Peptone Yeast Extract Glucose Medium, Modified (MPYG Medium)

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

Peptone Yeast Extract Glucose Medium, Modified
(MPYG Medium)

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

Composition per 950.0mL:

Peptone .......................................................................................10.0g
Yeast extract................................................................................10.0g
Glucose.........................................................................................5.0g
L -Cysteine·HCl·H 2 O.....................................................................0.5g
(NH 4 ) 2 SO 4 ....................................................................................0.5g
Salt solution.............................................................................40.0mL
Vitamin K-heme solution.........................................................10.0mL
Resazurin (0.025% solution) .....................................................4.0mL
Volatile fatty acid solution.........................................................3.1mL
pH 7.0 ± 0.2 at 25°C
Salt Solution:
Composition per liter:
NaHCO 3 ......................................................................................10.0g
NaCl..............................................................................................2.0g
K 2 HPO 4 .........................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
CaCl 2 , anhydrous..........................................................................0.2g
MgSO 4 ..........................................................................................0.2g
Preparation of Salt Solution:  Add CaCl 2 and MgSO 4 to 300.0mL
of distilled/deionized water. Mix thoroughly until dissolved. Bring vol-
ume to 800.0mL with distilled/deionized water. Add remaining com-
ponents while stirring. Bring volume to 1.0L. Mix thoroughly. Store at
4°C.
Vitamin K-Heme Solution:
Composition per liter:
Part A.....................................................................................100.0mL
Part B.........................................................................................1.0mL
Preparation of Vitamin K-Heme Solution: Aseptically add
1.0mL of sterile part B to 100.0mL of cooled sterile part A. Mix thor-
oughly.
Part A:
Composition per 100.0mL:
Hemin ......................................................................................50.0mg
NaOH (1N solution)...................................................................1.0mL
Preparation of Part A: Add hemin to NaOH solution and bring
volume to 100.0mL with distilled/deionized water. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Part B:
Composition per 30.0mL:
Menadione (vitamin K 3 )........................................................100.0mg
Ethanol (95% solution)............................................................30.0mL
Preparation of Part B:  Add menadione to ethanol. Mix thoroughly.
Filter sterilize.
Volatile Fatty Acid Solution:
Composition per 31.0mL:
Acetic acid...............................................................................17.0mL
Propionic acid............................................................................6.0mL
n-Butyric acid ............................................................................4.0mL
n-Valeric acid.............................................................................1.0mL
Isovaleric acid............................................................................1.0mL
Isobutyric acid............................................................................1.0mL
DL- α-Methyl butyric acid...........................................................1.0mL
Preparation of Volatile Fatty Acid Solution: Combine compo-
nents. Mix thoroughly.
Preparation of Medium: Add components—except vitamin K-
heme solution,  L -cysteine·HCl·H 2 O, and volatile fatty acid solution—
to distilled/deionized water and bring volume to 936.9mL. Gently heat
and bring to boiling under 97% N 2 + 3% H 2 . Continue boiling until re-
sazurin turns colorless, indicating reduction. Cool to 45°–50°C. Add
vitamin K-heme solution,  L -cysteine·HCl·H 2 O, and volatile fatty acid
solution. Adjust pH to 7.0. Distribute into tubes under 97% N 2 + 3%
H 2 . Cap with rubber stoppers. Place tubes in a press. Autoclave for 15
min at 15 psi pressure–121°C with fast exhaust.
Use:  For the cultivation and maintenance of Acetivibrio ethanolgign-
ens, Butyrivibrio fibrisolvens, Lachnospira multipara, and Succinivi-
brio dextrinosolvens.