Fibrobacter Medium

[所属分类:培养基配方] [发布时间:2020-7-16] [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

Fibrobacter Medium

培养基配方
Composition per 1020.0mL:
Cellobiose ...............................................................4.0g
Na2CO3 ...................................................................4.0g
Pancreatic digest of casein...............................................1.0g
NaCl......................................................................0.6g
Yeast extract.............................................................0.5g
K2HPO4 ...................................................................0.3g
KH2PO4 ...................................................................0.3g
(NH4)2SO4 ................................................................0.3g
MgSO4 ·7H2O.............................................................0.12g
CaCl2 ·2H2O.............................................................0.08g
Resazurin ...............................................................1.0mg
Vitamin solution.........................................................20.0mL
Na2S·9H2O solution.....................................................10.0 ML
L -Cysteine·HCl·H2O solution ..........................................10.0mL
VFA solution.............................................................4.65mL
Trace elements solution...................................................1.0mL
pH 6.6 ± 0.2 at 25°C
Vitamin Solution:
Composition per 100.0mL:
Calcium  D -(+)-pantothenate ............................................20.0mg
Lipoic acid..............................................................20.0mg
Nicotinamide.............................................................20.0mg
Pyridoxal·HCl...........................................................20.0mg
Pyridoxamine·2HCl.......................................................20.0mg
Riboflavin...............................................................20.0mg
Thiamine·HCl ...........................................................20.0mg
p-Aminobenzoic acid......................................................1.0mg
Biotin...................................................................1.0mg
Cyanocobalamin...........................................................1.0mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly.
Na 2 S·9H 2 O 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% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O...................................................0.25g
Preparation of  L -Cysteine·HCl·H2O Solution:  Add  L -cyste-
ine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi
pressure–121°C.
Trace Elements Solution:
Composition per liter:
FeSO4 ·7H2O...............................................................2.0g
CoCl2 ·6H2O...............................................................0.2g
H3BO3 .....................................................................0.2g
ZnSO4 ·7H2O...............................................................0.1g
MnCl2 ·4H2O .............................................................0.03g
Na2MoO4 ·2H2O............................................................0.03g
NiCl2 ·6H2O..............................................................0.02g
CuCl2 ·2H2O..............................................................0.01g
Preparation of Trace Elements Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
VFA Solution:
Composition per 310.0mL:
Acetic acid..............................................................17.0mL
Propionic acid............................................................6.0mL
n-Butyric acid ...........................................................4.0mL
Isobutyric acid ..........................................................1.0mL
Isovaleric acid...........................................................1.0mL
DL -α-Methylbutyric acid ................................................1.0mL
n-Valeric acid............................................................1.0mL
Preparation of VFA Solution: Add volatile fatty acids to approx-
imately 200.0mL of distilled/deionized water. Mix thoroughly. Adjust
pH to 7.0 with NaOH pellets. Bring volume to 310.0mL with distilled/
deionized water.
Preparation of Medium: Prepare and dispense medium under 100%
CO2 . Add components, except Na2CO3 , to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling.
Continue boiling for 3 min. Cool to room temperature while sparging with
100% CO 2 . Add Na 2 CO 3 . Mix thoroughly. Continue sparging with 100%
CO 2 for 10 min. Anaerobically distribute into anaerobic tubes. Autoclave
for 15 min at 15 psi pressure–121°C. Prior to inoculation, aseptically and
anaerobically add 10.0mL of sterile Na2S·9H2O solution and 10.0mL of
sterile  L -cysteine·HCl·H2O solution per liter of medium. Mix thoroughly.
Adjust pH to 6.6.
Use: For the cultivation of Fibrobacter intestinalis and Fibrobacter
succinogenes.