Clostridium cellulovorans Medium

[所属分类:培养基配方] [发布时间:2020-5-9] [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

Clostridium cellulovorans Medium

培养基配方
Composition per liter:
K2HPO4 ·3H2O..............................................................1.0g
NH4Cl .....................................................................1.0g
KCl........................................................................0.5g
MgSO4 ·7H2O...............................................................0.5g
Pancreatic digest of casein................................................0.5g
Yeast extract..............................................................0.5g
L -Cysteine·HCl·H2O ....................................................0.15g
Resazurin ................................................................1.0mg
Cellulose, MN 300 or cellobiose solution ................................50.0mL
Na2CO3  solution.........................................................30.0mL
Rumen fluid, clarified...................................................20.0mL
Na2S·9H2O solution......................................................20.0mL
Trace elements solution SL-10 ............................................1.0mL
pH 7.0 ± 0.2 at 25°C
Cellobiose Solution:
Composition per 50.0mL:
Cellulose, MN 300 or D- cellobiose ........................................5.0g
Preparation of Cellobiose Solution: Add cellulose or cellobiose
to distilled/deionized water and bring volume to 50.0mL. Mix thor-
oughly. Sparge under 100% N2 gas for 3 min. Filter sterilize. Store un-
der N2 gas.
Na2CO3 Solution:
Composition per 30.0mL:
Na2CO3 ....................................................................1.0g
Preparation of Na2CO3 Solution:  Add Na2CO3 to distilled/deion-
ized water and bring volume to 30.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 20.0mL:
Na2S·9H2O.................................................................0.15g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O...............................................................1.5g
CoCl2 ·6H2O............................................................190.0mg
MnCl2 ·4H2O ...........................................................100.0mg
ZnCl2 ...................................................................70.0mg
Na2MoO4 ·2H2O...........................................................36.0mg
NiCl2 ·6H2O.............................................................24.0mg
H3BO3 ....................................................................6.0mg
CuCl2 ·2H2O..............................................................2.0mg
HCl(25% solution)........................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
Preparation of Medium: Add components, except cellobiose solu-
tion, Na2CO3  solution, and Na2S·9H2O solution, to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 50.0mL of sterile cellobiose solution, 30.0mL of
sterile Na2CO3  solution, and 20.0mL of sterile Na2S·9H2O solution.
Mix thoroughly. Aseptically and anaerobically distribute into sterile
screw-capped bottles.
Use:  For the cultivation and maintenance of Clostridium cellulo-
vorans.