Desulfitobacterium PCE 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 PCE Medium

培养基配方
Composition per 1001.0mL:
(NH4)H2PO4 ..................................................................2.88g
MgSO4 ·7H2O..................................................................0.1g
Yeast extract.................................................................0.1g
Ca(NO3)2 ·4H2O .............................................................0.05g
Resazurin....................................................................0.1mg
NaHCO3 solution.............................................................50.0mL
KOH solution................................................................20.0mL
Sodium fumarate solution....................................................20.0mL
Sodium- L -lactate solution.................................................20.0mL
Wolfe’s vitamin solution...................................................10.0mL
Seven vitamin solution ......................................................1.0mL
Selenite-tungstate solution..................................................1.0mL
Wolfe’s mineral solution....................................................1.0mL
pH 7.0–7.2 at 25°C
NaHCO3 Solution:
Composition per 50.0mL:
NaHCO3 .......................................................................2.5g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
KOH Solution:
Composition per 20.0mL:
KOH.............................................................................2.0g
Preparation of KOH Solution:  Add KOH to distilled/deionized
water and bring volume to 20.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Sodium Fumarate Solution:
Composition per 20.0mL:
Sodium fumarate..................................................................3.2g
Preparation of Sodium Fumarate Solution:  Add sodium fu-
marate to distilled/deionized water and bring volume to 20.0mL. Mix
thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Sodium  L -Lactate Solution:
Composition per 20.0mL:
Sodium  L -lactate ..............................................................3.2g
Preparation of Sodium  L -Lactate Solution:  Add sodium  L -lac-
tate to distilled/deionized water and bring volume to 20.0mL. Mix
thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pres-
sure–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.
Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine·HCl..................................................................0.3g
Thiamine·HCl ...................................................................0.2g
Nicotinic acid...................................................................0.2g
Calcium  DL -pantothenate........................................................0.1g
Vitamin B 12 ....................................................................0.1g
p-Aminobenzoic acid...........................................................80.0mg
Biotin .......................................................................20.0mg
Preparation of Seven Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Selenite-Tungstate Solution:
Composition per liter:
NaOH...........................................................................0.5g
Na2WO4 ·2H2O.................................................................4.0mg
Na2SeO3 ·5H2O................................................................3.0mg
Preparation of Selenite-Tungstate Solution:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
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 and dispense medium under 80% N2
+ 20% CO2 gas mixture. Add components, except NaHCO3 solution, KOH
solution, sodium fumarate solution, sodium- L -lactate solution, Wolfe’s vita-
min solution, and seven vitamins solution, to distilled/deionized water and
bring volume to 880.0mL. Mix thoroughly. Adjust pH to 7.0–7.2. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 50.0mL of sterile NaHCO3 solution, 20.0mL of sterile
KOH solution, 20.0mL of sterile sodium fumarate solution, 20.0mL of ster-
ile sodium- L -lactate solution, 10.0mL of sterile Wolfe’s vitamin solution,
and 1.0mL of sterile seven vitamins solution. Mix thoroughly. Aseptically
and anaerobically distribute into sterile tubes or bottles.
Use: For the cultivation of Desulfitobacterium species.