Anaerobic Cellulolytic Medium

[所属分类:培养基配方] [发布时间:2019-10-14] [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

Anaerobic Cellulolytic Medium
Composition per liter:
NH 4 Cl ...........................................................................................1.0g
Cellobiose.....................................................................................1.0g
Yeast extract..................................................................................1.0g
MgSO 4 ..........................................................................................0.5g
KCl................................................................................................0.5g
L- Cysteine·HCl·H 2 O .....................................................................0.5g
K 2 HPO 4 .........................................................................................0.4g
Resazurin...................................................................................1.0mg
Wolfe’s mineral solution..........................................................20.0mL
Na 2 CO 3 solution ......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.9 ± 0.1 at 25°C
Wolfe’s Mineral Solution:
Composition per liter
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O ..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.01g
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to approximately 500.0mL of distilled/deionized water and adjust
pH to 6.5 with KOH to dissolve. Bring volume to 1.0L with distilled/de-
ionized water. Add other compounds. Mix thoroughly.
Na 2 CO 3 Solution:
Composition per 100.0mL:
Na 2 CO 3 .......................................................................................10.0g
Preparation of Na 2 CO 3 Solution: Add Na 2 CO 3 to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
Na 2 S·9H 2 O Solution:
Composition per 100.0mL:
Na 2 S·9H 2 O..................................................................................15.0g
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 100.0mL. Mix thoroughly.
Filter sterilize.
Preparation of Medium: Add components, except Na 2 CO 3 solution
and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to
980.0mL. Boil medium under 80% N 2 + 10% CO 2 + 10% H 2 until medium
is colorless. Cool and distribute anaerobically into test tubes in 10.0mL
volumes using 80% N 2 + 10% CO 2 + 10% H 2 . Stopper the tubes anaero-
bically. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room tem-
perature. Aseptically add 10.0mL of sterile Na 2 CO 3 solution and 10.0mL
of sterile Na 2 S·9H 2 O solution to each tube. Mix thoroughly.
Use: For the cultivation and maintenance of microorganisms that can
utilize cellobiose as sole carbon source, such as Clostridium cellulo-
vorans.