CP Medium for Coprothermobacter proteolyticus

[所属分类:培养基配方] [发布时间:2020-5-26] [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

CP Medium for Coprothermobacter proteolyticus

培养基配方
Composition per 1010.0mL:
NaHCO3 ........................................................................................8.4g
Pancreatic digest of casein............................................................2.0g
Yeast extract..................................................................................2.0g
MgCl2 ·6H2O .................................................................................1.0g
NH4Cl ...........................................................................................1.0g
CaCl2 ·2H2O ..................................................................................0.4g
K2HPO4 ·3H2O ..............................................................................0.4g
Resazurin ...................................................................................0.5mg
Gelatin solution......................................................................100.0mL
Na2S·9H2O solution.................................................................10.0mL
Wolfe’s mineral solution..........................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Gelatin Solution:
Composition per 100.0mL:
Gelatin...........................................................................................3.0g
Preparation of Gelatin Solution:  Add gelatin to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O....................................................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO4 ·7H2O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO4 ·2H2O................................................................................0.5g
CoCl2 ·6H2O ..................................................................................0.1g
ZnSO4 ·7H2O.................................................................................0.1g
CaCl2 ·2H2O...................................................................................0.1g
FeSO4 ·7H2O..................................................................................0.1g
NiCl2 ·6H2O...............................................................................0.025g
KAl(SO4)2 ·12H2O.......................................................................0.02g
CuSO4 ·5H2O...............................................................................0.01g
H3BO3 .........................................................................................0.01g
Na2MoO4 ·2H2O ..........................................................................0.01g
Na2SeO3 ·5H2O...........................................................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
Preparation of Medium: Prepare medium anaerobically under
100% CO2 . Add components, except gelatin solution, Na 2 S·9H 2 O so-
lution, and Wolfe’s mineral solution, to distilled/deionized water and
bring volume to 880.0mL. Mix thoroughly. Gently heat and bring to
boiling. Cool to room temperature while sparging with 100% CO2 .
Sparge with 100% CO2 for 20 min. Adjust pH to 7.0. Autoclave for
15 min at 15 psi pressure–121°C. Aseptically and anaerobically add
100.0mL of sterile gelatin solution, 10.0mL of sterile Na2S·9H2O so-
lution, and 10.0mL of sterile Wolfe’s mineral solution to each tube.
Mix thoroughly.
Use:  For the cultivation of Coprothermobacter proteolyticus.