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Sphaerotilus natans Medium (LMG Medium 33)

[所属分类:培养基配方] [发布时间:2021-8-20] [发布人:网站管理员2] [阅读次数:] [返回]

Sphaerotilus natans Medium

(LMG Medium 33)

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

Peptone.......................................................................................... 5.0g

Casitone ........................................................................................ 5.0g

Glucose ......................................................................................... 5.0g

(NH4)2SO4..................................................................................... 0.5g

L-Cysteine·HCl.............................................................................. 0.5g

Resazurin ...................................................................................1.0mg

Mineral solution.......................................................................40.0mL

Fatty acid mixture ......................................................................3.1mL

Hemin solution...........................................................................0.5mL

Vitamin K1 .................................................................................0.2mL

pH 6.9 ± 0.2 at 25°C

Mineral Solution:

Composition per liter:

NaHCO3...................................................................................... 10.0g

NaCl.............................................................................................. 2.0g

K2HPO4......................................................................................... 1.0g

KH2PO4......................................................................................... 1.0g

MgSO4·7H2O.............................................................................. 0.48g

CaCl2·2H2O .................................................................................. 0.3g

Preparation of Mineral Solution: Add components to distilled/

deionized water and bring volume to 1.0L. Mix thoroughly.

Fatty Acid Mixture:

Composition per 31.0mL:

Acetic acid ...............................................................................17.0mL

Propionic acid ............................................................................6.0mL

n-Butyric acid ............................................................................4.0mL

n-Valeric acid.............................................................................1.0mL

iso-Valeric acid ..........................................................................1.0mL

iso-Butyric acid..........................................................................1.0mL

DL-2-Methylbutyric acid............................................................1.0mL

Preparation of Fatty Acid Mixture: Combine components. Mix

thoroughly. Adjust pH to 7.5 with concentrated NaOH.

Hemin Solution:

Composition per 1.0mL:

Hemin ........................................................................................ 5.0mg

NaOH (1N solution)...................................................................1.0mL

Preparation of Hemin Solution: Add hemin to 1.0mL of NaOH so

lution. Mix thoroughly.

Preparation of Medium: Add components, except L-cysteine·HCl,

hemin solution, and fatty acid mixture, to distilled/deionized water and

bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boil

ing. Continue boiling for 5 min. Cool to room temperature while sparg

ing with 100% CO2. Add L-cysteine·HCl, hemin solution, and fatty

acid mixture. Adjust pH to 6.9 with 8N NaOH while continuing to

sparge with 100% CO2. After pH has been reached, sparge with 100%

N2. Anaerobically distribute into tubes or flasks. Autoclave for 15 min

at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Sphaerotilus natans.

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Sphaerotilus natans Medium (LMG Medium 33)