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Thermodesulfobium Medium (DSMZ Medium 1005)

[所属分类:培养基配方] [发布时间:2021-9-7] [发布人:网站管理员2] [阅读次数:] [返回]

Thermodesulfobium Medium

(DSMZ Medium 1005)

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

Composition per liter:

Na2SO4 ......................................................................................... 2.8g

KH2PO4....................................................................................... 0.78g

K2HPO4....................................................................................... 0.75g

NH4Cl .......................................................................................... 0.5g

MgSO4·7H2O .............................................................................. 0.25g

NaCl ............................................................................................. 0.2g

Na3-EDTA.................................................................................. 0.04g

CaCl2·2H2O................................................................................. 0.03g

Na-acetate .................................................................................. 0.15g

FeSO4·7H2O................................................................................ 0.01g

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

Trace elements solution SL-9 ..................................................10.0mL

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

pH 5.7 ± 0.2 at 25°C

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine-HCl·2H2O ................................................................. 0.25g

Preparation of Cysteine Solution: Add L-cysteine to to distilled/

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

with 100% N2. Filter sterilize.

Trace Elements Solution SL-9:

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 SL-9: 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: Add components, except vitamin and

cysteine solutions, to distilled/deionized water and bring volume to

980.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 3

min. Cool to room temperature while sparging with a gas mixture of

80% N2 + 20% CO2. Adjust the pH to 5.5 with 10N H2SO4. Dispense

under the same gas atmosphere in culture vessels (e.g., 20.0mL of the

medium into 50mL serum bottles). Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to room temperature. Aspetically add vitamin solu

tion. Mix thoroughly. Prior to inoculation change atmosphere to 80%

H2 and 20% CO2 gas mixture. Add the cysteine solution. Adjust pH to

5.5–6.0 if necessary. After inoculation pressurize vials to 0.5 bar over

pressure with 80% H2 and 20% CO2 gas mixture.

Use: For the cultivation of Thermodesulfobium narugense.

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Thermodesulfobium Medium (DSMZ Medium 1005)