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Thermosyntropha Medium (DSMZ Medium 731)

[所属分类:培养基配方] [发布时间:2021-9-8] [发布人:网站管理员2] [阅读次数:] [返回]

Thermosyntropha Medium

(DSMZ Medium 731)

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................................................................................ 10.0g

Na2CO3 ......................................................................................... 3.0g

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

NaCl.............................................................................................. 0.5g

K2HPO4......................................................................................... 0.3g

KCl............................................................................................... 0.3g

MgCl2·6H2O ................................................................................. 0.3g

CaCl2·2H2O ................................................................................ 0.05g

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

Vitamin solution.......................................................................40.0mL

Trace elements solution ...........................................................10.0mL

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

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

NaHCO3 solution .....................................................................10.0mL

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

pH 8.5 ± 0.2 at 25°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.

NaHCO3 Solution:

Composition per 10.0mL:

NaHCO3........................................................................................ 3.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

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

for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared

freshly.

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine-HCl·H2O................................................................... 0.15g

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:

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.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

tilled/deionized water and bring volume to 10.0mL. Sparge with N2.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an

aerobically.

Preparation of Medium: Prepare and dispense medium under 100%

N2. Add components, except NaHCO3 solution, Na2S·9H2O solution,

cysteine solution, vitamin solution, selenite-tungstate solution, and

trace elements solution, to distilled/deionized water and bring volume to

919.0mL. Mix thoroughly. Sparge with 100% N2. Adjust pH to 8.5 with

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

aerobically add 10.0mL NaHCO3 solution, 10.0mL Na2S·9H2O solu

tion, 10.0mL cysteine solution, 40.0mL vitamin solution, 1.0mL

selenite-tungstate solution, and 10.0mL trace elements solution. Mix

thoroughly. Aseptically and anaerobically distribute into sterile tubes or

bottles.

Use: For the cultivation of Thermosyntropha lipolytica.

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Thermosyntropha Medium (DSMZ Medium 731)