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Sulfurimonas paralvinella Medium (DSMZ Medium 1053)

[所属分类:培养基配方] [发布时间:2021-8-31] [发布人:网站管理员2] [阅读次数:] [返回]

Sulfurimonas paralvinella Medium

(DSMZ Medium 1053)

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

Composition per liter:

NaCl ........................................................................................... 20.0g

Sulfur, elemental......................................................................... 10.0g

MgSO4·7H2O................................................................................ 4.0g

MgCl2·6H2O ................................................................................. 3.0g

Na2S2O3·5H2O ............................................................................. 1.0g

NaNO3 .......................................................................................... 1.0g

CaCl2·2H2O .................................................................................. 0.8g

KCl ............................................................................................. 0.33g

NH4Cl ........................................................................................ 0.25g

K2HPO4....................................................................................... 0.09g

KH2PO4....................................................................................... 0.07g

Fe2(SO4)3·H2O ............................................................................ 0.01g

Resazurin .................................................................................. 0.5mg

Trace elements solution ...........................................................10.0mL

Bicarbonate solution ................................................................10.0mL

Vitamin solution.......................................................................10.0mL

Selenite-tungstate solution.........................................................1.0mL

pH 6.8 ± 0.2 at 25°C

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.

Vitamin Solution:

Composition per liter:

Pyridoxine-HCl........................................................................10.0mg

Thiamine-HCl·2H2O..................................................................5.0mg

Riboflavin ..................................................................................5.0mg

Nicotinic acid.............................................................................5.0mg

D-Ca-pantothenate......................................................................5.0mg

p-Aminobenzoic acid.................................................................5.0mg

Lipoic acid .................................................................................5.0mg

Biotin .........................................................................................2.0mg

Folic acid....................................................................................2.0mg

Vitamin B12 ................................................................................0.1mg

Preparation of Vitamin Solution: Add components to distilled/

deionized water and bring volume to 1.0L. Mix thoroughly. Sparge

with 80% H2 + 20% CO2. Filter sterilize.

Bicarbonate Solution:

Composition per 10.0mL:

NaHCO3 ....................................................................................... 1.0g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge

with 20% CO2 + 80% N2. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to room temperature.

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: Steam sulfur for 3 hr on each of 3 succes

sive days. Add the sulfur to the culture vessels.Add components, except

vitamin solution, sulfur, and bicarbonate solution, to distilled/deionized

water and bring volume to 975.0mL. Mix thoroughly. Gently heat and

bring to boiling. Boil for 3 min. Cool to room temperature under 80%

H2 + 20% CO2. Adjust pH to 6.8 with NaOH. Dispense under same gas

atmosphere into the culture vessels containing the sulfur (up to volume

of 20%). Autoclave for 20 min at 6 psi pressure–110°C. Aseptically add

vitamin and bicarbonate solutions. Adjust the pH to 6.5. After inocula

tion pressurize vessels to 2 bar overpressure with 80% H2 + 20% CO2

gas mixture.

Use: For the cultivation of Sulfurimonas paralvinella.

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Sulfurimonas paralvinella Medium (DSMZ Medium 1053)