Desulfitobacterium PCE II Medium (DSMZ Medium 1062)

[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 II Medium
(DSMZ Medium 1062)

培养基配方
Composition per liter:
NaCl.................................................................................1.0g
KCl..................................................................................0.5g
MgCl2 ·6H2O ........................................................................0.4g
NH4Cl ..............................................................................0.25g
KH2PO4 ..............................................................................0.2g
CaCl2 ·2H2O .......................................................................0.15g
Resazurin ..........................................................................0.5mg
Trace elements solution............................................................10.0mL
Pyruvate solution..................................................................10.0mL
Fumarate solution .................................................................10.0mL
Yeast extract solution.............................................................10.0mL
Ferrous sulfate solution...........................................................10.0mL
NaHCO3  solution...................................................................10.0mL
Selenite/tungstate solution.........................................................1.0mL
Vitamin solution....................................................................1.0mL
pH 7.5 ± 0.2 at 25°C
Ferrous Sulfate Solution:
Composition per 10.0mL:
FeSO4 ·7H2O.......................................................................22.0mg
Preparation of Ferrous Sulfate Solution:  Add FeSO4 ·7H2O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Pyruvate Solution:
Composition per 10.0mL:
Na-pyruvate ........................................................................4.5g
Preparation of Pyruvate Solution:  Add Na-pyruvate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Fumarate Solution:
Composition per 10.0mL:
Na2 -fumarate ......................................................................6.5g
Preparation of Fumarate Solution:  Add Na2 -fumarate 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.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract ......................................................................2.0g
Preparation of Yeast Extract Solution:  Add yeast extract 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.
NaHCO3 Solution :
Composition per 10.0mL:
NaHCO3 .............................................................................3.0g
Preparation of NaHCO3 Solution:  Add NaHCO3  to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
Vitamin Solution:
Nicotinamide........................................................................9.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Riboflavin .........................................................................5.0mg
Thiamine·HCl ......................................................................5.0mg
Biotin .............................................................................2.0mg
Folic acid..........................................................................2.0mg
Cyanocobalamin .....................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Trace Elements Solution:
Composition per liter:
MgSO4 ·7H2O........................................................................3.0g
Nitrilotriacetic acid...............................................................1.5g
NaCl................................................................................1.0g
MnSO4 ·2H2O........................................................................0.5g
CoSO4 ·7H2O.......................................................................0.18g
ZnSO4 ·7H2O.......................................................................0.18g
CaCl2 ·2H2O........................................................................0.1g
FeSO4 ·7H2O........................................................................0.1g
NiCl2 ·6H2O......................................................................0.025g
KAl(SO4)2 ·12H2O..................................................................0.02g
H3BO3 .............................................................................0.01g
Na2MoO4 ·4H2O ....................................................................0.01g
CuSO4 ·5H2O.......................................................................0.01g
Na2SeO3 ·5H2O.....................................................................0.3mg
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water 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.
Preparation of Medium:  Add components, except bicarbonate,
iron sulfate, vitamins, and organic substrates, to distilled/deionized wa-
ter and bring volume to 940.0mL. Gently heat and bring to boiling.
Boil for 1 min. Mix thoroughly. Cool to room temperature while sparg-
ing with 80% N2  + 20% CO2 . Dispense into culture vessels under an
atmosphere of 80% N2  + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Add ferrous iron sulfate, vitamins, pyruvate, fumarate,
and yeast extract from sterile anoxic solutions. Adjust the final pH of
the medium to 7.5. Addition of 10–20 mg sodium dithionite per liter
(e.g., from 5% (w/v) solution, freshly prepared under N2  and filter ster-
ilized) may stimulate growth at the beginning.
Use: For the cultivation of Desulfitobacterium spp.