Deferribacter Medium (DSMZ Medium 935)

[所属分类:培养基配方] [发布时间:2020-6-2] [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

Deferribacter Medium
(DSMZ Medium 935)

培养基配方
Composition per liter:
NaCl...........................................................................25.0g
Sulfur, powdered...............................................................10.0g
Na-acetate......................................................................2.0g
KNO3 ...........................................................................0.5g
NH4Cl .........................................................................0.33g
KCl............................................................................0.33g
CaCl2 ·2H2O ..................................................................0.33g
MgCl2 ·6H2O ..................................................................0.33g
KH2PO4 ........................................................................0.33g
Yeast extract..................................................................0.15g
NaHCO3 solution...............................................................10.0mL
Trace elements solution.......................................................10.0mL
Vitamin solution..............................................................10.0mL
pH 7.0 ± 0.2 at 25°C
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 ........................................................................0.3g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared
freshly.
Trace Elements Solution:
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:  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.
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,
NaHCO3 solution, and sulfur, to 980.0mL distilled/deionized water.
Gently heat and bring to boiling. Boil for 3 min. Cool to room tempera-
ture while sparging with 100% N2 . Adjust pH to 7.0. Anaerobically un-
der 100% N2  distribute into tubes of bottles containing the sulfur (0.1g
sulfur per 10mL medium). Autoclave for 20 min at 110°C. Aseptically
and anaerobically add 10.0mL sterile vitamin solution and 10.0mL ster-
ile NaHCO3 solution. Mix thoroughly. The final pH should be 7.0.
Use: For the cultivation of Deferribacter desulfuricans and Deferri-
bacter thermophilus.