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Sludge Medium for Methanobacteria

[所属分类:培养基配方] [发布时间:2021-8-17] [发布人:网站管理员2] [阅读次数:] [返回]

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

Composition per liter:

NaHCO3........................................................................................ 4.0g

Sodium formate ............................................................................ 2.0g

Sodium acetate.............................................................................. 1.0g

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

L-Cysteine·HCl·H2O ..................................................................... 0.5g

KH2PO4......................................................................................... 0.5g

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

MgSO4·7H2O................................................................................ 0.4g

NaCl.............................................................................................. 0.4g

NH4Cl ........................................................................................... 0.4g

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

FeSO4·7H2O............................................................................... 2.0mg

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

Sludge fluid..............................................................................50.0mL

Fatty acid mixture ....................................................................20.0mL

Trace elements sollution SL-6 ...................................................1.0mL

pH 6.7-7.0 at 25°C

Sludge Fluid:

Composition per 100.0mL:

Yeast extract.................................................................................. 0.4g

Sludge ....................................................................................100.0mL

Preparation of Sludge Fluid: Add yeast extract to a concentration

of 0.4% to sludge taken from an anaerobic digester. Gas with 100% N2

for 5 min. Incubate at 37°C for 24 hr. Centrifuge at 13,000 × g. Remove

the supernatant fluid. Gas with 100% N2 for 5 min. Autoclave for 15

min at 15 psi pressure–121°C. Store at 25°C protected from light.

Fatty Acid Mixture:

Composition per 20.0mL:

α-Methylbutyric acid .................................................................... 0.5g

Isobutyric acid............................................................................... 0.5g

Isovaleric acid ............................................................................... 0.5g

Valeric acid ................................................................................... 0.5g

Preparation of Fatty Acid Mixture: Add components to distilled/

deionized water and bring volume to 20.0mL. Mix thoroughly. Adjust

pH to 7.5 with concentrated NaOH.

Trace Elements Solution SL-6:

Composition per liter:

H3BO3 ........................................................................................... 0.3g

CoCl2·6H2O .................................................................................. 0.2g

ZnSO4·7H2O ................................................................................. 0.1g

MnCl2·4H2O................................................................................ 0.03g

Na2MoO4·H2O ............................................................................ 0.03g

NiCl2·6H2O ................................................................................. 0.02g

CuCl2·2H2O................................................................................. 0.01g

Preparation of Trace Elements Solution SL-6: Add compo

nents to distilled/deionized water and bring volume to 1.0L. Mix thor

oughly. Adjust pH to 3.4.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2. Add components to distilled/deionized water and bring

volume to 1.0L. Mix thoroughly. Adjust pH to 6.7–7.0. Distribute an

aerobically into tubes or bottles with aluminum seals. Autoclave for 15

min at 15 psi pressure–121°C with fast exhaust.

Use: For the cultivation of Methanobacterium uliginosum and Metha

nobrevibacter ruminantium.

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Sludge Medium for Methanobacteria