165 .
ACETIVIBRIO CELLULOLYTICUS MEDIUM
What is this page?
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.2 - 7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Mineral solution 1 | 75.00 | ml | ||
|
|
Mineral solution 2 | 75.00 | ml | ||
|
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
|
FeSO4 x 7 H2O solution (0.1% w/v) | 20.00 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Na2CO3 | 1.50 | g | ||
|
|
Cellobiose | 3.00 | g | ||
|
|
Cellulose
(MN300) optional |
3.00 | g | ||
|
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.25 | g | ||
|
|
Na2S x 9 H2O | 0.25 | g | ||
|
|
Distilled water | 820.00 | ml | ||
| 1 Dissolve ingredients except carbonate, cellobiose, vitamins, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes and autoclave. Add cellobiose, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Cellobiose and vitamins have to be sterilized by filtration. Prior to use adjust pH of complete medium to 7.2 - 7.4. Cellulose (MN 300, Whatman CF II, Kleenex tissue paper, or HCl treated cotton) can be used as an alternative to cellobiose. | |||||
Mineral solution 2 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 2.40 | g |
|
|
(NH4)2SO4 | 6.00 | g |
|
|
NaCl | 0.59 | g |
|
|
MgSO4 x 7 H2O | 1.20 | g |
|
|
CaCl2 x 2 H2O | 0.72 | g |
|
|
Distilled water | 1000.00 | ml |
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
MnSO4 x H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.18 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.18 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.30 | mg | ||
|
|
Na2WO4 x 2 H2O | 0.40 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||
FeSO4 x 7 H2O solution (0.1% w/v) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
FeSO4 x 7 H2O | 1 | g | ||
|
|
H2SO4
(0.1 N) |
1000 | ml | ||
| 1 The ferrous sulfate solution is not stable and should be freshly prepared. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
|
Thiamine HCl | 50 | mg |
|
|
Riboflavin | 50 | mg |
|
|
Nicotinic acid | 50 | mg |
|
|
Calcium D-(+)-pantothenate | 50 | mg |
|
|
Vitamin B12 | 1 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |