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Thauera aromatica AR-1 Medium (DSMZ Medium 855)

[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]

Thauera aromatica AR-1 Medium

(DSMZ Medium 855)

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

Composition per 992.0mL:

Solution A ..............................................................................870.0mL

Solution C ..............................................................................100.0mL

Solution D ................................................................................10.0mL

Solution E (Vitamin solution) ..................................................10.0mL

Solution B (Trace elements solution SL-10) .............................1.0mL

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

pH 7.2 ± 0.2 at 25°C

Solution A:

Composition per 870.0mL:

Na2SO4.......................................................................................... 3.0g

NaCl .............................................................................................. 1.0g

KNO3 ............................................................................................ 0.6g

KCl................................................................................................ 0.5g

MgCl2·6H2O.................................................................................. 0.4g

NH4Cl ........................................................................................... 0.3g

KH2PO4......................................................................................... 0.2g

CaCl2·2H2O................................................................................. 0.15g

Resazurin ...................................................................................1.0mg

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 870.0mL. Mix thoroughly.

Solution B (Trace Elements Solution SL-10):

Composition per liter:

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

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 Solution B (Trace Elements Solution SL-10):

Add FeCl2·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis

tilled/deionized water and bring volume to 1.0L. Add remaining com

ponents. Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min

at 15 psi pressure–121°C.

Solution C:

Composition per 100.0mL:

NaHCO3 ........................................................................................ 5.0g

Preparation of Solution C: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Flush with 80% N2 + 20% CO2 to remove dissolved oxygen.

Solution D:

Composition per 10.0mL:

Na-benzoate .................................................................................. 0.7g

Preparation of Solution D: Add Na-benzoate to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with

100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Solution E (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 .............................................................................. 0.10mg

Solution E (Vitamin Solution): Add components to distilled/de

ionized water and bring volume to 1.0L. Mix thoroughly. Sparge with

100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

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.

Preparation of Medium: Gently heat solution A and bring to boil

ing. Boil solution A for a few minutes. Cool to room temperature. Gas

with 80% N2 + 20% CO2 gas mixture to reach a pH below 6. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se

quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solu

tion D, 10.0mL solution E, and 1.0mL sterile selenite-tungstate

solution. Distribute anaerobically under 80% N2 + 20% CO2 into ap

propriate vessels. Addition of 10–20mg sodium dithionite per liter

from a 5% (w/v) solution, freshly prepared under N2 and filter-steril

ized, may stimulate growth.

Use: For the anaerobic cultivation of Thauera aromatica.

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Thauera aromatica AR-1 Medium (DSMZ Medium 855)