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Thermosulfidibacter Medium (DSMZ Medium 1092)

[所属分类:培养基配方] [发布时间:2021-9-8] [发布人:网站管理员2] [阅读次数:] [返回]

Thermosulfidibacter Medium

(DSMZ Medium 1092)

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

Composition per liter:

Sea salts, Sigma .......................................................................... 30.0g

Sulfur, elemental ........................................................................... 3.0g

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

Vitamin solution.......................................................................20.0mL

Sulfide solution ........................................................................10.0mL

Yeast extract solution...............................................................10.0mL

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

Wolfe's mineral elixir.................................................................1.0mL

pH 7.1 ± 0.2 at 25°C

Sulfide Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

Preparation of Sulfide Solution: Add Na2S·9H2O to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave

under 100% N2 for 15 min at 15 psi pressure–121°C. Cool to room

temperature.

Wolfe’s Mineral Elixir:

Composition per liter:

MgSO4·7H2O .............................................................................. 30.0g

NaCl ............................................................................................ 10.0g

MnSO4·2H2O ................................................................................ 5.0g

(NH4)2NiSO4·6H2O ...................................................................... 2.8g

CoCl2·6H2O .................................................................................. 1.8g

ZnSO4·7H2O ................................................................................. 1.8g

FeSO4·7H2O.................................................................................. 1.0g

CaCl2·2H2O................................................................................... 1.0g

KAl(SO4)2·12H2O....................................................................... 0.18g

CuSO4·5H2O ................................................................................. 0.1g

H3BO3 ........................................................................................... 0.1g

Na2MoO4·2H2O ............................................................................ 0.1g

Na2SeO4 ........................................................................................ 0.1g

Na2WO4·2H2O .............................................................................. 0.1g

Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of dis

tilled/deionized water to 1.0 with dilute H2SO4. Add remaining com

ponents one at a time. Mix throughly to dissolve.

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 ........................................................................................ 2.5g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

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

with gas mixture of 80% N2 + 20% CO2. Filter sterilize.

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 gas. Autoclave for 15 min at 15 psi pressure–

121°C.

Preparation of Medium: Add components, except vitamin, bicar

bonate, sulfide, and yeast extract solutions, to distilled/deionized water

and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring

to boiling. Boil for 1 min. Cool to room temperature while sparging

with a gas mixture of 80% H2 + 20% CO2. Adjust the pH to 5.5 with

10N H2SO4. Dispense under the same gas atmosphere in culture ves

sels (balch tubes). Autoclave for 20 min at 6 psi pressure–110°C. Cool

to room temperature. Aspetically add vitamin, bicarbonate, sulfide,

and yeast extract solutions. Mix thoroughly. Prior to inoculation

change atmosphere to 80% H2 and 20% CO2 gas mixture. Adjust pH to

7.0–7.2. After inoculation pressurize vials to 2.0 bar overpressure with

80% H2 and 20% CO2 gas mixture.

Use: For the cultivation of Thermosulfidibacter spp.

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Thermosulfidibacter Medium (DSMZ Medium 1092)