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Thermoacetogenium phaeum Medium (DSMZ Medium 880)

[所属分类:培养基配方] [发布时间:2021-9-6] [发布人:网站管理员2] [阅读次数:] [返回]

Thermoacetogenium phaeum Medium

(DSMZ Medium 880)

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

Composition per liter:

KHCO3.......................................................................................... 3.5g

NH4Cl ........................................................................................... 1.0g

NaCl.............................................................................................. 0.6g

KH2PO4......................................................................................... 0.3g

MgCl2·6H2O ................................................................................. 0.1g

CaCl2·2H2O ................................................................................ 0.08g

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

Sodium pyruvate solution........................................................50.0mL

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

Na2S·9H2O solution.................................................................10.0mL

Cysteine solution .....................................................................10.0mL

Trace elements solution .............................................................1.0mL

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

pH 7.0–7.1 at 25°C

Sodium Pyruvate Solution:

Composition per 50.0mL:

Sodium pyruvate........................................................................... 5.0g

Preparation of Sodium Pyruvate Solution: Add sodium pyru

vate to distilled/deionized water and bring volume to 50.0mL. Mix

thoroughly. Sparge with 100% N2. Filter sterilize.

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. Filter sterilize.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.3g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O 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.

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O ..................................................................... 0.3g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O 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.

Trace Elements Solution SL-10:

Composition per liter:

FeCl2·4H2O................................................................................... 1.5g

Na2-EDTA..................................................................................... 0.5g

CoCl2·6H2O ........................................................................... 190.0mg

MnCl2·4H2O .......................................................................... 100.0mg

ZnCl2........................................................................................70.0mg

Na2MoO4·2H2O .......................................................................36.0mg

NiCl2·6H2O ..............................................................................24.0mg

H3BO3 ........................................................................................6.0mg

CuCl2·2H2O ...............................................................................2.0mg

HCl (25% solution)..................................................................10.0mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O

to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized

water and bring volume to 1.0L. Add remaining components. Mix thor

oughly. Adjust pH to 6.5. Sparge with 100% N2. Autoclave for 15 min

at 15 psi pressure–121°C.

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.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Add components, except KHCO3,sodi

um pyruvate solution, cysteine solution, and Na2S·9H2O solution, to

distilled/deionized water and bring volume to 930.0mL. Mix thoroughly.

Gently heat and bring to boiling. Boil for 10 min. Cool to room temper

ature while sparging with 80% N2 + 20% CO2. Add 3.5g KHCO3. Mix

thoroughly while sparging with 80% N2 + 20% CO2 gas atmosphere.

Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaero

bically add 50.0mL sodium pyruvate solution, 10.0mL cysteine solu

tion, and 10.0mL Na2S·9H2O solution. Mix thoroughly. Final pH is 7.0–

7.1. Aseptically and anaerobically distribute into sterile tubes or bottles.

Use: For the cultivation of Thermacetogenium phaeum.

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Thermoacetogenium phaeum Medium (DSMZ Medium 880)