Acetobacterium sp. KoMAc1 Medium (DSMZ Medium 478)

[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. KoMAc1 Medium
(DSMZ Medium 478)
Composition per 1001.0mL:
Solution A..............................................................................870.0mL
Solution C..............................................................................100.0mL
Solution D................................................................................10.0mL
Solution E (Vitamin solution)..................................................10.0mL
Solution F.................................................................................10.0mL
Solution B (Trace element solution SL-10)...............................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
Na 2 SO 4 ..........................................................................................3.0g
NaCl..............................................................................................1.0g
Yeast extract..................................................................................0.5g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O..................................................................................0.4g
NH 4 Cl ...........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
Resazurin ...................................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly.
Solution B (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 Solution B (Trace Elements Solution SL-10):
Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min
at 15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of Solution C: Add NaHCO 3 to distilled/deionized
water and bring volume to 100.0mL Mix thoroughly. Filter sterilize.
Flush with 80% N 2  + 20% CO 2 to remove dissolved oxygen.
Solution D:
Composition per 10.0mL:
Methoxyacetate.............................................................................0.9g
Preparation of Solution D: Add methoxyacetate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E (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 ..............................................................................0.10mg
Solution E (Vitamin Solution): Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution F:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.4g
Preparation of Solution F:  Add Na 2 S·9H 2 O to distilled/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.
Preparation of Medium: Gently heat solution A and bring to boil-
ing. Boil solution A for a few minutes. Cool to room temperature. Gas
with 80% N 2  + 20% CO 2 gas mixture to reach a pH below 6. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se-
quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solu-
tion D, 10.0mL solution E, and 10.0mL solution F. Distribute
anaerobically under 80% N 2  + 20% CO 2 into appropriate vessels. Ad-
dition of 10–20mg sodium dithionite  per liter from a 5% (w/v) solu-
tion, freshly prepared under N 2  and filter-sterilized, may stimulate
growth.
Use: For the cultivation of Acetobacterium sp.