g
Resazurin...................................................................................1.0mg
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
NaHCO 3 solution.....................................................................10.0mL
Substrate solution.....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.9 ± 0.2 at 25°C
Substrate Solution:
Composition per 10.0mL:
Fructose.......................................................................................10.0g
Preparation of Substrate Solution:  Add fructose to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 10% N 2 . Filter sterilize.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.0g
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.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.7g
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. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically. Before use, neutralize sodium sulfide with sterile hydro-
chloric acid.
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:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O ..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
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 remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under 100%
N 2 gas atmosphere. Add components, except NaHCO 3  solution, sub-
strate solution, Na 2 S·9H 2 O solution, and vitamin solution, to distilled/
deionized water and bring volume to 960.0mL. Mix thoroughly. Adjust
pH to 6.8–7.0. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically add 10.0mL NaHCO 3
solution, 10.0mL substrate solution, 10.0mL Na 2 S·9H 2 O solution, and
10.0mL vitamin solution. Mix thoroughly. Aseptically and anaerobical-
ly distribute into sterile tubes or bottles.
Use:  For the cultivation of Acetobacterium fimetarium DSM 8237 and
Acetobacterium fimetarium DSM 8239.