Medium for Roseospira (DSMZ Medium 998)

[所属分类:培养基配方] [发布时间:2021-5-20] [发布人:网站管理员2] [阅读次数:] [返回]

Medium for Roseospira
(DSMZ Medium 998)

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

Composition per liter:

NaCl............................................................................................20.0g
MgCl 2 ·6H 2 O..................................................................................1.0g
MgSO 4 · 7H 2 O..............................................................................0.25g
NH 4 Cl ...........................................................................................0.5g
Yeast extract..................................................................................0.5g
KH 2 PO 4 .........................................................................................0.3g
CaCl 2 ·2H 2 O.................................................................................0.05g
NaHCO 3  soltuion .....................................................................10.0mL
Acetate solution .......................................................................10.0mL
Succinate solution....................................................................10.0mL
Trace elements solution SL-12 ..................................................1.0mL
Vitamin V7 solution...................................................................1.0mL
pH 6.9 ± 0.2 at 25°C
Acetate Solution:
Composition per 10.0mL:
Sodium acetate............................................................................0.41g
Preparation of Acetate Solution:  Add sodium acetate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO 2 + 80% N 2 . Filter sterilize.
Succinate Solution:
Composition per 10.0mL:
Sodium succinate........................................................................0.85g
Preparation of Succinate Solution:  Add sodium succinate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 20% CO 2 + 80% N 2 . Filter sterilize.
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO 2 + 80% N 2 . Filter sterilize.
Vitamin Solution V7:
Composition per liter:
Pyridoxine-HCl........................................................................50.0mg
Nicotinic acid...........................................................................20.0mg
Vitamin B 12 ..............................................................................20.0mg
Thiamine-HCl·2H 2 O................................................................10.0mg
p-Aminobenzoic acid...............................................................10.0mg
D -Ca-pantothenate .....................................................................5.0mg
Biotin.........................................................................................2.0mg
Preparation of Vitamin Solution V7:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
Trace Elements Solution SL-12:
Composition per liter:
FeSO 4 ·7H 2 O..................................................................................1.1g
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O................................................................................0.19g
MnCl 2 ·2H 2 O ...............................................................................0.05g
ZnCl 2 ........................................................................................42.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
Na 2 MoO 4 ·4H 2 O.......................................................................18.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
Preparation of Trace Elements Solution Sl-12:  Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Filter sterilize.
Preparation of Medium: Add components, except bicarbonate, vi-
tamin, acetate, and succinate solutions, to distilled/deionized water and
bring volume to 970.0mL. Mix thoroughly. Gently heat while stirring
and bring to boiling. Boil for 1 min. Cool to room temperature while
sparging with 90% N 2 + 10% CO 2 gas. Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically add bicarbonate and vitamin solutions.
Mix thoroughly. Adjust pH to 6.9. Distribute into sterile 50mL screw-
capped bottles. Add the organic acetate and succinate substrates.
Use: For the cultivation of Roseospira spp.