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Sulfurospirillum II Medium (DSMZ Medium 771)

[所属分类:培养基配方] [发布时间:2021-8-31] [发布人:网站管理员2] [阅读次数:] [返回]

Sulfurospirillum II Medium

(DSMZ Medium 771)

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

Composition per 1080.0mL:

Yeast extract.................................................................................. 1.0g

NaCl .......................................................................................460.0mg

K2HPO4..................................................................................225.0mg

KH2PO4..................................................................................225.0mg

(NH4)2SO4..............................................................................225.0mg

MgSO4·7H2O ......................................................................... 117.0mg

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

NaHCO3 solution .....................................................................30.0mL

NaNO3 solution ........................................................................10.0mL

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

Na-lactate solution ...................................................................10.0mL

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

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

Trace elements solution SL-10 ..................................................1.0mL

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

pH 7.3 ± 0.2 at 25°C

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.

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.

Na-lactate Solution:

Composition per 10.0mL:

Na-lactate.................................................................................... 2.25g

Preparation of Na-lactate Solution: Add Na-lactate to distilled/

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

with 100% N2. Filter sterilize.

NaNO3 Solution:

Composition per 10.0mL:

NaNO3 .......................................................................................... 1.7g

Preparation of NaNO3 Solution: Add NaNO3 to distilled/deion

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

80% N2 + 20% CO2. Filter sterilize.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.1g

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.

NaHCO3 Solution:

Composition per 30.0mL:

NaHCO3........................................................................................ 4.2g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Filter sterilize.

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 Trace Elements Solution SL-10: Add FeCl2·4H2O

to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized

water and bring volume to 1.0L. Add remaining components. Mix thor

oughly. Sparge with 80% N2 + 20% CO2.

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

solution, cysteine solution, NaNO3 solution, Na-lactate solution, and

vitamin solution, to distilled/deionized water and bring volume to

1.0mL. Mix thoroughly. Sparge with 80% N2 + 20% CO2. Adjust pH to

7.3. Dispense either 10.0mL aliquots into 15mL Hungate tubes or

50.0mL aliquots into 100mL Hungate bottles. Autoclave for 15 min at

15 psi pressure–121°C. Aseptically and anaerobically inject from sterile

stock solutions NaHCO3 solution, Na2S·9H2O solution, cysteine solu

tion, vitamin solution, sodium nitrate solution, and sodium lactate so

lution. Final pH should be 7.3.

Use: For the cultivation of Sulfurospirillum arsenophilum (Geospiril

lum sp.) and Sulfurospirillum barnesii.

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Sulfurospirillum II Medium (DSMZ Medium 771)