Acetobacterium tundrae Medium (DSMZ Medium 900)

[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 tundrae Medium
(DSMZ Medium 900)
Yeast extract..................................................................................1.0g
MgCl 2 ·6H 2 O................................................................................0.52g
KCl..............................................................................................0.33g
NH 4 Cl .........................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
CaCl 2 ·2H 2 O...................................................................................0.3g
Resazurin ...................................................................................0.5mg
Fructose solution......................................................................30.0mL
NaHCO 3  solution .....................................................................20.0mL
L -Cysteine solution..................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace mineral solution SL-10 ....................................................1.0mL
Seven vitamin solution...............................................................1.0mL
pH 7.0 ± 0.2 at 25°C
L -Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................0.3g
Preparation of  L -Cysteine Solution:  Add  L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
NaHCO 3 Solution:
Composition per 20.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 20.0mL. 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....................................................................................0.3g
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.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Fructose Solution:
Composition per 30.0mL:
Glucose.........................................................................................5.0g
Preparation of Fructose Solution: Add glucose to distilled/de-
ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................200.0mg
Nicotinic acid.........................................................................200.0mg
Vitamin B 12 ............................................................................100.0mg
Calcium pantothenate ............................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
D (+)-Biotin...............................................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. 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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except seven vitamin
solution, NaHCO 3 solution, fructose solution,  L -cysteine-solution, and
Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to
929.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Distribute
into anaerobe tubes or bottles. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add, per liter of medium, 1.0mL
seven vitamin solution, 20.0mL NaHCO 3 30.0mL fructose solution,
10.0mL  L -cysteine-HCl·H 2 O solution, and 10.0mL Na 2 S·9H 2 O solu-
tion. Mix thoroughly. The final pH should be 7.0.
Use: For the cultivation of Acetobacterium tundrae.