Butyrivibrio Species Medium

[所属分类:培养基配方] [发布时间:2020-3-24] [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

Butyrivibrio Species Medium

培养基配方
Composition per 1001.0mL:
Na2CO3 ............................................................4.0g
Pancreatic digest of casein........................................2.0g
Yeast extract......................................................2.0g
K2HPO4 ............................................................0.3g
Hemin ............................................................1.0mg
Resazurin ........................................................1.0mg
Rumen fluid, clarified..........................................150.0mL
Minerals solution................................................75.0mL
Carbohydrate solution............................................20.0mL
L -Cysteine·HCl·H2O solution...................................10.0mL
Na2S·9H2O solution..............................................10.0mL
Volatile fatty acid mixture ......................................3.1mL
pH 6.7 ± 0.2 at 25°C
Minerals Solution:
Composition per liter:
NaCl..............................................................12.0g
KH2PO4 ............................................................6.0g
(NH4)2SO4 .........................................................6.0g
MgSO4 ·7H2O.......................................................2.5g
CaCl2 ·2H2O.......................................................1.6g
Preparation of Minerals Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O .............................................0.25g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -cys-
teine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% CO2 . Autoclave for 15 min at 15 psi
pressure–121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.......................................................0.25g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Carbohydrate Solution:
Composition per 20.0mL:
Glucose .........................................................1.0g
Cellobiose ......................................................1.0g
Glycerol ........................................................1.0g
Maltose..........................................................1.0g
Starch, soluble..................................................1.0g
Preparation of Carbohydrate Solution : Add components to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Sparge under 100% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Volatile Fatty Acid Mixture:
Composition per 7.75mL:
Acetic acid ....................................................4.25mL
Propionic acid..................................................1.50mL
Butyric acid.....................................................1.0mL
DL -2-Methyl butyric acid.......................................0.25mL
iso-Butyric acid................................................0.25mL
iso-Valeric acid ...............................................0.25mL
n-Valeric acid..................................................0.25mL
Preparation of Volatile Fatty Acid Mixture: Combine compo-
nents. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under
100% CO2 . Add components, except carbohydrate solution, Na2CO3 ,
L -cysteine·HCl·H2O solution, and Na2S·9H2O solution, to distilled/de-
ionized water and bring volume to 960.0mL Mix thoroughly. Gently
heat and bring to boiling. Continue boiling for 5 min. Cool to room
temperature while sparging with 100% CO2 . Add Na2CO3 . Continue
sparging with 100% CO2 until pH reaches 6.8. Distribute into rubber-
stoppered tubes under 100% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Aseptically and anaerobically add 20.0mL of sterile car-
bohydrate solution, 10.0mL of sterile  L -cysteine·HCl·H2O solution,
and 10.0mL of sterile Na2S·9H2O solution or, using a syringe, inject
the appropriate amount of sterile carbohydrate solution, sterile
Na2S·9H2O solution, and sterile  L -cysteine·HCl·H2O solution into in-
dividual tubes containing medium.
Use: For the cultivation of Butyrivibrio species.