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Syntrophothermus Medium (DSMZ Medium 870)

[所属分类:培养基配方] [发布时间:2021-9-1] [发布人:网站管理员2] [阅读次数:] [返回]

Syntrophothermus Medium

(DSMZ Medium 870)

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

Composition per liter:

NaHCO3 ........................................................................................ 2.5g

NH4Cl ......................................................................................... 0.54g

MgCl2·6H2O.................................................................................. 0.2g

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

KH2PO4....................................................................................... 0.14g

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

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

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

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

Substrate solution.....................................................................10.0mL

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

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

pH 7.0 ± 0.2 at 25°C

Substrate Solution:

Composition per 10.0mL:

Na-crotonate................................................................................ 0.86g

Preparation of Substrate Solution: AddNa-crotonate to distilled/

deionized water and bring volume to 10.0mL. 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.

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

to room temperature.

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.

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.

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.

Preparation of Medium: Prepare and dispense medium under 80% N2

+ 20% CO2 gas atmosphere. Add components, except cysteine solution,

Na2S·9H2O solution, and substrate solution, to distilled/deionized water and

bring volume to 970.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with

80% N2 + 20% CO2 for 30 min. Distribute into Hungate tubes or serum bot

tles. Autoclave for 15 min at 15 psi pressure–121°C. For each 10.0mL me

dium, aseptically and anaerobically add 0.1mL cysteine solution, 0.1mL

Na2S·9H2O solution, and 0.1mL substrate solution. Mix thoroughly.

Use: For the cultivation of Syntrophothermus lipocalidus DSM 12680.

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Syntrophothermus Medium (DSMZ Medium 870)