Desulfitobacterium dehalogenans Medium

[所属分类:培养基配方] [发布时间:2020-6-4] [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

Desulfitobacterium dehalogenans Medium

培养基,培养基配方
Composition per liter:
KH2PO4 ....................................................................5.4g
Sodium pyruvate............................................................2.2g
3-Chloro-4-hydroxyphenylacetic acid........................................1.9g
Yeast extract..............................................................1.0g
NH4Cl .....................................................................0.5g
MgCl2 ·6H2O ............................................................90.0mg
CaCl2 ...................................................................25.0mg
Reducing solution........................................................20.0mL
Wolfe’s vitamin solution................................................10.0mL
Modified Wolfe’s mineral solution........................................5.0mL
pH 7.5 ± 0.2 at 25°C
Reducing Solution:
Composition per liter:
L -Cysteine·HCl·H2O....................................................12.5g
Na2S·9H2O...............................................................12.5g
NaOH.....................................................................8.0g
Preparation of Reducing Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 100% N2 . Anaerobically distribute into anaerobic tubes. Auto-
clave for 15 min at 15 psi pressure–121°C.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl.......................................................10.0mg
p-Aminobenzoic acid....................................................5.0mg
Lipoic acid............................................................5.0mg
Nicotinic acid.........................................................5.0mg
Riboflavin.............................................................5.0mg
Thiamine·HCl .........................................................5.0mg
Calcium  DL -pantothenate..............................................5.0mg
Biotin.................................................................2.0mg
Folic acid ............................................................2.0mg
Vitamin B 12 ..........................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Modified Wolfe’s Mineral Solution:
Composition per liter:
MgSO4 ·7H2O............................................................3.0g
Nitrilotriacetic acid...................................................1.5g
NaCl....................................................................1.0g
MnSO4 ·H2O.............................................................0.5g
CaCl2 ..................................................................0.1g
CoCl2 ·6H2O............................................................0.1g
FeSO4 ·7H2O............................................................0.1g
ZnSO4 ·7H2O............................................................0.1g
AlK(SO4)2 ·12H2O......................................................0.01g
CuSO4 ·5H2O...........................................................0.01g
H3BO3 .................................................................0.01g
Na2MoO4 ·2H2O.........................................................0.01g
Na2SeO3 ...............................................................0.01g
NaWO4 ·2H2O ..........................................................0.01g
NiC12 ·6H2O...........................................................0.01g
Preparation of Modified Wolfe’s Mineral Solution: Add
nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH
to 6.5 with KOH. Add remaining components one at a time. Add dis-
tilled/deionized water to 1.0L. Adjust pH to 6.8.
Preparation of Medium: Prepare and dispense medium under
100% N2 . Add components, except reducing solution, to distilled/de-
ionized water and bring volume to 980.0mL. Mix thoroughly. Gently
heat and bring to boiling. Continue boiling for 3 min. Cool to room
temperature while sparging with 100% N2 . Add reducing solution. Mix
thoroughly. Adjust pH to 7.5. Anaerobically distribute into anaerobic
tubes. Autoclave for 15 min at 15 psi pressure–121°C.
Use: For the cultivation of Desulfitobacterium dehalogenans.