Clostridium neopropionicum Medium

[所属分类:培养基配方] [发布时间:2020-5-13] [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 neopropionicum Medium

培养基配方
Composition per liter
KHCO3 ......................................................................4.0g
Ethanol.....................................................................1.0g
NH4Cl ......................................................................1.0g
NaCl........................................................................0.6g
Pancreatic digest of casein.................................................0.5g
Yeast extract...............................................................0.5g
KH2PO4 .....................................................................0.3g
MgCl2 ·6H2O................................................................0.1g
CaCl2 ·2H2O...............................................................0.08g
Resazurin .................................................................1.0mg
Trace elements solution ..................................................10.0mL
Vitamin solution..........................................................10.0mL
L -Cysteine·HCl·H2O solution ............................................10.0mL
Na2S·9H2O solution........................................................10.0mL
pH 6.8 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.......................................................12.8g
FeCl3 ·6H2O................................................................1.35g
NaCl.........................................................................1.0g
NiCl2 ·6H2O................................................................0.12g
MnCl2 ·4H2O.................................................................0.1g
CaCl2 ·2H2O.................................................................0.1g
ZnCl2 .......................................................................0.1g
Na2SeO3 ·5H2O.............................................................0.026g
CuCl2 ·2H2O ..............................................................0.025g
CoCl2 ·6H2O ..............................................................0.024g
Na2MoO4 ·2H2O ............................................................0.024g
H3BO3 ......................................................................0.01g
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to approximately 500.0mL of distilled/deionized water. Dissolve
by adding KOH and adjust pH to 6.5. Add remaining components.
Bring volume to 1.0L with additional distilled/deionized water. Adjust
pH to 7.0 with KOH.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl............................................................10.0mg
Calcium  DL -pantothenate...................................................5.0mg
Lipoic acid.................................................................5.0mg
Nicotinic acid..............................................................5.0mg
p-Aminobenzoic acid.........................................................5.0mg
Riboflavin..................................................................5.0mg
Thiamine·HCl...............................................................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. Filter ster-
ilize. Sparge with 80% N2 + 20% CO2 .
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl ..........................................................0.25g
Preparation of  L -Cysteine·HCl Solution: Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 80% N2 + 20% CO2 for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O...................................................................0.3g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Preparation of Medium: Add components, except vitamin solu-
tion,  L -cysteine·HCl·H2O solution, and Na2S·9H2O solution, to dis-
tilled/deionized water and bring volume to 960.0mL. Mix thoroughly.
Sparge under 80% N2 + 20% CO2 for 3–4 min. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL of
sterile vitamin solution, 10.0mL of sterile  L -cysteine·HCl·H2O solu-
tion, and 10.0mL of sterile Na2S·9H2O solution. Mix thoroughly.
Aseptically and anaerobically distribute into sterile screw-capped bot-
tles under 80% N2 + 20% CO2 .
Use:  For the cultivation and maintenance of Clostridium neopropioni-
cum