Acetobacterium dehalogenans Medium (DSMZ Medium 787)

[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 dehalogenans Medium
(DSMZ Medium 787)
Composition per liter:
NaHCO 3 ......................................................................................10.0g
Yeast extract..................................................................................2.0g
NH 4 Cl ...........................................................................................1.0g
K 2 HPO 4 .......................................................................................0.45g
KH 2 PO 4 .......................................................................................0.33g
MgSO 4 ·7H 2 O................................................................................0.1g
Resazurin ...................................................................................1.0mg
NaHCO 3  solution .....................................................................30.0mL
Na 2 CO 3  solution.......................................................................20.0mL
Trace elements solution ...........................................................20.0mL
Vitamin solution.......................................................................20.0mL
Na-syringate soltuion...............................................................10.0mL
Cysteine solution......................................................................10.0mL
pH 7.4 ± 0.2 at 25°C
NaHCO 3 Solution:
Composition per 100.0mL:
NaHCO 3 ......................................................................................10.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Filter sterilize.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................0.3g
Preparation of 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. Cool to 25°C.
Na-syringate Solution:
Composition per 20.0mL:
Na-syringate..................................................................................1.2g
Preparation of Na-syringate Solution: Add Na-syringate to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Sparge with 100% N 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.
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% N 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solu-
tion, Na 2 CO 3  solution, vitamin solution, Na-syringate solution, and
cysteine solution, to distilled/deionized water and bring volume to
900.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 5
min. Cool while sparging with 80% N 2 + 20% CO 2 . Distribute 9.0mL al-
iquots into serum bottles. Autoclave under 80% N 2 + 20% CO 2  for 15
min at 15 psi pressure–121°C. Aseptically and anaerobically add approx-
imately 0.20mL sterile Na 2 CO 3  solution to each 9.0mL of medium so
that pH is adjusted to 7.4. For every 9.0mL of medium inject 1.0mL
NaHCO 3  solution, 0.15mL Na-syringate solution, 0.2mL vitamin solu-
tion, and 0.17mL cysteine solution.
Use:  For the cultivation of Acetobacterium dehalogenans.