Caldisphaera Medium (DSMZ Medium 991)

[所属分类:培养基配方] [发布时间:2020-3-26] [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

Caldisphaera Medium
(DSMZ Medium 991)

培养基配方
Composition per liter:
Sulfur, powder ..............................................10.0g
MnCl2 ·4H2O .................................................1.8g
(NH4)2SO4 ....................................................1.3g
KH2PO4 ......................................................0.28g
MgSO4 ·7H2O.................................................0.25g
CaCl2 ·2H2O ................................................0.07g
FeCl3 ·6H2O.................................................0.02g
Na2B4O7 ·10H2O..............................................4.5mg
Resazurin....................................................1.0mg
ZnSO4 ·7H2O................................................0.22mg
CuCl2 ·2H2O................................................0.05mg
Na2MoO4 ·4H2O..............................................0.03mg
VOSO4 ·2H2O................................................0.03mg
Na3 - citrate·2H2O.........................................0.03mg
CoSO4 ......................................................0.01mg
Vitamin solution............................................10.0mL
Na2S·9H2O solution.........................................10.0mL
Yeast extract solution.......................................5.0mL
pH 4.3 ± 0.2 at 25°C
Yeast Extract Solution:
Composition per 5.0mL:
Yeast extract................................................0.5g
Preparation of Yeast Extract Solution : Add yeast extract to dis-
tilled/deionized water and bring volume to 5.0mL. Mix thoroughly.
Sparge under 100% N2 gas for 3 min. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N2 gas.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O.....................................................0.5g
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.
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: Add components, except vitamin solution,
yeast extract solution, sulfur, and Na2S·9H2O solution, to distilled/deion-
ized water and bring volume to 975.0mL. Mix thoroughly. Gently heat
and bring to boiling. Boil for 3 min. Cool to room temperature under
80% N2 + 20% CO2 . Adjust pH to 3.5 with 10NH2SO4 . Dispense under
same gas atmosphere in suitable culture vessels (e.g., 20.0mL of the
medium in 120 mL serum bottles). Autoclave for 15 min at 15 psi pres-
sure–121°C. Steam sulfur for 3 hr on each of 3 successive days. Asep-
tically mix the sterilized sulfur with the medium and add vitamins and
yeast extract from sterile, anaerobic stock solutions. Prior to inocula-
tion change atmosphere to 80% H2 + 20% CO2 . Aseptically and anox-
ically add Na2S·9H2O. Adjust pH to 4.0–4.5 if necessary. After
inoculation pressurize vials to 1 bar overpressure with 80% H2 + 20%
CO2 gas mixture.
Use:  For the cultivation of Caldisphaera spp.