PYG Medium with Volatile Fatty Acids

[所属分类:培养基配方] [发布时间:2021-7-29] [发布人:网站管理员2] [阅读次数:] [返回]

PYG Medium with Volatile Fatty Acids

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

Composition per 1003.3mL:

Yeast extract................................................................................10.0g
Beef extract...................................................................................5.0g
Glucose.........................................................................................5.0g
Trypticase™..................................................................................5.0g
L -Cysteine·HCl .............................................................................0.5g
(NH 4 ) 2 SO 4 ....................................................................................0.5g
Hemin ........................................................................................5.0mg
Resazurin...................................................................................1.0mg
Mineral solution.......................................................................40.0mL
Fatty acid mixture......................................................................3.1mL
NaOH (1N solution)...................................................................0.5mL
Vitamin K 1 solution...................................................................0.2mL
pH 6.9 ± 0.2 at 25°C
Mineral Solution:
Composition per liter:
NaHCO 3 ......................................................................................10.0g
NaCl..............................................................................................2.0g
K 2 HPO 4 .........................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
MgSO 4 ·7H 2 O..............................................................................0.48g
CaCl 2 ·2H 2 O ..................................................................................0.3g
Preparation of Mineral Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Fatty Acid Mixture:
Composition per 31.0mL:
Acetic acid...............................................................................17.0mL
Propionic acid............................................................................6.0mL
n-Butyric acid............................................................................4.0mL
DL -2-Methylbutyric acid............................................................1.0mL
iso-Butyric acid..........................................................................1.0mL
iso-Valeric acid..........................................................................1.0mL
n-Valeric acid.............................................................................1.0mL
Preparation of Fatty Acid Mixture : Combine components.
Vitamin K 1 Solution:
Composition per 10.0mL:
Vitamin K 1 ..................................................................................0.05g
Ethanol (95% solution)............................................................10.0mL
Preparation of Vitamin K 1 Solution : Add vitamin K 1 to 95% eth-
anol and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Store in a brown glass bottle at 4°C.
Preparation of Medium: Dissolve hemin in 0.5mL of 1N NaOH.
Add remaining components, except  L -cysteine·HCl, and bring volume
to 1.0L with distilled/deionized water. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 5 min. Cool to room temperature
while sparging with 100% CO 2 . Add  L -cysteine·HCl. Mix thoroughly.
Adjust pH to 6.0 using 8N NaOH while continuing to sparge with
100% CO 2 . After pH has been attained, sparge with 100% N 2 . Anaer-
obically distribute into tubes or flasks. Autoclave for 15 min at 15 psi
pressure–121°C.
Use: For the cultivation and maintenance of Acetivibrio ethanolgign-
ens and Bacteroides xylanolyticus.