Propionivibrio/Acetivibrio/Formivibrio Medium

[所属分类:培养基配方] [发布时间:2021-7-14] [发布人:网站管理员2] [阅读次数:] [返回]

Propionivibrio/Acetivibrio/Formivibrio Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1012.0mL:

Solution A..............................................................................950.0mL
Solution E ................................................................................30.0mL
Solution D (Vitamin solution)..................................................10.0mL
Solution F.................................................................................10.0mL
Solution G................................................................................10.0mL
Solution B (Trace elements solution SL-10)..............................1.0mL
Solution C (Selenite-tungstate solution)....................................1.0mL
pH 7.7–7.9 at 25°C
Solution A:
Composition per 950.0mL:
KH 2 PO 4 .........................................................................................1.4g
NH 4 Cl ...........................................................................................0.5g
MgCl 2 ·6H 2 O..................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
Yeast extract.............................................................................50.0mg
Resazurin ...................................................................................1.0mg
Preparation of Solution A : Add components to distilled/deionized
water and bring volume to 950.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution B (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 Solution B (Trace Elements Solution SL-10):
Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min
at 15 psi pressure–121°C.
Solution C (Selenite-Tungstate Solution):
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Solution C (Selenite-Tungstate Solution):
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution D (Vitamin Solution):
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Solution D (Vitamin Solution):  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Solution E:
Composition per 30.0mL:
NaHCO 3 ........................................................................................1.5g
Preparation of Solution E:  Add NaHCO 3 to distilled/deionized
water and bring volume to 30.0mL. Mix thoroughly. Filter sterilize.
Sparge with 100% N 2 .
Solution F:
Composition per 10.0mL:
Cinnamic acid...............................................................................1.6g
NaOH (1N solution)................................................................ variable
Preparation of Solution F:  Add cinnamic acid to distilled/deion-
ized water and bring volume to 8.0mL. Mix thoroughly. Add sufficient
quantity of 1N NaOH solution to bring pH to 7.8. Sparge with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution G:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.25g
Preparation of Solution G:  Add Na 2 S·9H 2 O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically under
100% N 2 combine 950.0mL of sterile solution A with 1.0mL of sterile
solution B, 1.0mL of sterile solution C, 10.0mL of sterile solution D,
30.0mL of sterile solution E, 10.0mL of sterile solution F, and 10.0mL
of sterile solution G, in that order. Mix thoroughly. Final pH should be
7.7–7.9. If necessary, add about 15.0mL of sterile anaerobic 5%
Na 2 CO 3  solution to 1.0L of medium to adjust pH. Aseptically and an-
aerobically distribute into sterile tubes or flasks under 100% N 2 .
Use:  For the cultivation and maintenance of Acetivibrio multivorans .