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Smithella Medium (DSMZ Medium 1030)

[所属分类:培养基配方] [发布时间:2021-8-18] [发布人:网站管理员2] [阅读次数:] [返回]

Smithella Medium

(DSMZ Medium 1030)

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

Composition per liter:

Yeast extract ................................................................................. 2.0g

Trypticase peptone........................................................................ 2.0g

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

MgCl2·6H2O ................................................................................. 1.0g

CaCl2·2H2O .................................................................................. 0.4g

KH2PO4......................................................................................... 0.4g

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

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

Crotonate solution....................................................................10.0mL

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

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

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

Coenzyme M solution..............................................................10.0mL

pH 7.1 ± 0.2 at 25°C

Crotonate Solution:

Composition per 10.0mL:

Na-crotonate ................................................................................ 1.7g

Preparation of Crotonate Solution: Add Na-crotonate to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Sparge with 100% N2. Filter sterilize.

Coenzyme M Solution:

Composition per 10.0mL:

Mercaptoethanesulfonic acid (coenzyme M) .............................. 0.5g

Preparation of Coenzyme M: Add coenzyme M to distilled/de

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

with 100% N2. Filter sterilize.

Bicarbonate Solution:

Composition per 10.0mL:

NaHCO3........................................................................................ 2.0g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

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

with 20% CO2 + 80% H2. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to room temperature.

Sulfide Solution:

Composition per 10.0mL:

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

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.

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.

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 thoroughly to dissolve.

Preparation of Medium: Add components, except bicarbonate, vi

tamin, crotonate, coenzyme M, and sulfide solutions, to distilled/de

ionized water and bring volume to 950.0mL. Gently heat and bring to

boiling. Boil for 1 min. Mix thoroughly. Cool to room temperature

while sparging with 80% N2 + 20% CO2. Dispense into culture vessels

under an atmosphere of 80% N2 + 20% CO2 (e.g., Balch tubes or serum

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

anoxically add bicarbonate, vitamin, crotonate, coenzyme M, and sul

fide solutions. Adjust pH to 7.1.

Use: For the cultivation of Smithella spp.

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Smithella Medium (DSMZ Medium 1030)