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Steroidobacter Medium with Heptanoate (DSMZ Medium 1116)

[所属分类:培养基配方] [发布时间:2021-8-30] [发布人:网站管理员2] [阅读次数:] [返回]

Steroidobacter Medium with Heptanoate

(DSMZ Medium 1116)

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

Composition per 1090.3mL:

NaCl ............................................................................................. 1.0g

KCl ............................................................................................... 0.5g

NaNO3......................................................................................... 0.42g

MgCl2·6H2O.................................................................................. 0.4g

NH4Cl ........................................................................................ 0.25g

KH2PO4 ........................................................................................ 0.2g

CaCl2·2H2O................................................................................. 0.15g

Na2SO4........................................................................................ 0.07g

Heptanoate solution .................................................................50.0mL

Bicarbonate solution ................................................................30.0mL

Trace elements soltuion SL-10 ..................................................0.1mL

Selenite-tungstate solution.........................................................0.1mL

Vitamin solution.........................................................................0.1mL

pH 7.2 ± 0.2 at 25°C

Heptanoate Solution:

Composition per 100.0mL:

Heptanoate .................................................................................. 0.85g

Preparation of Heptanoate Solution: Add components to dis

tilled/deionized water and bring volume to 100.0mL. Mix thoroughly

for several hours. Filter sterilize.

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 970.0mL. Mix thor

oughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

Trace Elements Solution SL-10:

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 SL-10: Add nitrilotri

acetic acid to 500.0mL of distilled/deionized water. Dissolve by adjust

ing pH to 6.5 with KOH. Add remaining components. Add distilled/

deionized water to 1.0L. Mix thoroughly. Adjust pH to 7.0.

Vitamin Solution:

Composition per liter:

Vitamin B12 .............................................................................. 50.0mg

Thiamine-HCl·2H2O................................................................ 50.0mg

Riboflavin ................................................................................ 50.0mg

D-Ca-pantothenate ................................................................... 50.0mg

p-Aminobenzoic acid............................................................... 50.0mg

Lipoic acid ............................................................................... 50.0mg

Nicotinic acid........................................................................... 25.0mg

Nicotine amide......................................................................... 25.0mg

Biotin ....................................................................................... 20.0mg

Folic acid ................................................................................. 20.0mg

Pyridoxine-HCl........................................................................ 10.0mg

Preparation of Vitamin Solution: Add components to distilled/

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

hours. Filter sterilize.

Bicarbonate Solution:

Composition per 100.0mL:

NaHCO3........................................................................................ 8.4g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge

with 20% CO2 + 80% N2. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to room temperature.

Preparation of Medium: Add components, except heptanoate so

lution, bicarbonate solution, vitamin solution, trace elements solution,

and selenite-tungstate solution, to distilled/deionized water and bring

volume to 1.0L. Mix thoroughly. Dispense 10.0mL portions of the me

dium to the culture vessels. Sparge with a gas mixture of 80% N2 +

20% CO2. Close the culture vessels. Autoclave for 15 min at 15 psi

pressure–121°C. Cool to room temperature. For every 10.0mL of me

dium, add 0.5mL heptanoate solution, 0.3mL bicarbonate solution,

0.001mL trace elements solution SL-10, 0.001mL selenite-tungstate

solution, and 0.001mL vitamin solution. Adjust pH to 7.2. Treat the

vessels in an ultrasonic bath to detach and suspend the heptanoate.

Use: For the cultivation of Steroidobacter denitrificans.

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Steroidobacter Medium with Heptanoate (DSMZ Medium 1116)