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Steroidobacter Medium with Testosterone (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 Testosterone

(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 1040.3mL:

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

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

Testosterone solution ...............................................................10.0mL

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

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

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

pH 7.2 ± 0.2 at 25°C

Testosterone Solution:

Composition per 10.0mL:

Testosterone .................................................................................. 0.2g

Acetone....................................................................................10.0mL

Preparation of Testosterone Solution: Add tesosterone to acetone

adn bring volume to 10.0mL. Mix thoroughly.

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 components to dis

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

Sparge with 20% CO2 + 80% H2. Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to room temperature.

Preparation of Medium: Dispense testosterone solution into an

aerobic culture vessels. Allow solvent to evaporate to dryness. Add re

maining components, except bicarbonate solution, vitamin solution,

trace elements solution SL-10, and selenite-tungstate solution, to dis

tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Dis

pense 10.0mL portions of the medium 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 temper

ature. For every 10.0mL of medium, add 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 testosterone.

Use: For the cultivation of Steroidobacter denitrificans.

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Steroidobacter Medium with Testosterone (DSMZ Medium 1116)