Anaerobic Acetoin Medium

[所属分类:培养基配方] [发布时间:2019-10-11] [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

Anaerobic Acetoin Medium
Composition per 1006.0mL:
Solution A..............................................................................916.0mL
Solution B................................................................................70.0mL
Solution C................................................................................10.0mL
Solution D................................................................................10.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per 916.0mL:
Acetoin..........................................................................................1.5g
Pancreatic digest of casein............................................................1.0g
Resazurin...................................................................................1.0mg
Mineral solution.......................................................................50.0mL
Rumen fluid, clarified..............................................................50.0mL
Vitamin solution.........................................................................5.0mL
Trace elements solution SL-10..................................................1.0mL
Mineral Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................12.5g
NaCl..............................................................................................1.0g
FeCl 3 ·4H 2 O...................................................................................0.2g
MnCl 2 ·4H 2 O.................................................................................0.1g
CaCl 2 ·2H 2 O..................................................................................0.1g
ZnCl 2 ............................................................................................0.1g
CuCl 2 ..........................................................................................0.02g
Na 2 SeO 3 ......................................................................................0.02g
CoCl 2 ·6H 2 O..............................................................................0.017g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Mineral Solution : Add nitrilotriacetic acid to
500.0mL of distilled/deionized water. Adjust 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..........................................................................6.2mg
Nicotinic acid.............................................................................2.5mg
Thiamine·HCl..........................................................................1.25mg
p-Aminobenzoic acid...............................................................1.25mg
Pantothenic acid.......................................................................0.62mg
Biotin.......................................................................................0.25mg
Preparation of Vitamin Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O.......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 916.0mL. Adjust pH to 7.2. Gently heat and
bring to boiling. Continue boiling for a few minutes. Allow to cool to
room temperature under 80% N 2 + 20% CO 2 . Distribute into bottles un-
der 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 70.0mL:
NaHCO 3 ........................................................................................3.5g
Preparation of Solution B:  Add NaHCO 3 to distilled/deionized
water and bring volume to 70.0mL. Mix thoroughly. Filter sterilize.
Sparge with 80% N 2 + 20% CO 2 for 15 min.
Solution C:
Composition per 10.0mL:
L -Cysteine·HCl..............................................................................0.3g
Preparation of Solution C: Add  L -cysteine·HCl to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N 2 for 3–4 min. Autoclave under 100% N 2  for 15 min at 15 psi
pressure–121°C.
Solution D:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Solution D: Add Na 2 S·9H 2 O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N 2 for 3–4 min. Autoclave under 100% N 2  for 15 min at 15 psi pres-
sure–121°C.
Preparation of Medium: To 916.0mL of sterile solution A add
70.0mL of sterile solution B, 10.0mL of sterile solution C, and 10.0mL
of sterile solution D. Mix thoroughly.
Use:  For the cultivation of anaerobic bacteria that can metabolize ace-
toin.