Clostridium ljungdahlii Medium (DSMZ Medium 879)

[所属分类:培养基配方] [发布时间:2020-5-12] [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

Clostridium ljungdahlii Medium
(DSMZ Medium 879)

培养基配方
Composition per liter:
NH4Cl .....................................................................1.0g
Yeast extract..............................................................1.0g
NaCl.......................................................................0.8g
MgSO4 ·7H2O...............................................................0.2g
KCl........................................................................0.1g
KH2PO4 ....................................................................0.1g
CaCl2 ·2H2O .............................................................0.02g
Na2WO4 ·2H2O............................................................0.20mg
Fructose solution........................................................50.0mL
Trace elements solution..................................................10.0mL
Vitamin solution.........................................................10.0mL
NaHCO3  solution.........................................................10.0mL
L -Cysteine solution.....................................................10.0mL
Na2S·9H2O solution......................................................10.0mL
pH 5.9 ± 0.2 at 25°C
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 ..................................................................1.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Fructose Solution:
Composition per 50.0mL:
Fructose.................................................................5.0g
Preparation of Fructose Solution: Add fructose to distilled/de-
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O...............................................................0.3g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Sparge with N2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
L -Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O ...................................................0.3g
Preparation of  L -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:
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: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, fructose solution,  L -cysteine solution, Na2S·9H2O solution, vita-
min solution, and trace elements solution SL-10, to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Gently heat and
bring to boiling. Boil for 10 min. Cool to room temperature while sparg-
ing with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 50.0mL fructose solution,
10.0mL NaHCO3  solution, 10.0mL  L -cysteine solution, 10.0mL
Na2S·9H2O solution, 10.0mL vitamin solution, and 10.0mL trace ele-
ments solution. Mix thoroughly. Final pH is 5.9. Aseptically and anaer-
obically distribute into sterile tubes or bottles.
Use:  For the cultivation of Clostridium ljungdahlii.