Desulfitobacterium Medium (DSMZ Medium 663)

[所属分类:培养基配方] [发布时间:2020-6-4] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Desulfitobacterium Medium
(DSMZ Medium 663)

培养基配方
Composition per liter:
KH2PO4 .....................................................................5.44g
Yeast extract................................................................1.0g
NH4Cl .......................................................................0.5g
MgCl2 ·2H2O ...............................................................0.18g
CaCl2 ·2H2O ..............................................................0.032g
Resazurin...................................................................0.5mg
Vitamin solution...........................................................20.0mL
Na-pyruvate solution ......................................................10.0mL
Na-thiosulfate solution....................................................10.0mL
Cysteine solution .........................................................10.0mL
Na2S·9H2O solution........................................................10.0mL
Trace elements solution ....................................................5.0mL
pH 7.5 ± 0.2 at 25°C
Na-pyruvate Solution:
Composition per 10.0mL:
Na-pyruvate ................................................................2.5g
Preparation of Na-pyruvate Solution: Add Na-pyruvate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Na-thiosulfate Solution:
Composition per 10.0mL:
Na2S2O3 ·5H2O..............................................................2.5g
Preparation of Na-thiosulfate Solution: Add Na2S2O3 ·5H2O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Filter sterilize.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O...................................................................0.4g
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.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O .......................................................0.4g
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.
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid....................................................12.8g
FeCl3 ·6H2O.............................................................1.35g
NaCl......................................................................1.0g
CoCl2 ·4H2O ............................................................0.24g
NiCl2 ·6H2O.............................................................0.12g
MnCl2 ·4H2O..............................................................0.1g
CaCl2 ·2H2O..............................................................0.1g
ZnCl2 ....................................................................0.1g
Na2SeO3 ·5H2O..........................................................0.026g
CuCl2 ·2H2O ...........................................................0.025g
Na2MoO4 ·4H2O .........................................................0.024g
H3BO3 ...................................................................0.01g
Preparation of  Trace Elements Solution : Add nitrilotriacetic acid
to 500.0mL of distilled/deionized water. Dissolve by adjusting pH to
6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly. Adjust pH to 6.8.
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 B 12 ...............................................................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 an
oxygen-free atmosphere of 100% N2 . Add components, except vitamin
solution, cysteine solution, Na-pyruvate solution, Na-thiosulfate solu-
tion, and Na2S·9H2O solution, to distilled/deionized water and bring
volume to 940.0mL. Mix thoroughly. Sparge with 100% N2 . Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically and an-
aerobically add 20.0mL sterile vitamin solution, 10.0mL of sterile
cysteine solution, 10.0mL sterile Na-pyruvate solution, 10.0mL sterile
Na-thiosulfate solution, and 10.0mL of sterile Na2S·9H2O solution.
Mix thoroughly. Adjust pH to 7.5. Aseptically and anaerobically dis-
tribute into sterile tubes or flasks.
Use:  For the cultivation of Desulfitobacterium dehalogenans.