BC Medium (Medium for Acetivibrio cellulolyticus)

[所属分类:培养基配方] [发布时间:2019-12-9] [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

BC Medium
(Medium for Acetivibrio cellulolyticus)

培养基配方
Composition per liter:
Cellulose powder .........................................3.0g
NaHCO 3 ..................................................2.0g
Mineral solution 1......................................75.0mL
Mineral solution 2......................................75.0mL
Cysteine-sulfide reducing solution .....................12.8mL
FeSO4 ·7H2O solution...................................10.0mL
Vitamin mixture.........................................10.0mL
Wolfe’s mineral solution...............................10.0mL
Resazurin (0.1% solution)................................1.0mL
pH 7.6 ± 0.2 at 25°C
Caution: This medium contains sodium sulfide and may produce tox-
ic H2S gas. Prepare in a chemical fume hood.
Mineral Solution 1:
Composition per liter:
K2HPO4 ......................................................3.9g
Preparation of Mineral Solution 1: Add K 2 HPO 4 to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly.
Mineral Solution 2:
Composition per liter:
NH4Cl ...................................................12.0g
Na2SO4 ...................................................2.5g
KH2PO4 ...................................................2.4g
MgSO4 ·7H2O..............................................1.2g
CaCl2 ·2H2O .............................................0.8g
Preparation of Mineral Solution 2: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
FeSO4 ·7H2O Solution:
Composition per 100.0mL:
FeSO4 ·7H2O...............................................0.2g
Preparation of FeSO4 ·7H2O Solution: Dissolve FeSO4 ·7H2O in
100.0mL of distilled/deionized water. Add three drops of concentrated
HCl. Mix thoroughly.
Vitamin Mixture:
Composition per liter:
Pyridoxine·HCl...........................................10.0mg
Thiamine·HCl..............................................5.0mg
Cyanocobalamin.............................................5.0mg
Lipoic acid (thioctic acid)................................5.0mg
Biotin.....................................................2.0mg
p-Aminobenzoic acid........................................0.5mg
Preparation of Vitamin Mixture: Add components to distilled/
deionized water and bring volume to 1.0L. Store below −20°C.
Wolfe’s Mineral Solution:
Composition per liter
MgSO4 ·7H2O.................................................3.0g
Nitriloacetic acid...........................................1.5g
MnSO4 ·H2O..................................................0.5g
NaCl.........................................................1.0g
FeSO4 ·7H2O.................................................0.1g
CoCl2 ·6H2O.................................................0.1g
CaCl2 .......................................................0.1g
ZnSO4 ·7H2O.................................................0.1g
CuSO4 ·5H2O................................................0.01g
AlK(SO4)2 ·12H2O...........................................0.01g
H3BO3 ......................................................0.01g
Na2MoO4 ·2H2O..............................................0.01g
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water and adjust to pH 6.5 with
KOH to dissolve. Bring volume to 1.0L with distilled/deionized water.
Add remaining components one at a time.
Cysteine-Sulfide Reducing Solution:
Composition per 200.0mL:
L- Cysteine·HCl·H2O .........................................2.5g
Na2S·9H2O.....................................................2.5g
Preparation of Cysteine-Sulfide Reducing Solution: Add  L-
cysteine·HCl·H 2 O to 50.0mL of distilled/deionized water. Quickly ad-
just pH to 10 with fresh 3N NaOH and flush under 100% N2 . Add
Na2S·9H2O. Bring volume to 200.0mL with distilled/deionized water.
Boil under 100% N 2 . Transfer anaerobically to tubes or flasks and stop-
per. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add cellulose and NaHCO3 to distilled/de-
ionized water and bring volume to 800.0mL. Add all other components ex-
cept cysteine-sulfide reducing solution. Heat and boil under 90% N2 +
10% CO2 . Cool and continue flushing under 90% N2 + 10% CO2 . The pH
should be 7.6 at room temperature; do not adjust. Add 8.0mL of cysteine-
sulfide reducing solution. Add 4.8mL more of cysteine-sulfide reducing
solution. Distribute anaerobically into tubes in 7.0mL volumes and cap.
Use:  For the cultivation and maintenance of Acetivibrio cellulolyticus,
Acetivibrio cellulosolvens, Bacteroides cellulosolvens, and other cellu-
lose-degrading microorganisms.